STUDIES ON T H E BIOCHEMICAL R E S P O N S E S OF SOME SELECTED
PERENNIALS TO COAL-SMOKE POLLUTION
Ph. D. THESIS IN
BOTANY
SAHEED. S.
DEPARTMENT OF BOTANY ALIGARH MUSLIM UNIVERSITY
ALIGARH (INDIA) 1994
•ZAD
r
T4700
IN THE NAME OF ALLAH, THE MOST GRACIOUS, MOST MERCIFUL
Mischief has appeared on land and sea because of (the meed) that the hands of men have earned. That (God) may give them a taste of some of their deeds : inorder that they may turn back.
[Holy Quran, 30:19]
Nay, but you are people who have wasted their own selves. [Holy Quran, 36:19]
^ ^ ^ ^ ^ ^ ^ ^
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(iyE'DiavPE(D TO 'BCE
fM^EMOfRif 0 5 f W i T "
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ALIGARH MUSLIM UNIVERSITY DEPARTMENT OF BOTANY, ALIGARH-202 002, INDIA Tel. (0571) 25676
A. K. M. GHOUSE Ph.D., F.L.S.. FJ^M
ProfvMor 6 Chairman 0»terf
CERTIFICATE
This is to certify that the thesis entitled "Studies on the biochemical responses
of some selected perennials to coal-smoke pollution" embodies original and bonafide
work carried out under my supervision by Mr. Saheed, S. It may be submitted to the
Aligarh Muslim University, Aiigarh towards the fulfilment of requirements for the degree
of Doctor of Philosphy in Botany.
(A.K.M. Ghouse)
ACKNOWLEDGEMENTS
1 bow in reverence to Almtghly, Allah whose benign benediction gave me the
required zeal for the completion of this work.
Words are limited and the pages are not sufficient to express my sincere
thanks and debt to my venerable supervisor Prof. A.K.M. Ghouse, Ex-Chairman,
Department of Botany, Aligarh Muslim University, Aligarh, for his brilliant super-
vision, constant encouragement, invigorating suggetions and boundless generosity
throughout the period of this research study at AMU. It was only due to his immense
help, one of my long cherished ambitions has come to this shape, as a last man in the
queue under his expert guidance. His fruitful discussions and constructive criticism
have always been a source of great inspiration to me.
No less is my indebtedness to Prof. Wazahat Hussain, the present Chairman
of Department of Botany, AMU, for his encouragement by providing necessary
laboratory and library facilities throughout the present investigation.
I also feel indebted to Prof. Ziauddin Ahmad, Department of Botany, who
has always been my well wisher and encouraged me with his rich experience and
productive knowledge during the entire period of the present study.
Deep appreciation is extended to my senior laboratory colleagues, Dr.
Farooq A. Lone and Mrs. Azra Parveen, for their ungrudging help and constant
inspiration. A special credit deserves the former who always assisted me efficiently
in each and every stage of this work, and this work would not have been completed
without his limitless help.
A grateful acknowledgement is also extended to Drs. Farced A. Khan, P.R.
Khan and M.I.H. Khan, for their cherished co-operation in different ways.
J place on records my heartfelt thanks to Dr. Munawar Fazal, Dr. Imran
Khan, Dr. Hisamuddin, Dr. M.J. Pasha, Dr. KalintuUah and Dr. N.H. Shah, for
their critical views, total involvement and co-operation in bringing out this work.
1 acknowledge with immense pleasure from the core of my heart, the help received
from Mr. Abbas T.P., Mr. Kunhalavi, M. Mr. Subramanian, G.A. and Mr. Faizal,
A.U., at the time of dire need.
Many of myfriends deserve to be mentioned here, especially Mr. A bduraheem
K., Mr. Lateef, C.P.A., Dr. Sakeer Husain, Mr. Azees, TV., Mr. Abubakkar, K.K.,
Mr. Ashaq Raza,Mr. Shamsul Hayat, Mr. Moinuddin Khan and Mr. Majid Kazi,
their mere association, encouragement and accompaniment have helped me a lot.
Thanks are also due to Mr. Aslam Raza, Mr. S. Waheed andAsim Raza of
KLIC, Aligarh for the excellent laser printing and patience.
The financial assistance rendered by the University Grants Commission
(UGC), New Delhi in the form of Junior and Senior Research Fellowships is also
gratefully acknowledged.
Finally, it will be unfair on my part if I do not aknowledge my elder brothers
and sisters, who have been generous engough to inspire me to cross the horizon of
this too long and rather irksome academic career. They stood shoulder to shoulder
throughout my past and present. —
(SAHEED, S.) ' •
CONTENTS Page No,
INTRODUCTION 1 - 3
GEOGRAPHICAL SET-UP OF THE STUDY AREA 4 - 7
MATERIALS AND METHODS 8 - 2 0
OBSERVATIONS 21 - 4 7
Sulphur 2 1 - 2 3 Photosynthetic pigments 23-24 Carbohydrate 24 -27 Protein 27 -30 Nitrogen 3 0 - 3 3 Phosphorus 33-36 Potassium 36-39 Sodium 39-42 Calcium 42 -45 Ascorbic Acid 45 -46 Proline 46-47
DISCUSSION 4 8 - 8 9
Sulphur 48 -51 Chlorophyll Pigments 52 -55 Carotenoids 56-57 Carbohydrate 58-61 Protein" 62 -64 Nitrogen 65 -68 Phosphorus 69 -72 Potassium 73-76 Sodium 77-80 Calcium 8 1 - 8 3 Ascorbic Acid 8 4 - 8 6 Proline 87 -89
Page No.
COMPARATIVE PERFORMANCE OF 90 - 91 THE SELECTED SPECIES
CONCLUSION 92 - 93
SUMMARY 94 -100
REFERENCES 101 -125
APPENDIX I - II
INTRODUCTION
The coal based thermal power plants are the point sources of air pollution with
a more or less definite pattern of emission. The organic fuel burnt at thermal power
plants contains harmful impurities which are injected into the environment in gaseous
form (SOj, NOjj, CO, HC's and Fluoride etc.) and solid components of combustion
products (Fly ash, Bottom ash. Particulate matter etc.) which cause deleterious
effects on the whole complex of the hving system or the biosphere including the
vegetation, aquatic life of the region, animals and man. These thermal power plants
use thousands of tons of low quality, high ash content coal per day (Indian bituminous
coal contains 15 to 45 per cent ash against 3 to 5 per cent of in situ formed European
coal). A super thermal power plant using even normal or low sulphur coal will emit
about 100 tons of sulphur dioxide into the environment every day. It has been
reported that after the consumption of 80 million tons of coal, 1.35 million tons of
SOj is released into the atmosphere (Kumar and Prakash, 1977). As quoted by
Sharma (1986) 13 million tons offly ash, 4,80,000 tons of SO^, 2,80,000 tons ofNO^,
16, 000 tons of CO and 5,000 tons of hydrocarbons are released into the atmosphere
each year by our thermal power plants as long back as 1980. Further it has been
estimated that, every three tons of carbon burnt consumes eight tons of oxygen. That
is, we are borrowing from the present oxygen reserve of the atmosphere. With an
estimated known workable fossil fuel reserve of 8.6 trillion tons, man can deplete
about 17 X 10" tons of atmospheric oxygen (Sahu, 1994).
Toxic substances contained in flue gases discharged from stacks of thermal
power plants are potentially harmful to vegetation. Significant injury to living
vegetation can occur when atmospheric conditions are not conducive to rapid
dispersion of the pollutants. This injury can reduce the photosynthetic capacity of
the vegetation resulting in lower yields of green plant products and, in severe cases.
death of sensitive species. The main culprit of this in coal smoke is SO . The
devastating effect of SO^ on vegetation is well established and proved beyond doubt
(Katze/a/., 1939; Whitby, 1939; Thomas, 1951;Pelz, 1956; Thomas and Hendricks,
1956; Thomas, 1961; Linzon, 1965; Prokopiev, 1965; Rao and Le Blanc, 1966;
Dochinger, 1968;Stern, 1968; Jacobson and Hill, 1970; Jones e/a/., 1974;Muddand
Kozlowski, 1975; Mansfield, 1976; Kumar, 1977; Hallgren, 1978;MalhotraandBlanel,
1980;Dubeye^flf/., 1982; Iriving and Miller, 1984;Treshow, 1984; Malhotra and Khan,
1984;Heggestade/a/., 1986;Farooqe/fl/., 1988; Amundson^/a/., 1990; Murray and
Wilson, 1990; Sharma and Prakash, 1991;PolleeM/., 1992; Efee/a/ . , 1993;Qifuer
al, 1993; Huang a/., 1993).
A team of environmental botanists working at the department of botany,
Aligarh Muslim University, Aligarh, has studied the adverse effects of vegetation
around a thermal power plant, and found to affect some timber trees (Khan, 1982;
Ghouseefa/., 1984a,b; 1986a; Ahmad and Kalimullah, 1986,1988;Kalimullahcra/.,
1987; Guptas/a/. , 1988), Vegetable Crops (Amani and Ghouse, 1978; Gupta, 1981;
Khan and Khan, 1991) and the various weeds of the locality (Amani et al, 1979a,b;
Amani, 1982; Ghouse and Khan, 1983, 1984,1986;Khan, 1985; Ghouse and Saquib,
1985; Iqbal et al, 1986a,b; 1987a, b; Mahmooduzzafar et al, 1986, 1987; Saquib,
1989;Usmani, 1990;Malibarie/a/., 1991; Saheede/a/., 1993 a; Lone era/., 1994b).
Further, morphological variations, variation in the leaf epidermal architecture, sulphur
catching capacity as well as changes in the amount ofphotosynthetic pigments of different
plant species have also been studied under the influence of coal smoke pollution (Ghouse
etal, 1980; Khan and Khair, 1984, 1985a,b; Khan e/fl/., 1984; Ghouse a/., 1985
Ghouse and Saquib, 1986; Ghouse etal, 1986b; Gupta and Ghouse, 1986, 1987b
Saquib etal, 1986; Ahmad a/., 1987; Khan and Ghouse, 1988; Khan and Usmani
1988; Ghouse et al, 1989; Khan et al, 1991; Saquib and Ahmad, 1991; Lone and
Ghouse, 1992; Ghouse era/., 1993;Lonee/a/., 1993; Lone and Ghouse, 1993;Saheed
et al, 1993b; Lone et al., 1994a,b) and the biochemical impact on some plantation crops
(Lone, 1993). Measuring a physiological response to SO^ exposure under field conditions
is an important step in understanding the responses of trees to fumigation episodes and
in further predicting the effect on trees under field conditions. Regardless of several
reports on Indian crops and other plant species, the literature concerning the biochemical
responses of perennial broad-leaved forms under the stress of coal-smoke pollution in
field conditions is very meagre, and is yet to be carried out in detail. Therefore, the
present study has been directed in this line in order to unreel the biochemical behaviour
of some perennial species under the stress of coal-smoke pollution. The study will
elucidate the seasonal variations in the mineral balance, photosynthetic pigments, ascorbic
acid and proline, as well as some metabolic products like carbohydrate and proteins etc.
in the leaves of various selected species. Furthermore, the newly formed bark and wood
samples of the selected species have also been designed for analysis subjecting several
biochemical parameters. The experimental site for the present study has been selected
around a coal-fired power plant situated at Kasimpur, 16 kms. North-East of Aligarh City
in Uttar Pradesh.
The massive plantation drive launched in this country requires a proper
selection of species for plantation in areas which are under air pollution threats. For
this purpose, an effective screening of trees is essential, rather than seasonal crops
due to the long standing experience of trees to ambient pollution load. Trees are
already known to act as biological scavengers of toxic elements emanating out of coal
burning. They are capable of fixing atmospheric metallic Jlnd non-metallic oxides
present in ambient air. It is, therefore, believed to be essential to investigate the
common tree sj^cies^for their relative capability to fix the toxic elements by wide
screening techniques. The present investigation is undertaken with the hope that it
will yield useful data for an overall assessment of species for its performance and
suitability to plant in large number to provide green cover and biological purifiers to
the site.
Q'EOgfK^^ICSlL S'ET-'ltP
GEOGRAPHICAL SET-UP OF THE STUDY AREA
The Aligarh district lies in the North-West of Uttar Pradesh, a Northern state
of India, in the fertile agricultural area of Ganga Jamuna Doab between 27° 29 N and
28° 11 N latitude and 77° 29 E and 78° 38 E longitude (Fig. 1). Kasimpur, the site of
Thermal Power Plant Complex, lies in the Morthal Pargana of the Koil Tehsil in the
Aligarh district. On the northern border of the town, flows the Upper Gangetic Canal
supplying water to the power plant. This place is about 16 km. (road distance) in the
north-east of Aligarh city (Fig. 2) situated between 27° 59 N and 28° 3 N latitude and
78° 8 E and 78° 93 E longitude, about 187 meters above the sea level.
METEOROLOGY
The study area experiences a dry and tropical monsoon type of climate with
seasonal rhythm marked by the north-east to south-west monsoon. The year com-
prises three principal seasons viz. cold weather season (winter), hot weather season
(summer) and rainy season (monsoon).
The Cold Weather SeasonAVinter (November to March)
The beginning of winter season is marked by a considerable fall in tempera-
ture. In this season, a relatively low pressure exists over the Indian seas, thus causing
the winds to blow from plains towards the seas. The mean maximum temperature is
27.11°C inNovember and 30.14°C inMarch, while the average minimum temperature
for these months is 12.98°C and 14.63°C respectively (Fig.3). It is very cold in the
month of January (7.87°C - 21.32°C), the temperature begins to rise (14.63°C-
30.14°C) in March. In winter months the nights are very cold and the days are
comparatively warmer with foggy mornings.
Wind direction during the winter season is predominantly from east to west.
KOIL TAHSIL ( ALIGARH )
N
{»axj RAILWAY TRACK
= 1 UPPER 6AN6A CANAL
Z l j ROAOS
^ H i j EXPERIMENTAL SITES
3 KM U-1
FIG. 2
Fig. 3 The graph shows monthly temperature in centigrade (Average of years 1991-1993)
Tomperaturo ( C)
Nov Dec Jan Feb Mar Apr May Jun Jul Aug Sep Oct Winter Summer- - Mansoon
— ^ Minimum - + - Average Maximum
west to east or south east to noth east. The winds during this season are light and blow
at an average speed of 3.06 km/hour (Fig.6A). The average relative humidity during
this season is 49%, 57.5%, 61%, 52% and 38.5% for months November to March
respectively (Fig.5). The rainfall in this season is irregular and sporadic (Fig.4).
The Hot Weather Season/Summer (April to June)
This season extends from March to June. It begins with an appreciable rise in
temperature and a decrease in pressure. Due to wide range of temperature during the
summer months, days are warm and nights are pleasant. The minimum and maximum
temperatures in April are 19.01°C and 37.01®C respectively. The temperature
continues to rise during May (23.29''C-39.50»C) and June (26.60''C-40.86»C) (Fig.3).
Days are hot and dry, the average relative humidity declining to 30%, 35% and 44.5%
in the months of April, May and June, respectively (Fig.5). The hot day winds
blowing with high velocity form a regular phenomenon. The velocity of wind begins
to increase steadily from April with an average wind speed of 4.31km/hour (Fig.6-
B). The wind speed rapidly increases during 8 am to 1 pm causing the wind to blow
almost with the force of gale during the next 2-3 hours. It then falls suddenly by 6.
pm and a calm usually prevails during the night.
Dust and thunder storms are frequent, at times accompanied by rains. The
rains are rare, sporadic, short lived and highly variable in amounts. The average
monthly rainfall is about 0.75 mm for April, 14.4 mm for May and 23.60 mm for June
respectively (Fig.4).
The Rainy Season/Monsoon (July to October)
The atmospheric temperature falls with the arrival of the humid oceanic
currents and the air becomes cool and pleasing by the end of June. The average
minimum and maximum temperatures fall to 24.84°C - 34.48"C in July, 24.79"C -
Fig. 4 The graph shows monthly rainfall in mm (Average of years 1991-1993)
300 Rainfall (mm)
Nov Dec Jan Fab Mar Apr May Jun Jul Aug Sep Oct Winter Summer - - Monsoon--Maximum Average Minimum
Fig.5 The graph shows monthly relative humidity In percentage (Average of years 1991-1993)
100 Relative humidity («)
Nov Dec J&n Feb Mar Apr May Jun Jul Aug Sep Oct Winter Summer - - Monsoon--Maximum Average Minimum
( s c a l e 1 C i n - 5 V . )
N
3.if7
3 . 0
3 .57
3 . 0 7
0 1.88 2 .85
FIG. 6 A : VELOCITY « PERCENTAGE OF WIND DIRECTION DURING WINTER SEASON
5'32
h.22 I3.6f»
5.23 3 . 0 8
F I G . 6 B : VELOCITY & PERCENTAGE OF WIND DIRECTION DURING SUMMER SEASON
3.60
3.6«f 1 3 -66
3 . 6 0 f colm ^ 3.U6
i».58 1 3 . 0 2
2 .71
F IG . 6 C : VELOCITY ^ PERCENTAGE OF WIND DIRECTION DURING MONSOON SEASON
33 .38°C in August, 22.54''C- SS-eCCin September and I8.46''Cto 33.47''Cin October
(Fig. 3). The average relative humidity is 72%, 75%, 70% and 52% for t h ^ e months / \
respectively (Fig. 5). The sky is generally overcast. Rain set isusually by the end of June
or early July and continues untill the end of September or early October. The average
maximum rainfall (about 191 mm and 177.5 mm) was observed in the months of July and
August respectively (Fig.4). During this season winds blow pre-dominantly from east-
west, to west-east, and to south-west to north-east, to south-west and to south-e^st to
north-west (Fig.6C) with an average speed of 3.54 km/hour.
SOURCE OF POLLUTION
The thermal power plant complex of Kasimpur was found to be the source of
pollution. The complex, one of the three major thermal power plants of Uttar Pradesh
is located along the banks of the Upper Gangetic Canal running in the north-west to
south-east direction and consists of three power stations (A,B and C) having a
capacity of 90 MW, 210 MW and 230 MW electricity generation respectively (Plate-
I). The whole complex runs on the low grade coal transported from various collieries
of North India. The chief chemical constituents of coal are 2.92% moisture, 22.20%
ash, 31.86% volatile matter, 0.49% sulphur, 5.61% hydrogen, 5.24% nitrogen,
20.23% oxygen and 42.45% fixed carbon on an average (Table-1).
The data (Table-2) on coal combustion at the above source shows that annual
coal consumption comes to about 1,28,544 metric tons for plant 'A ' , 5,35,971 metric
tons for plant 'B' and 8,66,200 metric tons for plant ' C . The total annual coal
consumption in all the power plants comes to be about 1,5 3 0,715 metric tons.The daily
average coal consumption is about 4194 metric tons. The average monthly and daily coal
consumption during the different seasons (winter, summer and monsoon) is given in table
(2). On the other hand, the unburnt coal possess considerable amounts of sulphur,
nitrogen and carbon and when subjected to 1200''C - MOO^C high temperature for its
1
PLATE-I
ABOVE} Pliotogir«|)h ihovtt tht atalti-it&«k ^mtt rumt
B lLOWt jPhofogmiKfiilMMri tlic ttiick liNkt
{itti stiKk) iitd ^C* (rtgtii mtkS
Table 2. Data on coal consumption (in metric tonnes) in the Thermal Power Plant Complex ofKasimpur (An average of data forthe years, 1991-1993)
Months Power Stations Total Monthly Consumption
A B C
*
Nov 08530 42650 72059 123239 Dec 09240 53153 83674 146067 Jan 10992 48070 93579 152641 Feb 12830 38128 78697 129655 March 18941 56674 85688 161303 Apnl 14338 52810 72844 139992 May 12665 53252 67929 133846 June 08627 38382 58957 105966 July 07920 26623 56011 090554 August 07322 38403 63050 108775 Sept 08899 42852 66602 118353 Oct 08240 44974 67110 120324
Total m Wmter (Nov - March) 60533 00 238675 00 413697 00 712905 00
Monthly average 12106.60 47735 00 82739 40 142581 00
Wmter daily average 400 88 1580 63 2739 15 1953 16
Total m Summer (Apnl - June) 35630 00 144444 00 199730 00 379804 00
Summer Monthly average 11876 67 48148 00 66576 67 126601 33
Summer daily average 391 54 1587 30 2194 84 4173 67
Total in Monsoon (July - Oct) 32381 00 152852.00 252773 00 438006 00
Monsoon Monthlv average 08095 25 38213 00 63193 25 109501 50
Monsoon dail> average 263 26 1242 69 2055 00 3561 00
Annual consumption 128544 00 535971 00 866200 00 1530725 00
A\'erage monthl) consumption 10712 33 44664 25 72183 33 127559 58
A\'erage daiK consumption 352 17 1468 50 2373 15 4193 74
Source Courtse> of AEE, Thermal power plant complex of Kasimpur
Table 3. Amount of gases released from the Thermal Power Plant Complex in different Months (Average of three years, 1991-1993).
Months Amount of SO^ Amount of NO^ Amount of CO^ Months
Kg/hr ppm/hr Kg/hr ppm/hr Kg/hr ppm/hr
Nov 16437 0016 294416 0 294 7784993 07 785 Dec 18848 0019 337696 0 338 11294414 11 294 Jan 19695 0 020 352894 0 353 14647129 14 647 Feb 18523 0019 331869 0 332 14550399 14 550 March 20814 0 021 372920 0 373 17809853 17810 Apnl 18675 0019 334590 0 335 1819826 01 820 May 17278 0017 309442 0 309 2801652 02 802 June 14129 0014 253152 0 253 2292021 02 300 July 11684 0012 209354 0 209 1895479 01 900 Aug 14503 0015 259862 0 260 2352779 02 353 Sep 15271 0015 273623 0 274 2477369 02 477 Oct 15533 0016 278180 0 278 2519855 02 520
Average in Winter (Nov - March)
18863 0 0190 337959 0 338 13217358 13 217
Average in Summer (Apnl - June)
16694 00170 299061 0 299 2304500 02 307
Average in Monsoon (July - Oct)
14248 0 0145 255255 0 255 2311370 02 312
Average 16782 00169 300666 0 300 5944409 05 945
Source CourtscN of AEE, Thermal power plant of Kasimpur
combustion, obviously produces noxious gases like SO , NO^ and CO^. These gases
along with other particulate pollutants spread into the atmosphere through various stacks.
The computed amounts of SO^, NO^ and CO^ being released into the atmosphere in kg.
per hour and ppm per hour are given in Table-3.
SOIL AND EDAPHIC CONDITIONS
The soil at the test site for the present study has a structure categorised by the
Agricultural Directorate (Soil Survey and Research, Aligarh) as type III (Fig.7).
This consists of the loam and clayey loam type of soil with a very high pH value and
very poor drainage system. During monsoon season it suffers from water Jogging.
The clay content is maximum at the top and decreases with depth. It is ash grey in
colour tending to become black when moist. The poor drainage results in the
deposition of soluble sodium salts on the surface in the form of 'Reh' . During drought
periods, the land becomes white and salt infested. The soil is mostly of alkaline in
nature. The data obtained on the mechanical analysis as well as the physico-chemical
properties of the soil collected from the test site as well as from the reference site
during the study period is given in Table-4. It is clear from the data that the sand
fraction at both the sites is below 50%. The coarse silt happens to be higher in
percentage than the fine silt at both the places. Further, the clay percentage in soil
is slightly higher at Aligarh site than at Kasimpur.
The soil pH at both the sites is alkaline, however, the alkalinity is higher at
Aligarh site than at kasimpur. Further, there was a linear increase in soil pH at both
the sites from summer to winter seasons. Total nitrogen as well as available
phosphorus contents were recorded lesser at the polluted site compared to non-
polluted site. On the other hand exchangeable potassium was same in polluted as well
as control sites in summer season. However, slightly lesser contents were observed in
monsoon and winter seasons at the latter site. Sulphur was found in higher amounts at the
polluted site, the maximum contents being observed in monsoon season. Similarly,
organic carbon was also recorded higher at the pollutd site than at the control.
7
KOIL TAHSIL SOILS
k - i TYPE I I I
m i l TYPE IV
m i USAR
t-.-i KHADAR
SITES 0 3 ' • • '
KM
FIG. 7
Tflbic 4. Mechanical and physico-chemical analysis* of soil samples collected from the polluted and control sites during the study period.
Mechanical Analysis Site P Site C
1 Sand(%) Coarse Fine
2 Silt (%) Coarse
Fine
3 a a y ( % )
4 Texture Class
Physico-Chemical Analysis
1 Porosity
2 Water Holding Capacity
3 E C (m mhos/gm)
4 C E C (m eq /g soil)
1 07 48 50
16 93 10 70
22 80
Sandy Clay Loam
46 65
39 10
4 30x10 3
14 30
1 87 42 21
14 90 11 82
29 90
Sandy Clay Loam
43 24
35 49
3 33x103
04 10
SUMMER MONSOON WINTER
Si te -^ P C P C P C
5 pH 7 60 8 00 7 85 8 17 7 87 8 20
6 Total N(%) 0 070 0 076 0 060 0 063 0 068 0 072
7 Available P(%) 0 080 0 085 0 110 0 120 0 130 0 140
8 Exchangeable K(%) 0 077 0 077 0 077 0 075 0 077 0 076
9 Sulphur (ng/gm) 500 285 525 290 520 290
10 Organic C (%) 4 10 1 67 5 10 1 90 5 00 1 90
Site P= Polluted Site (Kasimpur) Site C= Control Site (Aligarh) ' Soil analysis earned out at the Agncultural Directorate (Soil Surve) and Research), Aligarh
fAOllt^HilSlLS
fKCET^CCXDS
MATERIALS AND METHODS
SELECTION OF THE EXPERIMENTAL SITES
Under these ecological and meteorological conditions (As described in earlier
chapter) the test site for the present study was maintained within I km. from the pollution
source in the down wind direction. The Botanical Garden of the Botany Department of
Aligarh Muslim University, Aligarh served as a control site situated at a distance of 16 km.
in the cross wind direction from the test site and free of any coal-smoke pollution (See
Fig. 2).
SELECTION OF THE SPECIES
After a general survey, the following species growing in open land were
selected at the rate 5 per site.
1. Botanical Name = AZADIRACHTAINDICA A. IIJSS
English Name = NEEM TREE, MARGOSA TREE
Family = MELIACEAE
Age = 25 YEARS
This is a medium-sized tree with a clear bole of 10-25', and a girth of 6-8', with
a large crown. Bark, grey or dark grey or nearly black; leaves, imparipinnate,
alternate, bluntly serrate. Flowers white or pale yellow, small, honey scented. The
tree is evergreen, first flush of growth starts in the begnning of March and the first
collection was made in the first week of April. It is commonly found throughout the
greater part of India, but wild on the east cost and the Deccan; also in Siwaliks; very
often cultivated. Grows on almost all kinds of soils, but does well on black cotton
soil. Almost every part of the tree is bitter and has found application in indigenous
8
medicine. Timber pest proof used for furniture, agricultural implements, board,
pencils, small articles etc. The tree is well suited for afforestration in dry situations
and is a common avenue tree.
2. Botanical Name = TAMARINDUS INDICAUnn.
English Name = TAMARIND-TREE
Family = CAESALPINIACEAE
Age = 25 YEARS
A moderate-sized to large, evergreen shade tree, with handsome, dense
crown, stem short, thick, seldom straight. Bark, brownish or dark grey, longitudi-
nally and horizontally fissured. Leaves, paripinnate; leaflets generally 10-20 pairs,
subsessile; flowers, small, yellowish with pink stripes. Fruits, greyish-brown green,
more or less constricted between seeds, slightly curved. Growth activity starts by the
first week of March and the first collection was made in the first week of April. The
tree is indigenous to tropical Africa, cultivated or found naturalized almost through-
out the plains and Sub-Himalayan tracts of India, particularly in the south. Largely
planted on road sides. Grows from the coast to the hills, young plants grow best on
porous soil. It is a good source of tartaric acid, malic acid, and other important
vitamins and minerals. Timber used for agricultural implements.
3. Botanical Name = CASSIA FISTULA Unn.
English Name = INDIAN LABURNUM, GOLDEN SHOWERS
Family = CAESALPINIACEAE
Age = 30 YEARS
A deciduous, medium-sized tree with a bole of 12-15' high and 3-4' in girth.
Indigenous to India, and naturalized in tropical Africa, South America and West
Indies. Leaves, 20-40 cm, glandless; leaflets 4-8 pairs. The first flush of growth
starts from the beginning of April and the first collection was made in the second
week of May. Bark, grey, smooth, exfoliating in small woody scales. The fruits are
pendulous, cylindrical 25-50 cm long. Leaves are antiperiodic, laxative and used in
jaundice, piles, rheumaticism, ulcers and skin-eruptions. Fruit pulp rich in protein
and carbohydrate, laxative and antibiluous; given in diabetes and blood-poisoning.
Timber is a suitable substitute for teak and sal. The tree is excellent for agro-and
social forestry. Reported to fix nitrogen and used as green manure.
4. Botanical Name = FICUS BENGALENSISUnn.
English Name = BANYAN TREE
Family = MORACEAE
Age = 25 YEARS
A very large tree with spreading branches, evergreen, attaining at times the
height of 100'; aerial roots many. It attains large dimensions, the leafy crovirn
sometimes attaining a circumference of 1,000 - 2,000'. Leaves 4-8 in long coria-
ceous, ovale to elliptic with round or subcordate base; fruits sessile in pairs. Various
parts of the plant are considered as medicinal. The growth activity starts in the
middle of March a'-.d the first collection was made in April. The bark contains 11%
tannin. The wood is grey or greyish white and moderately hard. It is durable under
water, the wood of aerial roots is stronger and more elastic. The tree occurs
throughout the forest tracts of India. It is grown in gardens and road-sides for shade.
The banyan tree is one of the recorded host of Indian lac insects.
10
5. Botanical Name = FJCVS RELIGIOSA Unn.
English Name = PEEPAL TREE
Family = MORACEAE
Age = 25 YEARS
A large evergreen fast growing tree, epiphytic when young, with spreading
branches and round or broadly ovate, caudate, more or less pendulous leaves; Bark,
rough, white to grey, fruits sessile in auxiliary pairs, depressed globose. The fruits
and tender buds are occasionally eaten in time of scarity. The leaves and twigs are
lopped for cattle fodder. The first flush of growth starts at the end of March,
accordingly the first collection was made in the last week of April. The tree yields
a latex, and the various parts of the plant are considered as medicinal. The bark
contains 45% tannin. The tree is found wild or cultivated rarely throughout India and
is held sacred by Hindus and Buddhists. It is planted as an avenue or road side tree,
and is also one of the recorded host of the Indian lac insect in Madhya Pradesh,
Bengal and Assam.
SAMPLES AND SAMPLING TECHNIQUES
For each species 5 individuals of almost similar age group were selected at each
site and from each tree newly emerged 15 days old leaves were sampled in the months
of March to May (Sumiror season), depending on the time of initiation of extension
growth. The leaves with an apparently healthy look were collected very carefully from all
the sides of the tree crown as well as from the top and bottom of the canopy. About 100
samples were collected from each tree, mixed together and thus it served as one replicate.
Similarly, the same number of leaves were collected for all individual representatives and
the samples collected from 5 trees served as 5 replicates for each species at each site. The
sampled leaves were packed in polythene bags and brought to the laboratory in ice
11
boxes. Before being, processed for any chemical analysis the leaves were gently cleaned
with moist cotton to remove any particulate matter deposited on their surfaces. Except for
chlorophyll, protein, proline and ascorbic acid estimations where the fresh material was
used, the samples were oven dried for other biochemical studies.
The subsequent samplings for similar studies were made in monsoon, and late
winter seasons. It was insured that the foliage of last two collections represent the
product of first flush and the foliage developed in the subsequent flushes, if any, was
carefully discarded. Bark and wood samples were also collected from the experimen-
tal trees in summer, monsoon and winter seasons, along with the foliage. From each
selected individual, two blocks of about 2 cm thick were chiseled out of the main
trunk at chest height (1.5 meters from the ground) and each block containing
sufficient amount of bark and wood portions. The sampled blocks were processed
with stainless steel penknife to separate carefully the portions of wood and new bark.
The older and apparently dead parts of the bark were discarded. Thus the sundered
wood and bark portions were chopped off in fine slices and sufficiently dried in an
oven at 80°C. The bark and wood samples of both the blocks of each replicating
individual were mixed and ground in a laboratory grinder to a fine powder and used
for desired analysis. The grinder was thoroughly washed and cleaned after every use
to avoid contamination.
PARAMETERS
The samples of leaves, bark and wood of the selected species were analysed on
a comparative basis for sulphur, photosynthetic pigments, carbohydrate, protein, nitro-
gen, phosphorous, potassium, sodium, calcium as well as ascorbic acid and proline
contents. The observations recorded on these parameters have been discussed seperately
under different heads.
12
STATISTICAL ANALYSIS
The data were analysed by Split-plot design by the method mentioned by Sir
S.A. Fisher (Dospekhov, 1984). The correlation coefficient (r) has been calculated
between sulphur and other parameters.
METHODOLOGY
The following procedures has been followed for the estimation of
various biochemical parameters in the present study.
SULPHUR
For the estimation of Sulphate-Sulphur, the method givenby Patterson (1958)
was adopted. The oven dried samples of leaves, bark and wood were ground and
passed through 72 mesh screen. 300 mg screened powder and 0.1 ml selenium
dioxide (SeO^) solution was digested using 10 ml conc. HNOj and 1 ml of conc. HCl.
After filtering the digested material, 10 ml of 3% glycerol was added and volume
made upto 100 ml with distilled water. To this solution 5 ml of 2% barium chloride
(BaClj) solution was added to precipitate sulphur as barium sulphate (BaSO^). The
optical density was noted at 420 nm on a spectrophotometer. The amount of sulphur
was determi^.ed by freshly prepared standard curve with potassium sulphate solution.
PHOTOSYNTHETIC PIGMENTS
The fresh leaf samples after being brought to the laboratory were removed
from the ice bags and their surface gently cleaned with the moist cotton to remove
any particulate matter deposited over them. The chlorophyll content was estimated
by following the method of Arnon (1949). One gram fresh sample was crushed gently
13
with 80% acetone in a mortar and pestle. To this was added a little pinch of calcium
carbonate (CaCOj). The samples after being ground to a fine pulp were centrifuged
(5000 rpm for 5 minutes) and the supernatant transferred to a 100 ml volumetric
flask. The residue was repeatedly ground and centrifuged till it turned colourless,
thus ensuring the complete extraction of chlorophyll from the tissue. The volume of
the extract was made 100 ml by adding 80% acetone. The absorption of the solution
was read at 663, 645, 510 and 480 nm on spectrophotometer. The chlorophyll (a and
b) and carotenoid contents were analysed by applying the formulae given by
Maclachlan and Zalik (1963) and Duxbury and Yentsch (1956), respectively. The
total chlorophyll was estimated by applying the formula given by Arnon (1949). 12.3 D663 - 0.86 D645
mg chlorophyll a/g tissue = x V d X 1000 x W
19.3 D645 - 3.60 D663 mg chlorophyll b/g tissue = x V
d X 1000 X W
7.6D480- 1.49 D510 mg carotenoids /g tissue = x V
d x 1000 x W
20.2 D645 - 8.02 D663 mg total chlorophyll/g tissue = x V
d X 1000 X W
Where D663, D645, D510 and D480 represent the values of optical densities
at the respective absorption spectra.
V = final volume of chlorophyll extract in 80% acetone.
W = fresh weight of tissue extracted,
d = length of light path.
14
CARBOHYDRATE
The total carbohydrate contents of leaves, bark and wood samples was
estimated by following the method of Dubois etal. (1956) (Sadasivum and Manikam,
1992). 100 mg of the dried sample was taken in a boiling tube and hydrolysed by keeping
it in a boiling water bath for three hours with 5 ml of 2.5N-HC1 and cooled to room
temperature. Solid sodium carbonate was added to this solution to neutralize it until the
effervescene ceased. The volume of the solution was made up to 100 ml and then
centrifuged. Then 0.1 and 0.2 ml ofthe sample solution was pipetted out in graduated
tubes and final volume made upto 1 ml with double distilled water. 1 ml of 5% phenol
solution was added to each tube followed by 5 ml of 96% H^SO^ and shaken well. The
test tubes with solutions after 10 minutes were then placed in water bath at 25-30°C for
20 minutes and the colour read at 490 nm on "Spectronic-20" colorimeter. A blank was
run with each sample. The carbohydrate content was calculated by comparing the optical
density of the sample with a calibration curve plotted by taking known dilutions of
standard solutions of chemically pure glucose.
PROTEIN
The protein contents in the samples was estimated by following the method of
Lowry^/a/. (195 l)(Sadasivam and Manikam, 1992). One gram offresh leaf sample (for
bark and wood dry samples were used) was homogenised with 5-10 ml of phosphate
buffer (pH 7-9) in a glass mortar. The solution was centrifuged and the supernatant used
for protein estimation. 0.1 ml and 0.2 ml ofthe sample extract was pipetted out in two
graduated tubes and volume made upto 1 ml in each tube. Another tube with 1 ml of water
served as blank. To each of these test tubes was added 5 ml of Reagent C (Appendix-
3.3). The solution was mixed well and allowed to stand for lOminutes. ThenO.5 ml of
Reagent D (Appendix-3.4) was added to this solution in each tube and incubated at room
temperature in dark for 30 minutes. The blue colour developed was read at 660 nm on
15
a "Spectronic-20" colorimeter. The protein content was estimated comparing the
optical density of each sample with a calibration curve plotted by taking known dilutions
of a standard solution of bovine serum albumin (Fraction V).
NITROGEN
The oven dried samples of leaves, bark and wood were powered and passed
through 72 mesh screen. The samples were then digested by following the method
of Lindner (1944).
Digestion of Samples
100 mg dry powder of the sample was taken in a 50 ml kjeldahl flask. Two
ml of pure H^SO^ was added and the mixture was heated for about two hours to
dissolve the powder. This acid turned the contents black. After cooling the flask for
about 15 minutes, 0.5 ml of chemically pure 30% hydrogen peroxide was added drop
wise. The solution was again heated about 30 minutes, until it turned light yellow
in colour and then cooled with 3-4 drops of H^Oj, it was reheated for about 15
minutes to get a clean extract. Excess of hydrogen peroxide was avoided which
would otherwise oxidise the ammonia in the absence of organic matter. The peroxide
digested material was transferred to 100 ml volumetric flask with three or four
washings with double distilled water (DDW) and the volume made upto mark. This
served as the stock solution foi the estimation of N, P and K.
Estimation of Nitrogen
A 10 ml aliquot of the peroxide digested material was transferred to a 50 ml
volumetric flask. To this, 2 ml of 2.5N sodium hydroxide was added to neutralize
the excess of acid partially. To prevent the turbidity, one ml of 10% sodium silicate
was added to the flask and the volume made up. In a 10 ml graduated test tube, 5
16
ml of aliquot of this solution was taken and added with 5 ml of Nessler's reagent
(Appendix-1.1) and followed by thorough shaking. The final volume was made up
with DDW and kept for about 5 minutes for the maximum colour development. This
solution was taken in a colorimetric tube and its optical density measured at 525 nm
on a "Spectronic-20" colorimeter. A blank was run simultaneously during determi-
nation. A standard curve of known dilution of ammonium sulphate solution was made
and the reading of each sample compared with the calibration curve. Nitrogen in each
sample was calculated in terms of percentage on dry weight basis.
PHOSPHORUS
The samples were digested by following the method of Lindner (1944) (as
described in case of Nitrogen) and phosphorus was estimated by the method given
by Fiske and Subbarow (1925). In a 10 ml graduated tube, 5 ml of aliquot of peroxide
digested material was taken and 1 ml Molybdate reagent (Appendix-2.1) was added
carefully, followed by 0.4 ml of Amino-naphthol-sulphonic acid (Appendix-2.2).
The colour of the solution turned blue and its volume made upto 10 ml with double
distilled water. The solution was shaken well and allowed to stand for 5 minutes for
maximum colour development. The solution was transferred to a colorimetric tube
and per cent transmittance was read at 620 nm, on a "Spectronic-20" colorimeter.
The standard curve was plotted by using known dilutions of monobasic potassium
phosphate solution.
POTASSIUM
The samples of leaves, bark and wood were digested by the method of Lindner
1944 (as described in case of nitrogen). The potassium contents in the samples were
estimated flame photometrically. One ml of aliquot (peroxide digested material) was
suitably diluted with double distilled water in a graduated tube. A blank containing only
17
distilled water was run simultaneously. The readings were compared with a calibration
curve plotted using the known dilutions of a standard potassium sulphate solution. The
potassium was expressed on per cent dry weight basis.
CALCIUM
The oven dried samples of leaves, bark and wood have powdered and passed
through 72 mesh screen. The samples were digested following the procedure given
by Singh (1988).
Digestion of Plant Materials
50 mg of the oven dried plant material was taken in a 50 ml volumetric flask.
Two ml of concentrated Nitric Acid (HNOj) was poured into it and heated on a hot
plate till brown effervescence was produced. At this stage, sufficient quantity of
TAM solution (Appendix-4.1) was added to make the solution clear and then heated
to dryness. After adding sufficient quantity of double distilled water (DDW), the
solution was transferred to another 50 ml volumetric flask with three washings. The
final volume was made upto the mark with double distilled water, the calcium content
of the samples was calculated on per cent basis by reading in a flame photometer with
the help of calcium filter. The readings were compared with a standard curve plotted,
using known dilutions of calcium bicarbonate.
SODIUM
The samples were digested by following the procedure given by Singh (1988)
(as described in case of Calcium). The digested samples of leaf, bark and wood was
taken to a flame photometer and the sodium content was read with the help of sodium
filter. The per cent quantity of the element was calculated by comparing the readings
on a standard curve plotted by using known dilutions of sodium chloride.
18
ASCORBIC ACID
The ascorbic acid content of leaves was determined colorimetrically by using
the 2,6-dichlorophenol Indophenol (DC PIP) method (Keller and Schwager, 1977).
0.5 gm leaf was homogenised in a solution prepared by dissolving 0.5 gm oxalic acid
and 0.015 gm EDTA. The homogenate was then centrifuged at 3000 rpm for 15 min.
To one ml of supernatant 0.5 ml of 2, 6 DC PIP was added with a constant shaking
at Ajj^ of pink colour was recorded (Es). To this pink colour 1 per cent aqueous
solution of ascorbic acid was added in order to remove pink colour and again A ^ was
recorded (ET). The blank was prepared with distilled water and 2, 6 DC PIP and A ^ ,
was recorded (Eo) with "Spectronic-20" spectrophotometer. Amounts were deter-
mined with the help of a calibration curve prepared by using one per cent ascorbic
acid solution.
Eo-Es-ET Ascorbic acid mg/gm fresh leaf = x F
W
Where, W = weight of leaf
F = Factor (Calculated from Calibration Curve)
The amount of ascorbic acid expressed in the table is mg/100 gm fresh sample.
PROLINE
The free proline content in leaf fresh weight was identified by means of a
colorimetric method using ninhydrin, according to Bates et a/.,(1973). 0.5 mg leaf
samples were extracted by homogenizing in 10 ml of 3% aqueous sulphosalicyclic
acid and filtered the homogenate through Whatman No. 2 filter paper. From this, 2
ml of filtrate was taken in an another test tube added 2 ml glacial acetic acid and 2 ml acid-
ninhydrin to it. Then, test tubes were transferred into a boiling water bath for 1 hr. The
19
reaction was terminated after 1 hr. by placing the tubes in an ice bath, added 4 ml toluence
to the reaction mixture and stirred well for about 20-30 sec. The toluence layer was
separated and warmed it to room temperature. The colour intensity (red colour) was read
at 520 nm. In a similar way a series of standard was prepared with pure proline (Zigma
proline) and plotted a standard curve. The amount of proline in the test sample was
calculated from the standard curve and expressed in fiMg' fresh weight of leaves as a
mean value from five replicate calculations.
20
(yBS'E^iVTlOh^
OBSERVATIONS
SULPHUR IN LEAVES
The data, on the effect of coal-smoke pollution in relation to sulphate contents
of the leaves are summarized in Table-5. The data show that all the investigated
species have recorded an increased amount of sulphate-sulphur in their foliage under
the pollution stress.
The Fig. 8 indicates the seasonal trend in the accumulation of sulphate-sulphur
in the foliage of the investigated species. It becomes clear from the figure that, all
the investigated species have accumulated the peak amount of sulphur in the
monsoon season, irrespective of the pollution level with the exception of Cassia
fistula in which the peak occurs in winter.
The per cent variation given in Fig. 11 shows that the variation in sulphate-
sulphur content happens to be positive in the polluted population of all the
investigated species irrespective of seasonal conditions. The variation in the
inorganic sulphur content happens to be the highest in Azadirachta indica (Winter
collection) which is closely followed by Ficus religiosa (Monsoon collection),
Ficus bengahftsis and Cassia fistula (Summer collection), while in Tamarindus
indica, the variation ranged from 40.67% to 45.89%, the highest occurring in
monsoon.
The seasonal mean of sulphate-sulphur occurrence given in Fig. 11 reveals
that all the investigated species have accumulated increased percentage of sulphate-
sulphur in their foliage in the SO^ enriched environment and can be arranged in
decreasing order as under: F. religiosa (83.43%), F. bengalensis (82.26%), C.
//5/i//a (80.51%), 7W;ca (73.53%) and T. indica (43.06%).
SULPHUR IN BARK
The sulphate-sulphur contents of the newly formed bark tissues on analysis
> ^ it) OS
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A
B O es 2
b. B
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I M W
T.indica, 0 8 '
0 7 -
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F.religiosa.
i l M W
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F.bengalensis.
J 1 i i B Control • Polluted
S - Summer M - Monsoon
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W - Wuiter
Fig.8. Seasonal variation in the contents of sulphate-sulphur (per cent dry weight) in the leaves of various trees species.
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0 3 0
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T.indica.
j l M M W
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F.religiosa.
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B Control • Polluted
W
0 4 0
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0 3 0
0^ 0^, 0 1 5 ,
010.
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F.bengalensis.
l l l l i L
S - Summer M - Monsoon
M W
W - Winter
Fig.9. Seasonal variation in the contents of sulphate-sulphur ( per cent dry weight) in the bark samples of various trees species.
show that it accumulates significantly in the bark tissues of all the investigated
species under the stress of coal-smoke pollution, except T. indica in which the
accumulation has been found to be non-significant (Table-6).
The seasonal changes in the amount of sulphate-sulphur content in the bark
is given in Fig. 9. It indicates that the highest amount of sulphur occurs in monsoon
in the case of C. fistula and F. religiosa, in summer in case of A. indica, T. indica
and F. bengalensis. However, the interseasonal variation in C. fistula and Ficus
spp. has been found to be statistically non-significant. The per cent variation noted
over the control on sulphate-sulphur contents (Fig. 11) shows that the newly formed
bark tissues of all the investigated species express positive variation over the control
in their sulphate-sulphur contents in all seasons under pollution stress. The per cent
variationin sulphate-sulphur content is wider in F. bengalensis (59.01%) occurring
in monsoon collection and it is followed by C. fistula (41.81%) occurring in winter,
F. religiosa, A. indica and T. indica occurring in summer.
The seasonal mean of sulphate-sulphur content in bark of the investigated
species given in Fig. 11. It reveals that, the bark samples of all the investigated
species have accumulated significant amount of sulphate under pollution stress
except T. indica in which the accumulation has been found to be non-significant.
Among them, the highest percentage of accumulation has been observed in the bark
samples of Z'. bengalensis (54.83%), followed by C. fistula (38.84%), F. religiosa
(37.62%) and A. indica (28.87%).
SULPHUR IN WOOD
The results obtained in the present study on the sulphate-sulphur contents in
the sap wood samples of the investigated species show that in all the investigated
species, the level of sulphate in the sap wood samples increases under the influence
of coal-smoke pollution. But, it happens to be only marginal and non-significant in
the case of T. indica and F. religiosa (Table-7).
22
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II II II S M w
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J l U l S M W
B Control • Polluted
040-
0 3 5 .
0 3 0
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010.
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F.bengalensis.
a l i l • ! M
S - Summer M - Monsoon
W
W - Winter
Fig.lO. Seasonal variation in the contents of sulphate-sulphur ( per cent dry weight) In the wood samples of various trees species.
o o o C> o 7 q* 7 ° T o o
A O O
I
c I
i
I
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(s. < I-)
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The seasonal variation in the sulphate-sulphur contents is represented by Fig 10.
The figure indicates that, in A. indica, T. indica and F. religiosa, the amount of
accumulation reaches the peak in monsoon and slightly falls in winter, where as in C.
fistula and F. bengalensis the amount of sulphate-sulphur increases in sap wood
samples from summer to winter in a gradual manner. However, the interseasonal
variation occurred in all the investigated species has been found to be non-significant,
irrespective of seasonal as well as pollution level.
The per cent variation worked out over the control on the basis of their
sulphate-sulphur contents, given in Fig. 11 implies that, in all the species studied
there has been a positive variation in sulphate-sulphur contents in their sap wood
samples. The variation being the widest in summer in A. indica (17.89%), C. fistula
(55.69%) and F. religiosa (8.94%), while in F. bengalensis the maximum variation
occurs in monsoon (21.73%) and T. indica in winter (6.25%).
The seasonal mean of sulphate-sulphur contents included in Fig. 11 shows that
the significant amount of sulphate-sulphur accumulation has occurred in the sap
wood samples of mt/zca (14.56%), C. fistula .\1%) and F. bengalensis^
(17.39%), while in T. indica and F. religiosa it has been found to be only marginal
and non-significant.
PHOTOSYNTHETIC PIGMENT
The data (Table 8-12) cn the effect of coal-smoke pollution on the
photosynthetic pigments in the foliage of the investigated species reveal that, in all
of them the amount of photosynthetic pigments (Chl.a,b, carotenoids and total
chlorophyll) undergoes a significant loss under pollution stress, except in F.
bengalensis, in which the photosynthetic pigments exhibit a significant gain in the
SOj enriched atmosphere.
The seasonal alteration in the level of photosynthetic pigments given in Fig.
23
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Fig.l2. Seasonal changes of photosynthetic pigments (mg/g fresh weight) in the leaves of Azadirachta indica.
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Fig.l13. Seasonal changes of photosynthetic pigments (mg/g fresh weight) in the leaves of Azadirachta indica.
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Fig.l14. Seasonal changes of photosynthetic pigments (mg/g fresh weight) in the leaves of Azadirachta i n d i c a .
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Fig.l15. Seasonal changes of photosynthetic pigments (mg/g fresh weight) in the leaves of Azadirachta indica.
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Fig.l6. Seasonal changes of photosynthetic pigments (mg/g fresh weight) in the leaves of Azadirachta i n d i c a .
12-14, shows that all the investigated species behave almost in a similar manner
in respect to their contents of the photosynthetic pigments. The amount of
photosynthetic pigments has been found to be the highest in monsoon samples in
all the species irrespective of the level of the pollution.
The Fig. 12-16, show the per cent variation in photosynthetic pigment
between the control and the polluted samples of the investigated species in the
different seasons. Among the different pigments assessed, chlorophyll a shows the
widest variation in T. indica followed by chlorophyll b, while the carotenoids have
been found to vary in a narrow range.
In A. indica and F. bengalensis chlorophyll b and carotenoids show wider
variation than chlorophyll a in comparision to the control sets. In F. religiosa on
the other hand, the carotenoids take the lead to record the widest deviation from that
of control, the chlorophyll b follows the carotenoids, while chlorophyll a records the
least variation compared to control.
Cassia fistula shows that the chlorophyll 'a ' and carotenoids are more
affected than the others, recording the highest variation in the young as well as
in the older leaves, although the level of variation is narrowed down in the
monsoon, where the carotenoids take the lead to record the highest variation among
other pigments.
CARBOHYDRATE IN LEAVES
The data on the total carbohydrate contents in the leaves collected in various
seasons from the species investigated in the present study are summarized in Table-
13. The data reveal that in all the investigated species there has been an increase in
the amount of carbohydrate except in T. indica under the stress of coal-smoke
pollution.
The Fig. 17 indicates the seasonal variation in the amount of carbohydrate
24
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Fig.l 7. Seasonal variation in the contents of carbohydrate (per cent dry weight) in the leaves of various tree species.
in the foliage of different species studied. A glance at the figure clearly shows that
there has been a steady increase in the level of carbohydrate in the leaves of the
polluted population of A. indica, C. fistula and F. bengalensis, from summer to
winter. In T. indica and F. religiosa the peak amount of carbohydrate has been
recorded in the control as well as in the polluted population in monsoon. However,
the variation between the control and the polluted population has been found to be
non-significant in A. indica and T. indica.
The per cent variation in Fig. 20 points out that in C. fistula and F. religiosa
there has been an initial decrease of carbohydrate in the summer foliage followed by
an increase in the other season under the environmental stress. In T. indica on the
other hand, the level of carbohydrate under pollution stress is always lesser than in
control. But in A. indica and F. bengalensis an opposite trend in carbohydrate level
has been recorded, irrespective of the seasonal variation in the climatic conditions.
The seasonal mean given in Fig. 20 indicates that in all the investigated species
there has been an increased percentage of carbohydrate under the stress of coal-
smoke pollution with the exception of T. indica in which a negative effect has been
recorded. The increase happens to be the highest in F. bengalensis (33.01%)
followed by A. indica, C. fistula and F. religiosa (7.70%, 5.53% and 2.06%
respectively). However, the increase in carbohydrate level in C. fistula over the
control has been found to be non-significant.
CARBOHYDRATE IN BARK
The analysis of total carbohydrate in the bark samples of the different species
investigated shows (Table-14) that it increases significantly in the newly formed
bark tissues of A. indica, C. fistula and F. religiosa under pollution stress, while
in the rest of the species the level of carbohydrate undergoes a considerable loss.
The Fig. 18 expresses the seasonal trend of carbohydrate level in the bark
25
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Fig.18. Seasonal variation in the contents of carbohydrate (per cent dry weight) in the bark samples of various tree species.
tissues of the various investigated species It becomes clear that in all the investi-
gated species there has been a gradual increase of carbohydrate from summer to
winter with the single exception of the monsoon samples of the control sets of C.
fistula in which a slight dip in carbohydrate level has been observed. However, in
A. indica, the increase in carbohydrate level has been found to be only marginal
and non-significant.
The per cent variation in carbohydrate level in bark over the control has been
expressed in Fig. 20. It shows that the carbohydrate level falls in T. indica and F.
bengalensis 'in all the seasons with the highest occurring in winter, while in^ . indica
and F. religiosa there has been an increase in the carbohydrate percentage in all
seasons. In C. fistula on the contrary, there has been an initial decrease followed by
subsequent raise in the carbohydrate level under the pollution stress. The positive
variation in carbohydrate level has been found to be the maximum in monsoon
(32.07%) in A. indica and in summer (27.98%) in F. religiosa.
The seasonal mean of carbohydrate in the bark samples of the different species
under taken for the present study is indicated in Fig. 20. A glance at the figure clearly
shows that in A. indica, C. fistula and F. religiosa there has been an increased
percentage of carbohydrate accumulation under pollution stress, while T. indica and
F. bengalensis have experienced severe loss in the level of carbohydrate.
CARBOHYDRATE IN WOOD
The total carbohydrate content in sap wood on analysis shows that, the wood
has more carbohydrate than leaves and bark in a given species. There has been a
decrease in the level of carbohydrate in the sap wood samples of all the investigated
species except in F. bengalensis under pollution stress, in which a significant
increase in the carbohydarte level has been noticed over the control (Table -15).
The seasonal alteration of carbohydrate given in Fig. 19, indicates that the
26
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Fig.l9. Seasonal variation in the contents of carbohydrate (pcrcent dry weight) in the wood samples of various tree species.
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response ofy4. indica, C. fistula tiwA T. /W/'ca is almost similar under pollution stress.
All the above mentioned species have recorded a fall in monsoon in respect of their
carbohydrate content in the sap wood samples. In A. indica and T. indica, the peak
amount of carbohydrate occurred in summer, while in C. fistula the same recorded
in winter. On the other hand, in F. bengalensis there has been an increase in the
amount of carbohydrate from summer to winter. In F. religiosa, the carbohydrate
level has been observed to undergo a marginal and non-significant loss under
pollution stress.
The per cent variation given in Fig. 20 implies that in all the investigated
species there has been a negative variation in carbohydrate percentage under the
influence of coal-smoke pollution with the exception ofF. bengalensis in which there
has been a positive variation in the level of carbohydrate in all the seasons.
The seasonal mean in the level of carbohydrate in the sap wood of the
investigated species is given in Fig. 20. It shows that there has been a significant
decrease in the amount of carbohydrate in A.indica, C. fistula, T. indica and F.
religiosa, upto the extent of 14.65%, 12.29%, 21.50% and 8.53%, respectively
under pollution stress. InF. bengalensis a significant increase of (7.39%) carbohy-
drate occurs under the same environmental condition.
PROTEIN IN LEAVES
The results obtained in the present study on the effect of coal-^moke pollution
on the protein contents in the leaves of the various species investigated are
summarized in Table-16. The data reveal that the amount of protein increases
significantly in all the species investigated in the SO^ enriched environment, except
in T. indica in which the amount of protein undergoes a significant decrease under
the pollution stress.
The seasonal trend of changes in protein in the leaves of the various species
27
ti >
es O U
V M) ee a JS
M) • M V > V9 i Ml "ec e
G V C e u
o u O. 75 o H
£ OS H
® £ B O « m t) M
M V
B
o
E g s
u H 1/3
k-a u
(/3
<S vo oo 1/3 fo' I I
Q Q Q O U U
00 en O m
"ti-cs CJs'
g I •s:
a 1
ro VO C/D (/3
Z Z
Q Q D U U U
<S OO t/3 (S Z ^
I I • — fM Q O O U U U
t--tT 00 r - ^
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5
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Tf vo vo 00 OO
<N 00 csi I I <
f^ f^ ^ pg Q Q Q O Q Q U U U U U U
t- , (N TJ- ON Ov ON o r- in ITl O fS
r-' d vb fS
(S 00 (N ON
VO ( N i n T i - o o ON t s i n VO T i - m VO r - T T 0 0 o
0 0 d 00 0 0 ON
o
t s »—1 VO 0 0
NO 00 o VO >n r- ON O ts VO m 00 o r- ro VO d (N fW (S d fW On r-' On d fS >—1 1—* <
o f<i o c s o o o NO i n CNl ON VO On NO 00 00 o 00 r - 00
ON ( (S •—1 00 ON
U
>1 s: <u
I a o
c l> B to u u B a wi u x:
B o K ca
Q u
Q O
B O 05 « t> CO
U e tB en ti
M
s s B B a as u o kH ^ ^ £ a G u u
Q Q O U
u o 1 1 £ a
cC u
40'
35
30
25
20
15
10 5
0
A.indica. 40'
35
30
25-
20
151
10
5
0
C.fistula.
J M W M W
40-
35
30
25
20
15
10
5
0
T.indica.
KL Jl M W
40
35
30
25
20
15
10
5
0
F.religiosa.
Control Polluted
M W
S - Summer M - Monsoon
40
35
30
25
20
15
10
5
0
F.bengalensis.
LULII S M W
W- Winter
Fig.21. Seasonal alteration in the amount of protein (per cent dry weight) in the leaves of various tree species.
studied are represented through Fig. 21. It becomes clear that in A. indica and F.
religiosa the content of protein gradually falls from summer to winter, with the single
exception of the control sets of A. indica in which the winter foliage has been
observed to have a slight increase. In 7". indica and F. bengalensis, the maximum
amount of protein occurs in monsoon, while in C. fistula the climax is recorded in
winter by following a moderate dip in monsoon. However, the interaction has been
found to be non-significant in all species except F. bengalensis.
The Fig. 24 on per cent variation worked out over the control on the basis of
the contents of protein in the foliage of different species investigated shows that in
T. indica the level of protein falls on the negative side under the coal-smoke
pollution, while in the rest of the species there has been a positive variation in protein
percentage in all the seasons. The maximum decrease has been recorded in winter
(32.44%) in T. indica. The highest increase in the percentage of protein under
pollution stress has been noted in monsoon (59.80%) mA. indica andF. bengalensis
(94.07%), in summer (37.76%) in C. fistula and in winter (47.08%) inF. religiosa.
The seasonal mean given in Fig. 24 manifests that in A. indica, C. fistula, F.
bengalensis and F. religiosa there has been an increased percentage of protein
accumulation under pollution stress (48.68%, 18.87%,51% and 29.01%), while in T.
indica the level of protein undergoes considerable decrease.
PROTEIN IN BARK
The protein content in bark on analysis shows that it undergoes a significant
loss in C. fistula and Ficus spp., under the influence of coal-smoke pollution, while
in the rest of the species this important cellular component builds up positively,
irrespective of the level of pollution in the ambient air (Table-17).
The seasonal variation observed in the protein content of the newly formed
bark tissues in the different investigated species is given in Fig. 22. The figure
28
>
Q U
® e
ce
.2P 'S JS w u i -Sf "5J5 E
e e w
'5 o u a
o H
£ OS H
Z o c« <
e e ON vO 00 in o r- NO 00 ON ee 2 rf lA) o n o\ VO ^ s ON ON 00 r- vo vo m —'
ro (S (N ro m tN 1 (N ri
k V o o TT t- O 00 iri ON On s ON in OS 00 r j
(S vO m <N 00 00 u-i
t> E E 3
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C/3
u 0M
oo C/D C/3 d
I I t
— «N O O P u u u
(N Os Tf GO C/3 d I I I -- fs f
O O P u u o
rs On o\ — (N iTi iri d d d
Q Q Q CJ U U
rr o\ cs C/3 C/5 d IZ IZ
Q Q Q U CJ U
u o u u
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6
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i bo ^
tn •J tj u;
(N c/D C/3 d Z ' I I fSj
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00 On m r- oo o NO r-<s ( S r- NO m ( S o n <s r- Tt m VO m vo ON 00
fO (S ri ( N
o (S <S O o VO o ON irt On O On m (N On
TT Tf o (S <N cs (N — ri
r-CJ 00
CJ
tn C
a s:
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c t) E w il 4J E CO t) JS
c o tn us
Q u
p U
c o m C3
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P D 0 u 1 I
p p O O
-o —. u o 2 I I U
8 -
7
6
J
4
3-
2-1
0
A.indica.
J1 M W
C.fistula.
JL M W
T.indica.
J1 M W
8 -
7
6
5 4 3-
F.religiosa.
Control Polluted
ILLJL M W
S - Summer M - Monsoon
F.bengalensis.
HLJL M W
W- Winter
Fig.22. Seasonal alteration in the amount of protein (per cent dry weight) in the bark samples of various tree species.
reveals that in A. indica and T. iudica there has been a gradual increase in the level
of protein in bark tissues from summer to winter except a slight fall in the level of
protein in the winter samples of the control population of A. indica, while in the rest
of the species the peak amount of protein has been recorded in monsoon, although
the change remains non-significant in A. indica, C. fistula and the Ficus spp.
The per cent variation given in Fig. 24 indicates that there has been a marked
negative variation in the amount of protein in the newly formed bark tissues of C.
fistula and Ficus spp., under the pollution load. The variation widely varies in
summer in C. fistula (25.40%) and F. religiosa (41.67%), and in monsoon in the case
of F. bengalensis (31.53%). The positive variation in the percentage of protein
observed under pollution stress reaches the highest in summer in A. indica (64.60%)
and T. indica (80.20%).
The seasonal mean of protein gievn in Fig. 24 implies that in C. fistula, F.
religiosa and F. bengalensis there has been a significant loss in the amount of protein
in the SO^ enriched environment (22.53%, 36.63% and 26.32%), while in A. indica
(33.59%) and T. indica there has been a considerable gain (52.23%) in this vital
component of cells.
PROTEIN IN WOOD
The data in Table-18 on the effect of coal-smoke pollution on the protein
contents of the sap wood samples analysed point out that ihe amount of protein
significantly increases in A. indica, T. indica and F. religiosa under the influence of
pollution, while in the rest of the species the protein content falls significantly.
The Fig. 23 on seasonal variation of protein in the different investigated
species shows that in A. indica there has been a sharp increase in the amount of
protein from summer to winter. The trend has followed the same course in C. fistula
also, but the increase from summer to winter has been only marginal. In T. indica
29
•a o e
M • M V x: v.
•i: "w E
e e o
'S -t-i O L. D.
e H 00
o: H
V) •w R o u
e £ e o « 2 u A S I/)
B
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U u
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fs VO
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o o o u o u
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a
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o
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Q P O U O U
o
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<N 00 VO OS o CN VO V) ON r- IT) OO
m O r- in T}- OS ON CN) fO CN tsi o i « d d
m rj- CO VO 00 tN rr r- so On CS o r- o m so 'a- t- 00 CO ri CS (si — — — d T—t
U
•2 s.
I en •jj u u;
c <u 6 CO
u JS
§ Vi OS
o u
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c o wi tu V 05
U B cs m u J3
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U U
.s Q Q u u
Q Q U U
T3 — 4) O
^ I
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8' 7
6
5
4
3-2' 1
0
A.imUca.
JLI
8 -
7
6
5
4
3
2
1
0
Cfistula.
M W M W
T.indica,
II DJL M W
gJ 7-
6 <
4 .
F.religiosa.
ILIJI M
El Control • Polluted
W
S - Summer M - Monsoon
7
6 5
4
3 "
2'
F.bengalensis.
i L i d l M
W - Winter
W
Fig.23. Seasonal alteration in the amount of protein (per cent dry weight) in the wood samples of various tree species.
tb < u u
o S
1
s c
c o 8 c o s
s 1
I
a
.2
a
g 1
H
83
e S s o w e '5 e a B B o • mt
•c es > B u h. V
fS ei)
a minor increase of protein has been recorded in monsoon samples compared to other
seasons. On the other hand, in F. hengalensis poWnXtA sets show a gradual increase
of protein from summer to winter but the control population records a slight fall in
protein content in monsoon followed by the peak in winter. However, in religiosa
there has been a gradual but non-2significant decrease in the protein content of sap
wood samples from summer to winter.
The per cent variation worked out over the control and given in Fig. 24 reveals
that it becomes positive under pollution stress in A. indica, T. indica and F.
religiosa. The variation becomes wider in winter iny4. indica {59.26%) and T. indica
(29.63%), and in monsoon in case ofF. religiosa(4S.S7%). However, the negative
variation in protein recorded in the wood samples over the control happens to be the
maximum in monsoon (22.43%) in case of C. fistula and in summer (32.47%) in F.
bengalensis.
The seasonal mean shown in Fig. 24 indicates that there has been a significant
gain in the level ofprotein in^. indica, (44.41%) T. indica {\9.05%) andF. religiosa
(30.60%) under pollution stress, while in C. fistula andF. bengalensis there has been
a considerable loss in protein.
NITROGEN IN LEAVES
The nitrogen content of leaves on analysis shows that all the investigated
species recorded increased amount of nitrogen under coal-smoke pollution,
irrespective of seasonal conditions, with the single exception of Tamarindus indica
in which there is significant decrease in the amount of nitorgen in the polluted
population (Table -19).
The Fig. 25 indicates the seasonal variation in the amount of nitrogen content
in the leaves of the selected species. A glance at the figure indicates that in
Azadirachta indica and Ficus religiosa there has been a fall in the level of nitrogen
30
> in
e8 Q U
CA Z o W5
CA
Sa e e ON ON T}- in r- r-es 2 f l VC ON o ON (S cn o
O o m 00 o ON
ri (N (S
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(S rn d (si cs
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d d o
P Q Q U U U
m o ON f—* m (N O 00 m o r-in CJ ON (S rr r-en r4
VO O VO r- r- rr o m r-O o VO o fO m (S m
T}; ts t- ON t- m (S
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i
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d o wi cs K cn « E ea tfl u
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<2 <2 Q Q u u
p p U U
tii o £ c 1 o ft, O
A.indica. %-
7.
6
5-
3 .
2-1 1 OJ I I
M W
8 -
7 .
6 5-
4-
3 .
2 .
J
0
3.
2.
T.indica.
C.fistula.
M
LULII
M W
M W
F.reUgiosa.
M Control Polluted
J W
S - Summer M - Monsoon
F.bengalensis.
JLIJI M W W - Winter
Fig.25. Seasonal variation in the nitrogen content ( per cent dry weight) in the foliage of various tree species.
from summer to winter irrespective of the pollution level. In Tamarindus indica and
Ficus bengalensis the maximum amount of nitrogen has been recorded in monsoon,
and in winter in case of Cassia fistula.
The Fig. 28 shows the per cent variation of nitrogen in leaves over the control.
A glance on the figure shows that the nitrogen level of the foliage of all the
investigated species is positive under pollution stress except in T. indica in which
a fall in the level of nitrogen has been recorded in all seasons, the variation being
wider in winter, amounting upto 32.47% . In F. religiosa and F. bengalensis the
variation touches the peak in winter (40.11% and 73.57%), whereas in .,4. indica the
variation strikes wider in monsoon (59.77%) and in C. fistula in summer (41.56%).
The mean of the season ( Fig. 28 ) worked out shows that in A. indica, C. fistula, F.
religiosa and F. bengalensis there has been a gain in the nitrogen content under the
pollution stress upto the extent of 40.92%, 20.93%, 24.00% and 36.35% respec-
tively, while in T. indica a considerable loss ( 20.12%) has been recorded.
NITROGEN IN BARK
The data on the effect of coal-smoke pollution on the nitrogen content of the
newly formed bark samples of different species studied show that in A. indica and
T. indica, there has been an increase in the amount of nitrogen to the extent of
significance under the pollution stress. In the rest of the species, on the other hand,
the nitrogen content falls far below the lelvel of control, irrespective of seasonal
conditions (Table -20 ).
The Fig. 26 shows the seasonal variation in the amount of nitrogen in the
newly formed bark samples of the selected species. In A. indica and T. indica there
has been a continuous increase in the amount of nitrogen in the newly formed bark
tissues. While in the rest of the species the nitrogen content showed its peak in the
monsoon samples. However, the interseasonal variation in A. indica and the
31
>
Ji
tn 4-t R ft u
JES
O £ e o es 2 ti a S
c
Z O c« <
CA hi V E E s
H NM IZ5
U u A. </3
ro —' ( S < s o o q d d d
Q Q D U U O
U
a I
§
t-O C/D C/3 d ^
f^ ^ ^ (N <N o o o o o o
I
Q Q Q U O O
P Q Q O O U
i r^ i r . O —< (N (S o o q d d d I < I « rJ f
O Q G U O O
U U CJ
Q
a
c
c
o
a
00 o d I
m tN O o d o i I
O D D U U O
vO vO _ o r- oo 0 0 o ON i n m o iri 0 0 VO T f vo vO f S
d d d d d d d d d d
r - r - VO r - CN r o vo >—1 Tj- o o r - CS i n
O) vq vq t- 0\ CN I S m
d d d d d d d d d d
, o o > i n VO VO m o 'iS- o x f VO o\
t - - VO 00 T l - \o ts d d d d d d d d d d
VO » r- m 00 TT «n >n VO m o n 00 o Ti" ( S
TJ- n Tf <N f o
d d d d d d d d d d
U
R
a s:
o
c u E w CO u U E to VI « J3
e o to 03 4>
Q U
Q U
c o w (S u V, V E CS M u
u u E E « eo
£
D D U U
Q Q O O
u o
= I
6
A.indica. C.fistula. 1 6-
1.4-1.2
1.0-
0.8-
0.6
0.4
0.2
0 J1
1.6-
1.4-
1.2 1.0-
0.8-
0 . 6 -
0 .4 -
0.2 J
0 JL M W M W
T.indica. 1.6-
1.4-
1.2
1.0-
0.8-
0.6-
0 . 4 -
0.2 J
0 JUU M w
1.6
1.4
1.2
I . O
0.8
0.6
0.4
0.2
F.religiosa.
LUUI M W
Control Polluted
1.6'
1.4
1.2
1.0 0,8 0.6 0.4
0.2
0
S - Summer M - Monsoon
F.bengalensis.
JLILII M W W - Winter
Fig.26. Seasonal variation in the nitrogen content ( per cent dry weight) in the bark samples of various tree species.
summer and winter samples ofF. hengalensis, do not show any significant difference
under normal condition.
The Fig. 28 on per cent variation shows that a low level of nitrogen has been
found in the newly formed bark tissues of C. fistula and F/cw5spp. under pollution
stress. However in the rest of the species the level of nitrogen has been found higher
in control. The highest percentage of variation has been observed in the summer
samples (Young leaves) of A. indica (63.34%) and in the winter samples (Older
leaves) of T. indica (91.31%).
The mean of the season noted in respect to its nitrogen content shows that in
C. fistula, F. religiosa and F. bengalensis there is an overall loss of nitrogen under
the pollution stress, while in A. indica and T. indica a considerable increase in
nitrogen content has been recorded.
NITROGEN IN WOOD
The data summarized in Table- 21 show that in A. indica, T. indica and F.
religiosa there has been an increase in the amount of nitrogen in the sap wood
samples of the polluted populations, while in the rest of the species, an opposite
condition prevails under the stress of coal-smoke pollution.
The Fig. 27 elucidates the seasonal variation in the amount of nitrogen in the
sap wood samples of the selected species. A glance at the figure clearly shows that
in F. religiosa there is a continuous fall in the level oi nitrogen both in the control
as well as in treated, while the rest records a more or less gradual increase from
summer to winter with a sharp fall in case of F. bengalensis and a non-significant
increase in case of T. indica in monsoon.
The Fig. 28 on per cent variation shows that,under the stress of coal-smoke
pollution A. indica, T. indica and F. religiosa show positive variation in nitrogen
percentage in the sap wood samples. The variation is wider in the younger leaves
32
u >
lo es Q U
-a o e ^ u us
x: M • M
T5 e w u V a
B B e w B V bfi e • M B Is
fS JU s 03 H
® s B e ee 2 ea
u B
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c« t. E E s
H
1/3 U Nw u u eu, C<5
lO vO — (N (N o o o o b b I I I « r Q Q O U U U
OS
u
S c
o
Q N
cn fn — <N f^ o o o b b b I I I -i fN) m
Q Q O U U O
— OS OS O O O b b b I I I — fs
Q Q O U O U
t-- »/-> o o o b b b
O O P U U U
U O CJ
t»3
Q tc
a
?5 O s:
c
I
cs o
v. t^ B O
— o ^ o o p b b b • I ' — <s r
Q O G U U U
OJ ro 0 < 0\ CO TT in DO <r) OS o m 00 VO m (S fS ts (S fS ts b b b b b b b b b b
OS 00 (N cn OS o 00 m O o 00 rr 00 (N
00 r- ts b b b b b b b b b b
ro cs so so o en o lyo so so F-H so ts (N r-i CN b b b b b b b b b b
1 so in OS o o so O 00 m o in cs m (N (S rs rsj
b b b b b b b b b
U
s:
I <1
o
C u E CO £
E cd ir. u
i eg U CO k' <S Q o
O u
c o CA ca i> cn <u e C3 VI Hi
C C o <u B 6 SB <0 U U
,o Q O O O
Q D U O
•S o W u ^ c s a
fC O
1 6
1 4
1 2
1 0 0 8 0 6 -
0 4 '
0 2
0
A.indica. 1 6 -
1 4 1 2
1 01 0 8
0 6 1
0 4
0 2
0
C.fistula.
JLM M W M W
1 6
1 4
1 2
1 0
0 8
0 6
0 4
02
0
T.indica.
II II il M W
1 6
1 4
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1 0
0 8
0 6
0 4
0 2
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F.religiosa.
Il II II M
Q Control • Polluted
W
1 6-
1 4 1 2
1 0 0 8 0 6 '
0 4 -
0 2
0
S - Summer M - Monsoon
F.bengalensis.
ILHLL M W
W - Wuiter
Fig.27. Seasonal variation in the nitrogen content ( per cent dry weight) in the wood samples of various tree species.
< til
I u H
c 8 (A G
I
I
I
S I I c c
i •s, a
I
s 1 e •I 2
I •
g e o V e Sd e
• mm e e
es •c es > e u u V Pm 00 <s w
S o 2 T o c o 7 •= s T o o o o- o
(Summer foliage) in A. indica (31.14%), and T. indica and F. religiosa in
the older leaves i.e. winter foliage . On the other hand, in C. fistula and F.
bengalensis the variation falls on the negative side in all the seasons under
pollution stress, the highest variation occurring in monsoon.
The mean of the seasons ( Fig. 28) worked out in respect to its nitrogen per
cent indicates that in A. indica, T. indica and F. religiosa there has been a
significant gain in nitrogen in the sap wood samples under the influence of coal-
smoke pollution, while in the rest (C. fistula and F. bengalensis ) there has been
a fall in the nitrogen level under the coal-smoke pollution.
PHOSPHORUS IN LEAVES
The data (Table-22) on the effect of coal-smoke pollution on the phosphorus
content of the leaves show that in Ficus spp. there has been an increase in the amount
of phosphorus in polluted populations, while in A. indica and T. indica the level
of phosphorus recorded a significant fall compared to control. However, in C.
fistula the level of phosphorus remains almost at the same level irrespective of the
level of pollution.
The Fig. 29 shows that in A. indica, C. fistula and in Ficus spp., there has
been a continuous fall in the level of phosphorus from summer to winter except in
the monsoon samples of the control sets of C. fistula in which a significant raise has
been recorded. In all the investigated species the peak concentrations of phosphorus
has touched in summer (i.e. young foliage) except in T. indica which has a monsoon
peak.
The Fig. 32 on per cent variation worked out on phosphorus content in the
foliage of the investigated species in various seasons shows that in Ficus there
has been a positive variation in the phosphorus percentage under the pollution stress,
with the maximum in summer in the case of F. religiosa (19.04%) and in the
33
> Wi
es O U
O S e o es 2 2 C»
u e
Z o <
»5 u V E E a
H
en
u ft*
O f^ •*}• <N rf- fO O O O d d d I I I
— IS O O P o u u
u
y c
Q N
Tl-— <N <N o o o d d d I I I — fM
P O P u u u
vo o — — o o o d d d I I I — (M
P P P u u o
tA) 3 : S o P p
d d d
P P P U O U
O
a
53 O c
"I Q S
o
r^ tn oc O O o d d d
P P P U U U
< s ( S 0 0 o vo Os ON t - Os i n 0 0 o o m o I T l t s ( N CS ( S ( N m
d d d d d d d d d d
' O VT) r—1 o o o o 00 00 m o t s 00 \D ON
1—1 (S (S (S c s a en CS d d d d d d d d d d
00 i n T T o o ON c s O 0 0 0 0 I T l
c s CO c s c s en
d d d d d d d d d d
<7\ c s vo vo TJ- r f o o o 0 0 vo m c s r r ON i T ) o c s r - c s
r r m c s c s f S
d d d d d d d d d d
O
•2 s:
a c «o O
B U E «
V E w u
B O <n cs V in
D u
P U
a o m ts ID m u e cd v> V
B B
E E CO e3
D O U <J
P P O U
•o — u o I ^ I c5 a. U
0 g 0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
A.indica.
M W
0 8 0 7J
0 6
0 5
0 4 0 3 .
0 2.
0 1
0
0 8-
0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
C.fistula.
iLlUl S M W
T.indica,
lUUl S M W
0 8 '
0 7. 0 6
0 5 l
0 4
0 3.
0 2.
0 1
0
F.religiosa.
M
Control Polluted
H w
0 8 -
0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
S - Summer M - Monsoon
F .bengaJensis.
M W
W - Winter
Fig.29. Seasonal alteration in the phosphorus content (percent dry weight) in the leaves of various tree species.
winter in the case of F. bengalemis (24.16%). In C. fistula and in T. indica,
on the other hand, there has been a positive variation in the phosphorus percentage
in the young foliage of summer samples, followed by a sharp fall in the negative
side in the subsequent seasons. The negative variation of phosphorus in the
foliage has attained the highest under pollution stress in winter in case of C. fistula
and T. indica, while in A. indica, there has been a wider variation (Negetive)
of phosphorus in the foliage in summer and monsoon followed by a significant gain
in winter under the pollution stress.
The seasonal mean (Fig. 32) in respect of phosphorus content in the different
species investigated has shown that all the investigated species have recorded a
higher percentage of phosphorus content from the normal, under the pollution stress
with the single exception of T. indica in which a depletion of 5.92% of phosphorus
occurs. The maximum gain has been recorded in the Ficus spp. and the minimum
in A. indica.
PHOSPHORUS IN BARK
The phosphorus content in the newly formed bark on analysis has revealed that
it increases to a level of significance in A. indica and F. religiosa under the coal-
smoke pollution, while in the rest of the species there has been a significant depletion
of this vital element (Table-23)
The Fig. 30 indicates the seasonal variation in phosphorus content in the bark
of the investigated species. It becomes obvious that in C. fistula, T. indica and
F. religiosa, there has been a continuous increase of phosphorus from summer to
winter, irrespective of the pollution level, while in A. indica and F. bengalensis,
the trend is slightly disturbed by the peak concentration attaining in monsoon in the
former and a sharp and significant fall taking place in monsoon in the case of the
latter.
3A
iS e« A
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IT)
CB Q u
® £ e e « 2 U A
I. W e
.e •Sf 'Z
•o e V V u V
le S h o £ a n O X a o •*-> B S O E <
*
z e« H
CM Z o
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E E s
H CM
U U
f^ vo VO O O O o o o
o o o I
O D Q O U U
o
VO fO o
o
VO O d
cs o
U
Q o s :
- s : 0
1
VO —' 00 o — o o o o d o d
Q D Q U U U
Tf r-.m o o o o o o
o I
o I — fM
Q P O U U U
T i - r - - oo o o o o o o d d d I I I <s
P O O u o u
00 o
On o r-
o
r-i/-) o
vO o d
00 o o O o
d
rr o
U U U
Q
<3
s 1 1
I Sd ^
a o
C/3 c/l t/5 I I I — <N m
P P P u u o
vo o
cs 00 o
d
vO c s OS t - - r - i/^ t s t -en m r l - 0 0 T r 0 0 OS 0 0 0 0 O o o o o
d d d d d d d d d d
^ 1 CM < s t - - 0 0 VO VO >/-) 00 o VO m O o o o o o o
d d d d d d d d d d
in VO O O
U
•2 c
c o <<
o
e u E es V
U E m <« u JS
B s ta u tn k. <2 Q u
I
P u
i CO o tn 0 1 u jC
C B u u 6 S e3 <S u u
£ £ Q Q U U I •
P P u o
I £ 3 B 1 cS
0, u
040
030
Ol'i
020-
015-1
010 0 0 5
0
A.indica.
• I l l • • M W
040
035H 030
025 I
020
015J
oloj 005
0
C.fistula.
JL { M W
040 035 H 030 0^1 0^ 015.J 010,
005 0
TAndica.
ll M W
040-1 035-030 0^1 OJO 015. 010.
0 05
0
F.religiosa.
ll ll II S M w
B Contro] • Polluted
040-035 030
OJO 015 010 005 0
F.bengalensis.
S - Summer M - Monsoon
JLLII M W
W - Wmter
Fig.30. Seasonal alteration in the phosphorus content (per cent dry weight) in the bark samples of various tree species.
The Fig. 32 on per cent variation worked out on the basis of phosphorus
content over the control has shown that in case of C. fistula, T. indica and F.
bengalensis, there has been a fall in the level of phosphorus irrespective of the
prevailing seasonal conditions with the maximum occurring in monsoon in C. fistula
and T. indica, and in winter in the case of F. bengalensis. However in the rest of
the species, there has been a positive variation in the phosphorus content against
pollution stress. The variation in phosphorus content has occurred to the maximum
in summer in case of A. indica (33.33%) and in monsoon in case of F. religiosa
(37.50%)
The Fig. 32 on mean of the seasons shows that there has been a significant
depletion of phosphorus in C. fistula, T. indica and F. bengalensis under pollution
stress (21.75%, 20.18% and 21.95%), while in A. indica and F. religiosa there
has been a significant gain of phosphorus.
PHOSPHORUS IN WOOD
The data given in Table-24 on the effect of coal-smoke pollution on phospho-
rus content of the sap wood samples of the various investigated species show that
in A. indica and F. bengalensis there has been a significant increase of phosphorus
under the pollution stress, while in rest of the species the level of phosphorus shows
a significant fall.
The f ig. 31 on seasonal variation of phosphorus in the sap wood samples of
the different investigated species shows that there has been an increase in the level
of phosphorus from summer to winter in all the species except in A. indica and the
polluted sets of C. fistula, in which the peak touches in monsoon in former and a
gradual fall in the latter.
The Fig. 32 on per cent variation worked out on phosphorus content of sap
wood samples indicates that C. fistula, T. indica and F. religiosa experience a
35
>
VI ee Q
.e
® £ e o CB 2 V A 2 c»
k 4-1 e
O <
c« b. w E E s 03
H
C/D
u u CL,
t^ — o o — o o o d d d I I I ^ r) r
D P Q (J U U
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o
00 o o o GOO rr 00 o o o o o o
o o o I
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o o
Q O O U U O
Q Q Q U U U
o O C) d d d
Q Q Q (J O U
O U a. u
Q ^ tn CS
U
s
c
I « o §
^ o o o o o p d d d • I ' — r^
O Q Q C U CJ
t-- 0\ 00 00 r- m 00 o\ m o V-) o IT)
fN fS (N fS (S (N ( S ( N
d d d d d d d d d d
VO r o t N fM CO f o 0 0 0 0 Ov 0 0 r r m
CS t s t s c s
d d d d d d d d d d
o 0 0 0 0 en > 1 vo fn iri o\ VO O CS CS CS CS CS CN CS CS
d d d d d d d d d d
'SI- Ti- oo 0 0 ITl o • 1—c 0 0 es m VO CS CS SO i r i 0\
CS CS CS <—' CS 1—< CS
d d d d d d d d d d
U
s:
a K
-O
k:
c Hi E « (L> 4> a ea <f> u
B O tn 03 V
Q u
Q U
c o to CB U « U B es CO v X!
c a u u e B CS es U V i i! U Ur
D O u u
Q Q U U
•V V
3 "K O Oh
B 0 u 1
O
0 8
0 7, 0 6
0 5 0 4'
03J 0 2.
0 1
A.indica.
LUHL
0 8-
0 7 ,
0 6
0 5 -
0 4
0 3 .
0 2.
0 1
C.fistula.
M W M W
0 8-1
0 7 0 6
0 5 1
0 4
0 3 0 2,
0 1
T.indica.
M W
0 8 -
0 7 ,
0 6
OS--0 4
0 3 0 2,
0 1
F.religiosa.
JLILII M
B Control • Polluted
W
0 8 0 7
0 6
0 5
0 4
S - Summer M - Monsoon
F.bengaJensis.
S M W
W - Wmter
Fig.31. Seasonal alteration in the phosphorus content (per cent dry weight) in the wood samples of various tree species.
b. < U iJ
FT"'
§ g
c
I a c
c o 2
0 1
60 §
a JS .So
e B 8
o jd a M o JS a s B .2
•E A > •M B « U u Pm fo M)
a
§
5
•5 g 1 •
o o + *
o u- O O o n -r £ S ? o o o
fall in phosphorus content under the pollution stress. The fall attained the highest
level in the winter season in C. fistula and F. religiosa, while in T. indica, the
summer samples has recorded the highest variations (28.07%). However, the rest of
the species have exhibited positive variation under the pollution stress. The
variation has grown wider in winter in case of A. indica (30.38%) and in the summer
in case of F. bengalensis (30.89%).
The seasonal mean of phosphorus level shown in Fig -32 indicates that C.
fistula, T. indica and F. religiosa have been facing considerable depletion in their
phosphorus content under pollution stress (15.50%, 21.78% and 23.41% respec-
tively), while A. indica and F. bengalensis have been gaining considerable amount.
POTASSIUM IN LEAVES
The data in Table-25 show that the potassium content increases to a level of
significance in Tamarindus indica and undergoes a significant loss in F. bengalensis
and F. religiosa, while in the rest it remains almost at the same level irrespective of
pollution level.
The Fig. 33 shows the variation in potassium level in the different seasons. In
A. indica and C. fistula, the potassium level has touched the peak in summer in the
polluted populations, while in the control the same recorded in monsoon samples of
the leaves. In T. indica and F. religiosa there has been a fall in the content of
potassium from summer to monsoon and then a slight increase in winter. But in F.
bengalensis there has been a continuous fall from summer to winter in both the
polluted and control populations. In both the species ofFicus the highest amount
of potassium has been recorded in the summer samples of leaves irrespective of
pollution level.
The Fig. 36 on per cent variation indicates that both Ficus religiosa and F.
bengalensis show a negative variation in the level of potassium under pollution stress
36
>
08 Q U
JS
® £ e o M (A S t» 2
L. s
CA Z o c«
CO 4t E E 9
H O)
05 M u 0k
<N (N lyi m iz; d d
Q Q Q U O U
O
t> t-
r - ' d
00 VO
00 On
u
Q u ^ R
-s: u cs
a N
— 00 O ON
on —; p ^ d d
Q Q O (J O (J
^ oo r-IT) 00 oo o o o d d d
P Q Q CJ U U
(N On 'a-O O d d d
Q Q Q U U U
VO
—I o
ON <N
m f—4 r-d
fO VO
o f—< ON d
iri 00 00
— — o NO d
r-00 d
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t-NO 00
o ts o o —
CJ U U
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Q o R
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I
§ O
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Q Q Q U U U
0 r x ON rr NO NO 00 10 r-
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NO 00 cs 00 00 ON in tr, r- NO NO m 00 cs NO
I—1 d I-H
<N rf ON Tl- CS m t-- cs 00 r- cs • 00 0\ NO
r- 00 00 NO NO NO vr. r-d d d d —
m m tN oo (S ^ —'
' •«» to s:
s: -c
o k:
c u B cs w t> E CO tn U
§ w cs u
D u I rj
Q CJ
c o en ea u w e CS en U
C C I I cs cS (U V k- u u u <2 ^ D D u u
D Q U U
•o —. u o
11 Cu U
4 On 3 5
3 0
2 5
20
1 5
1.0
0.5
0
A.indica. 4 0 3 5 -
3 0
2 5 2.0
1 5 -
1 0.
0 5
0
C.fistula.
M W JULJI
S M W
4 0 -
3 .5
3 .0
2 5 1
2.0
1.5-
1.0.
0.5
0
T.indica.
JLIII M W
4 0
3 5
3 0
2 5
2 0
1 5
1 0
0 5
0
F.religiosa.
M B Control • Polluted
W
4.0
3 .5
3.0
2 5
2 0
1 5
I 0
0 5
0
S - Summer M - Monsoon
F.bengalensis
I M W
W - Wmter
Fig.33. Seasonal variation in the potassium content (per cent dry weight) in the leaves of various tree species.
in all the seasons. But C. fistula records a non-significant negative variation of
potassium in monsoon and significant variation in winter season, after experiencing
a significant positive variation in summer. In all the three above species maximum
variation of potassium has been recorded in winter. In T. indica, on the other hand,
there has been a positive variation in the level of potassium under the pollution stress,
while in A. indica the variation falls on the negative side in monsoon and positive in
the other seasons.
The mean of potassum level in leaves indicates that T. indica accumulates the
highest amount of potassium in leaves under pollution stress (22.47%) followed by
C. fistula (4.62%) and A. indica(2.60%), while in Ficus spp. there is a significant
depletion of potassium.
POTASSIUM IN BARK
The data in Table-26 show that the potassium content falls to a level of
significance in the newly formed bark samples of A. indica and Ficus spp., under the
stress of coal-smoke pollution, while in T. indica it shows a significant increase
over to its control under the same environmental conditon. However, in C. fistula
the content of potassium remains at the same level irrespective of pollution level.
A glance at the Fig. 34 clearly indicates that A. indica and F. bengalensis
behave more or less in a similar manner with respect to its level of potassium in the
newly formed bark samples under pollution stress. In both the species the highest
amount of potassium recorded in monsoon samples of the bark both in the control as
well as in the polluted samples. In C. fistula there is a significant increase in the
amount of potassium from summer to monsoon and then there is a significant fall in
winter. In T. indica a gradual increase of potassium from summer to winter has been
recorded. But in F. religiosa there is a significant fall in the amount of potassium
from summer to monsoon and then there is a slight increase in winter.
37
ti >
OS O u
k M iC V
® £ e o oj es
e
M) •mm
•o
9i W V D.
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(A ee o a
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C/3 Eii] u
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—' ro — m r-o o o d d d I I I
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o C/D (/3 o z
Q Q Q U U U
D Q Q U U U
Q Q Q U U U
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s: 53
14)
U u ;
(N 00 — li?
O o P d o d
Q Q Q U U U
V-) <N (N fN 00 <N VO r- c VO 00 ON •ri cn in Ti- 00 d d d d d d d d d d
c s r-H o ,—1 00 o r -VO T-M VO I T l «N (S VO Ov iri d d d d d d d d d d
fo Ov 00 VO in <N VO •rr 00 < s CM rri r -
r t VO 00 VO d d d d d d d d d d
00 r- (N ON 00 VO vO VO o f S VO f—1 ON fn VO o m in (N 00 o I T ) lA) d d d d d d d —' d d
U
tn s:
I o to o
c u E eo «
U E C8 trt U M
a o tn cs u
D U I
Q U
B O <n u E CD CO u
g g e 6 a cs
.5 .P Q D U U
Q Q O U
T3 — U O = I
I I a. u
A.indica. C.fistula. I 1 4 .
) 2 1 0 '
0 8 -
0 6 .
0 4 .
02
0 JLLML S M W
] 6 -
1 4
1 2 l
1 0
0 8
0 6
0 4
0 2
0 -
I 6-1 4 -
1 2 '
1 0 '
0 8 '
0 6 .
0 4 .
0 2
0 •
JLI
JLM S M W
T.tndica.
M W
1 6
1 4
1 2
1 0
0 8
0 6
0 4
0 2
0
F.religiosa.
I S M
El Control • Polluted
W
1 6
1 4
1 2 1 0
S - Summer M - Monsoon
F.bengalensis
M W W - Wmtcr
Fig.34. Seasonal variation in the potassium content (per cent dry weight) in the bark samples of various tree species.
The Fig. 36 shows the per cent variation in the potassium content of newly
formed bark. It clearly indicates that in A. indica and Ficus spp. the variation falls
on the negetive side in all the seasons. The wider variation occurring in monsoon
in the case of F. religiosa and in the winter in F. bengalensis and A. indica. In T.
indica, on the other hand, the level of potassium shows positive variation under
pollution stress with the maximum attaining in monsoon (29.50%). In this respect
C. fistula showed a unique behaviour compared to others showing positive variaton
of potassium in summer and winter samples under pollution stress (43.86% and
11.95% respectively), but in monsoon season a significant fall in the negative side
has been observed (27.55%).
The seasonal mean of potassium content (See fig. 36) indicates that in A.
indica and Ficus spp. there has been a significant depletion of potassium under
pollution stress (20.72%, 24.60% and 11%), while in T. indica an opposite trend has
been noted. However in 'C. fistula the depletion of potassium recorded under
pollution stress has been marginal and non significant.
POTASSIUM IN WOOD
The potassium content of the sap wood samples of different species on
analysis shows that in A. indica, C. fistula and F. bengalensis, there is an increased
amount of potassium under pollution stress, while in the others the potassium level
undergoes a severe loss (Table -27 ).
The Fig. 35 on seasonal variation of potassium in the sap wood samples of
different species shows that indica and C. fistula behave almost in a similar manner
under the pollution stress. In both the species peak concentration of potassium
touches in monsoon except in the polluted population of C. fistula in which peak
touches in summer. In T. indica peak concentration of potassium recorded in summer
samples of the wood of polluted sets, while in the control sets the same has been
38
> a
Vl et Q U
® £ e e
e:
CZ3
iz: O
w V 6 g s
c «
H
D w On
<N O C/D C/5 d
Q Q Q O U U
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(S ^ t^ —' <N — o o o
d d d
Q Q Q U U O
ir, \o iT, —' (N (N o o o
d d d
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o ff) — o o
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-- m O Q Q U U U
u u
a
S c
i c
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§ o
o
00 (N vo X ^ ^ O O p d d o ' I I <
— n r^ Q Q O U U U
VO rr t-- fM o ( S 00 p—t o m lA) o ID o ( S <N <N
d d d d d d d d d d
00 CN o (M 00 o ON o VO ( N ON 00 v£) o CS <N CS CS r-d d d d d d d d d d
CS CS 00 On lA) o 00 00 r - CS ro in
CS CNl cs m m CS r o CS d d d d d d d d d d
o On o r- ,—1 00 NO 1/1 On m lo On o o 00 VD CS CS m CS CS m CS d d d d d d d d d d
U
>1 to c
I
B u e <0 u u E es
O
§ cn ca V tn
o u
Q U
1 CS u V E a u .c
B B I I es es U u
D D u u I I —• n
Q O U U
•H £ — B g 5 CLh U
0 8
0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
A.indica.
JL M W
0 8
0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
C.fistula.
M W
0 8-
0 7 .
0 6
0 5 -
0 4
0 3 .
0 2 .
0 1
0
TAndica.
I I M W
0 8 0 7 .
06
0 5-
0 4
0 3 .
0 2.
0 1
0
F.religiosa.
J S M
Control Polluted
W
0 8'
0 7 0 6
0 5 0 4 0 3 0 2.
0 1
0
F.bengalensis.
S - Summer M - Monsoon
1 M W W - Winter
Fig.35. Seasonal variation in the potassium content (per cent dry weight) in the wood samples of various tree species.
(b <
I
0 c 1
I .So
a
5 1
I «
B S C e w
tn tn es o a
B .2 '•C e« •c
es >
B V u h £ NO ei)
I .g
a
2
•g g
w
g O I - vo o O o — " T o o
recorded in monsoon. However in both the sets the amount of potassium happens
to be the least in winter. The behaviour of Ficus spp. has also been noted to be almost
similar except there is an increase in the contents of potassium in the polluted
population of the winter samples of F. bengalensis, whereas F. religiosa has
recorded a decrease in the content of potassium in winter samples of the polluted
populations.
The Fig. 36 on per cent variation of potassium content in sap wood samples
shows that in F. religiosa there is a significant fall in the negative side in the level
of potassium from summer to winter, with the maximum occurring in monsoon
(68.44%). While in T. indica there is a positive variation in the potassium content
in the summer samples of the sap wood (49.80%). However, in the other seasons
there has been a negative variation.
In F. bengalensis the positive variation has been observed in monsoon and
winter season attaining the maximum in winter (47.40%). In A. indica and C. fistula
the variation goes on the postive side in the amount of potassium in all seasons. The
highest percentage of variation has been observed in winter in C. fistula (32.23%)
and in summer (16.46%) mA. indica.
The seasonal mean (Fig. 36) of potassium content in wood samples shows
that T. indica and F. religiosa experience a significant depletion of potassium
under pollution stress (8.73%, 11.25%), while the others giving a significant
increase.
SODIUM IN LEAVES
The present study on the effect of coal-smoke pollution on the sodium content
in the leaves of the various investigated species reveals that in A. indica, C. fistula
and T. indica, there has been a significant increase in the level of sodium under
pollution stress, while in the rest of the species, the level of sodium undergoes a
significant loss (Table-28).
39
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x:
® £ e o es 2
UD .S is
fc. a
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U) • M
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m O C/3 C/3 6
Q P Q D U O
o Ov O
O cs
CJ
I
r- (N cs o o o d d d
P Q Q O U a
r- t^ o <s - r q o o d d d
P P P U U U
(S o iri cT)
d z I • • ^ r>j n
P P P u o u
o r-ro d
O in ro O ts
o 00
o <s
o tN d
U U U
55 Or
tS
55
C
s: 55
I
a O .Ss (U V. S
— 00 IT s O o P d d <=>
P P P U O U
o o O o O o o o O o o rj- (—1 TJ- m 0 0 r-
( S ( N ts rn d d d d d d d d d d
o O o o o o o o o VD m o 0 0 o t s t s CO ( S < s ( S o
d d d d d d d d d d
o o O o o O o o o O 00 CO VO m o <N r}- ON
rj- in TJ- Tl-d d d d d d d d d d
o ON
O tT ( S
O
s:
I -C)
Ss o
c il E eg 0) 4> E « V) w
B O tn «o u
G o
p U
(3 O tn 03 10 tn « g 03 in 4)
JS
es B B V 0) E E es CS V V ti w ^ ll o o o D o u
P P O U
« o
0 8 "
0 7
0 6
0 5
0 4
0 3
0 2 0 1
0
A.indica. 0 8
0 7
0 6
0 5
0 4
0 3
0 2 0 1
0
C.fistula.
I M W M W
0 8-
0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
T.indica.
JL M W
0 8 "
0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
F.religiosa.
I B Control • Polluted
M
0 8-
0 7
0 6
0 5
0 4
0 3 l
0 2
0 1
0
F.bengalensis.
W S - Summer M - Monsoon
Jl S M W
W - Winter
Fig.37. Seasonal alteration in the amount of sodium ( mg/gm dry weight) in the leaves of various tree species.
The Fig. 37 indicates the seasonal variation in the level of sodium in
different species. It clearly shove's that in all the investigated species, the peak level
of sodium has been recorded in monsoon samples, irrespective of the pollution
level, although the interseasonal variation occurring in A. indica and F. religiosa
has been found to be non-significant under pollution stress.
The Fig. 40 on per cent variation worked out on the basis of the contents of
sodium analysed in the foliage of various species studied indicates that in A. indica,
C. fistula and T. indica the per cent variation is positive under pollution stress with
the maximum occurring in the foliage of monsoon (23.68%) in^^. indica, in winter
(47.82%) in C. fistula and in summer (33.33%) in T. indica. In the Ficus spp. the
per cent variation happens to be negative indicating the depleted level of sodium
irrespective of the seasonal variation in the physical factors of the site.
The seasonal mean given in Fig. 40 points out that the Ficus spp. have
experienced considerable depletion of sodium to the extent of 20.58% to 27.27% in
the different seasons under pollution stress. In A. indica, C. fistula and T. indica,
on the other hand, the variation has been on positive side to the extent of 17.64%,
20.58% and 17.85% respectively under the same environmental condition.
SODIUM IN BARK
The observation recorded on the contents of sodium in the newly formed bark
samples of the various investigated species indicates that the amount of sodium
increases to a level of significance in the bark tissues of A. indica, C. fistula and F. ^
religiosa under pollution stress, while in the rest of the species, there has been a
significant depletion (Table -29).
The Fig. 38 indicate the seasonal trend of sodium balance in the different
investigated species. In A. indica and F. religiosa, there has been a raise in the level
of sodium over the control under the pollution stress, the peak striking at the winter.
117
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Q U
B O « 2 Oi 08
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On O r-<N VO O O O o d d
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6 ^
Q Q D O U U
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a
55
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o
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P D Q U U U
o o o o o o o o O o ON Tf o NO CN O
I T i NO
d d d d d d d d d d
o o o o o o o o o o ON 00 ON 00 m NO OS Tf NO NO Tj- IT) ITl <s d d d d d d d d d d
o o o o O o r- o O O ON ON iri m o m Tt; NO
d d d d d d d d d d
o o o o o o o o o o NO o NO fS ITi 00
m in <-1 <N cs cs CO d d d d d d d d d d
U
>1 <ri s:
I C
g E
eo 4)
U E <a 01 IL>
C o « eo U
0 CJ 1 cs p
u
c o tn ea u tn u E
V JC
B C V V
E E a to u 4>
£ . 2
D Q u u
p p U O
•S o
2 c5 fC u
0 8 "
0 7
0 6
0 5
0 4
0
0 2
0 1
0
A.imltca. 0 r 0 7
0 6
0 5
0 4
0 3 1
0 2
0 1
0
Cfistula.
M W M W
0 g-* 0 7
0 6
0 5
0 4
0 3 1
0 2
0 1
0
T.indica.
Jl M W
0 g 0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
F.religiosa,
1 B Control • Polluted
M
0 8 0 7
0 6
0 5
0 4
0 3
0 2 0 1
0
F.bengalensis.
W
S - Summer M - Monsoon
I S M W
W - Winter
Fig.38. Seasonal alteration in the amount of sodium ( mg/gm dry weight) in the bark samples of various tree species.
In C. fistula the trend of variation has followed the line but the peak occurs in
monsoon. In T. indica, the level of sodium increased from summer to winter but the
pollution stress has kept the sodium level lower than the control. In F. bengalensis
also the level of sodium has remained low indicating the severe depletion of this light
metal under the pollution stress with the maximum amount of depletion taking place
in winter.
The per cent variation worked out on the basis of sodium content in the bark
tissues of the various investigated species is indicated in Fig. 40. The figure implies
that in T. indica and F. bengalensis there has been a negative but non-significant
variation in the level of sodium under pollution stress, while in the other species the
variation has been positive and significant. The variation becomes wider in summer
in case of A. indica (53.33%) and F. religiosa (45.83%) and in monsoon in the case
of C. fistula (S2.05%).
The seasonal mean of sodium given in Fig. 40 reveals that T. indica and F.
bengalensis have experienced significant depletion in the sodium content in the
newly formed bark tissues under pollution stress (22.50% and 36.17%), while in A.
indica, C. fistula and F. religiosa there has been a considerable gain (35.89%,
56.09% and 43.75%).
SODIUM IN WOOD
The sodium content, in the sap wood samples, on analysis shows that it falls
to a level of significance in all the investigated species under pollution stress with the
single exception of T. indica in which the sodium level significantly goes up (Table
-30).
The Fig. 39 on seasonal variation shows that in Ficus spp., there has been a t
gradual increase in the amount of sodium from summer to winter, while in T. indica
it falls in winter after attaining the maximum in monsoon. In C. fistula, the sodium
41
> CI
*n cs O U
V JS
•s = c o « 2 es
v> e
1/5
z o in <
c« u E E s
H
C/5 u U Ed 0.
0\ TT <N — ri Tf o o o o d d I I I —• fM Q O O O U U
CJ
Q o c;
-c u H Q N
fS O C/3 C/3 d z
p p o u o u
00 <
O C/3 C/5 d Z Z
I I I ri P P P u o u
t^ o o o o o o d d
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O O U
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s s:
s:
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§ o
o
(N O t/5 O ^
P P P o u o
o o o O o o o o o o VO ON ON r- oo o (S fS (S CN (S ( S
d d d d d d d d d d
o o o o o o o o o o Tj- Ti- r j - i n 00 m vo so
es <s >—1 ts ( S ( S
d d d d d d d d d d
o o o o o o o o o o ts t-- (S ON t o 00 iri
fS (N <N (S <N d d d d d d d d d d
o o o o o o o o o o vo r- o 00 vo <s fN ( S I—1 CM r-<
d d d d d d d d d d
u
to •*««» tn C
I to ^
c E cs p
4> E
c o to <0 1» £ Q O I r)
P u
c o OT a u w w E as VI u
c c 0) u 6 E ea 03 V u i: t
£ G Q u u
P P O U
•o — i> o
O. U
0 8 0 7
0 6
0 5 0 4' 0 3 '
0 2-0 1
0
A.indica.
hi. M W
08" 0 7
0 6
0 5 0 4 0 3 -
0 2
0 1
0
0 8
0 71
0 6
0 5i 0 4
0 3 1
0 2
0 1
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C.fistula.
U U I T.indica,
JLLI
M W
M W
0 8
0 7
0 6
0 5
0 4
0 3
0 2
0 1
B Control • Polluted
F.religiosa. 0 8
0 7
0 6
0 5
0 4
0 3
0 2
0 1
0
F.hengalensis.
S - Summer M - Monsoon
ILII S M W
W- Winter
Fig.39. Seasonal alteration in the amount of sodium (mg/gm dry weight) in the wood samples of various tree species.
o o o o
—i—
I
fi o o
I
1
lillliillllliil
_ _ 1
I €
§ O e I
t
1
e
s 1
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•c es >
s k V o ei)
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I c
b. < til
o
s
§ s o •V
o t o
level shows a decline from summer to winter. In A. indica, the amount of sodium
takes deep dip in monsoon and raises in winter.
The Fig. 40 on per cent variation worked out on the basis of sodium content
in the sap wood samples of the different species studied indicates that there has been
a negative variation in the level of sodium in A. indica and F. religiosa under
pollution stress, while in C. fistula and F. bengalemis the negative variation
recorded over the control is only marginal and non-significant. However in T.
indica, there has been a positive variation under pollution stress, although it is
non significant.
The seasonal mean of sodium given in Fig. 40 shows that, A. indica, C.
fistula, F. religiosa and F. bengalensis have experienced significant depletion in
the sodium content in their sap wood samples under pollution stress, while in T.
indica, the sodium level increases to the extent of 35.29% under the same
environmental condition.
CALCIUM IN LEAVES
The data on the contents of calcium (per cent dry weight) in the leaves
collected in various seasons from the species investigated in the present study are
summarized in Table-31. The data reveal that A. indica and T. indica have
accumulated an increased amount of calcium in their foliage under pollution stress,
while in the rest of the species, the level of calcium has fallen significantly compared
to control.
The Fig. 41 indicates the seasonal alteration in calcium balance in the different
species. A glance at the figure clearly shows that in A. indica and T. indica, there
has been an apparent fall in the level of calcium from summer to winter, but not to
the extent of statistical significance. However, in C. fistula, the gradual fall in the
level of calcium has been found to be significant. The trend has been slightly
42
M V > cs
JS JSP '5
t •a e V u 1m
Q.
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"es u
s o E <
ee H
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« E E ss Itl
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I I I — CN r Q Q Q U U U
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(S — m \r\ OS OO O O O d d d
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O O P U O CJ
C/3 C/5 d z
I • ' ^ f^ P P G U CJ u
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s:
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«u o
rj-VO ^ f^ o — d d d P P P U O U
VO tN vO rr o vo fS fo VO rj- VO m o (S 00
rt m CO <o Tf r-> fo
00 VO VO oo o o O o fS O iri 00 iri VO ts 00
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II II II M W
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6 5-
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C.fistula.
JUUI M W
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2.
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T.indica.
JUU M W
F.reUgiosa.
M
B Control • Polluted
W
S - Summer M - Monsoon
F.bengalensis.
M W
W - Wmter
Fig.41. Seasonal variation in the calcium content ( per cent dry weight) in the leaves of various tree species.
disturbed in Ficus spp. in having the peak in monsoon under the pollution stress.
However, the seasonal variation occurring in F. religiosa has been found non-
significant as in the case of A. indica and T. indica.
The Fig. 41, on per cent variation worked out on the basis of calcium content
in the foliage of various investigated species, shows that there has been a significant
fall in the level of calcium in the leaves of C. fistula and F. bengalensis, while in F.
religiosa it is non-significant. In C. fistula and F. bengalensis the highest
percentage of variation has been recorded in the winter samples, amounting upto
12.74% and 19.02% respectively. However, the positive variation registered over
the control in A. indica and T. indica has also been found statistically non-
significant.
The seasonal mean given in Fig. 44 shows that, C. fistula And Ficus spp. have
experienced significant depletion in the calcium content in their foliage under
pollution stress (9.06%, 5.41% and 9.80%), while in indica and T. indica, there
has been a significant increase in calcium content under the pollution stress.
CALCIUM IN BARK
The calcium content in the newly formed bark, on analysis, has shown that,
it increases to a level of significance under pollution load in all the investigated
species with the single exception ofF. religiosa in which the calcium level falls under
the pollution stress. (Table-32).
The Fig. 42 indicates the seasonal variation in the content of calcium in the
bark tissues of the investigated species. It becomes clear that in all the investigated
species there has been a gradual increase in the amount of calcium from summer to
winter under pollution stress. However, the interseasonal variation recorded in the
present study happens to be non-significant in the case of A. indica and C. fistula.
The Fig. 44 on per cent variation denotes that in all the investigated species
43
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o 00 ro o o VO VO o iri (S 1—1 00 o\ r-00 VO rr </-) r—' fO ts ts cs (N m SO
o (N o o o o O o o O cs o\ O tN (N
ON 00 iri Ov vo tn 00 ON 00 ri fs tsi ri cn vo vd TT
cs o o o o o o O vri in o <N 00 vo 00 fS iri iri (S c<i cs <si cs vb vd
o o o O o o o O o o (s vo i ON vo 00 o
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A.indica. C,fistula.
M W M W
T.indica.
Jl M W
F.religiosa.
S M
Control Polluted
W 0
S - Summer M - Monsoon
F.bengalensis.
M W W - Wmter
Fig.129. Seasonal variation in the calcium content ( per cent dry weight ) in the wood samples of various tree species.
there has been a positive variation of calcium observed in their bark samples with the
single exception of F. religiosa in v hich a significant fall on the negative side has
been recorded in their v^inter samples. The variation has been found to be the highest
in summer in case of/4, indica and F. bengalensis, amounting upto 4.96% and 9.74%,
respectively. In T. indica on the other hand, the peak percentage of calcium
(18.38%) has been found to occur in monsoon. However, in C. fistula \ht variation
in the level of calcium recorded over the control in all the seasons has been found
to be statistically significant.
The Fig. 44 on seasonal mean of calcium reveals that in A. indica, C. fistula,
T. indica and F. bengalensis there has been an increased percentage of calcium
accumulation to the extent of significance under pollution stress (4.09%, 7.28%,
12.14% and 5.68% respectively), while in F. religiosa a significant depletion of
3.33% has been recorded.
CALCIUM IN WOOD
The data (Table-33) on the effect of coal-smoke pollution on the calcium
content of the sap wood samples of the selected species in the different seasons imply
that in C. fistula and in the summer and monsoon samples of F. bengalensis there
has been an increase in the amount of calcium under pollution stress, while in the rest
of the species investigated a significant fall in the level of calcium has been recorded.
The Fig. 43 on seasonal deviation in the balance of calcium in the sap wood
sampels of the various investigated species points out that there has been a gradual
increase in the level of calcium from summer to winter in all the species investigated
except in T. indica in which the monsoon samples recorded the peak. However, the
seasonal variation in respect to their calcium content in indica and F. religiosa has
been found to be statistically non-significant.
The per cent variation worked out on the basis of the calcium content of the
44
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m O U
TS O O ^ V
JS •Sf 'S
h. •o E V u u a
E s
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o rs vo o o o o d d I I I
— r j O Q Q O U U
fS — (N \ri O oo O O O d d d
Q Q Q O U U
rr o en c/2
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d d d d d 1—1 f—^ d d
0 0 i n r o v o o o o w-i c - t - v o rri o •ri VO i n m ( S 0 0 0 0
d d d d d d d d
u
s :
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d B « u <o e eo tf)
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A.indica. 1 6 -
C,fistula.
1 4-1 2
1 0-
0 8-
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04-0 2 0 ILJUI
S M W
1 4-) 2 1 0 -
0 8-
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04-02J 0
M W
T.indica. 1 6-
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S M E Control • Polluted
W
1 6-
1 4 1 2
1 0 0 8 0 6 -
04 0 2
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S - Summer M - Monsoon
F.bengalensis.
M W W - Wmter
Fig.43. Seasonal variation in the calcium content ( per cent dry weight ) in the wood samples of various tree species.
2 g J L
VO - i -
o o
o o o i l i l lM
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sap wood samples of the selected species is given in Fig. 44. It manifests that in A.
indica, T. indica and F. religiosa there has been a negative variation in the level of
calcium under pollution stress, and the variation has been found to the extent of
significance only in T. indica with the maximum occurring in monsoon (27.25%).
The positive variation of calcium recorded in C. fistula and F. bengalensis in the
different seasons has been found statistically significant.
The seasonal mean of calcium in the wood samples given in Fig. 44 indicates
that in A. indica, T. indica, F. religiosa and F. bengalensis there has been a
significant loss of calcium in sap wood upto the extent of 19.42%, 21.07%, 5.03%
and 7.39% respectively. While in C. fistula the calcium percentage increased to a
level of significance (23.24%) under polluted condition.
ASCORBIC ACID IN LEAVES
The data, on the effect of coal-smoke pollution on the ascorbic acid contents
in the leaves of the various investigated species show that, the level of ascorbic acid
falls considerably in all the investigated species under pollution stress (Table-34).
The Fig. 45 indicates the seasonal trend in the level of ascorbic acid in the
different investigated species. It becomes clear that in all the investigated species,
there has been a gradual increase in the level of ascorbic acid with the growing age
of the leaves, irrespective of the level of pollution, although the interseasonal
variation has been found to be non-significant in all the species studied.
The per cent variation over the control given in Fig. 46 shows that, the per cent
variation of ascorbic acid contents is negative under pollution stress in all the
investigated species in all seasons. In T. indica, F. religiosa and F. bengalensis the
per cent variation widely varies in summer i.e. in young foliage (38.65%, 45.07%
and 47.79%) and the variation narrows down in older leaves in winter, while in
C. fistula, a more or less to the same extent (21%) of ascorbic acid content remains
45
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Fig.45. Seasonal variation in the amount of ascorbic acid (mg/100 gm fresh weight) in the leaves of different tree species.
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fall in ascorbic acid recorded over the control in A. indica has been very extensive
in summer (31.27%) and narrowed down in winter (21.36%).
The seasonal mean of per cent variation occurred is represented in Fig. 46
and it reveals that, the investigated species have experienced considerable loss of
ascorbic acid in their foliage under pollution stress. The degree of loss in
decreasing order falls as F. religiosa (42.54%)< F. hengalensis (41.63%) < T.
indica (37.88%) < A. indica (26.89%) < C. fistula (21.12%).
PROLINE IN LEAVES
The proline content in leaves has been analysed and the data obtained are given
in Table-35. The analysis shows that the level of free proline increases significantly
in the foliage of all the investigated species under the influence of pollution stress
with the exception of A. indica in which there has been a significant loss in the
production of free proline under the same level of coal-smoke pollution.
The seasonal trend in the level of proline in the different species studied
depicted in Fig. 47 shows that, the level of free proline in leaves reaches its summit
in the winter foliage of all the species after a sharp fall in the monsoon both in control
as well as in those facing the coal-smoke pollution. But, the interseasonal
variation in free proline content of leaves in T. indica, C. fistula and F.
bengalensis has been found to be statistically non-significant.
The per cent variation calculated over the control, on the basis of the amount
of free proline in the foliage of the different species under study is shown in Fig.
48. The figure elucidates that the variation has been positive in the production of
free proline under pollution stress except in /W/ca in which the variation falls
on the negative side. The variation widens in the young foliage i.e. in summer
(67.80%) in case of C. fistula and in the monsoon foliage (49.43%,28.50%) in case
46
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Fig.47. Seasonal variation in the amount of proline (/imoles/g fresh tissue) in the foliage of different tree species.
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of T. indica and F. hengalensis, while in F. reUgiosa maximum variation occurs in
the older leaves (30.92%). However in A. indica the negative variation becomes the
highest in monsoon foliage (38.32%).
The seasonal mean given in Fig. 35 reveals that, there has been a considerable
gain in the amount of free proline in the foliage of C. fistula, T. indica mAF. religiosa
and F. hengalensis (52.70%, 40.18% ,23.31% and 12.96%) under pollution stress,
while 'mA. indica, there has been a significant fall in the level of free proline as a result
of coal-smoke pollution.
47
^ISCUSSIOfhC
D I S C U S S I O N
SULPHUR
Sulphur is an essential element as it forms a constituent ofnumber of amino acids
(Devlin and Witham, 1986). The inorganic sulphur taken up by theplants enter the system
in organic form. The excess ofsulphur is usually accumulated in sulphate form (Thomas
etaL, 1950;Jones, 1962; Decormis, 1969;Fallere/fl/., 1970;HaIlgren, 1978;Thoiron
eta!., ]981;Cram, 1983;Pandey, 1983; Leggee/^?/., 1988) and get eliminated in due
course of time. When they are stored in leaves, they get eliminated when leaves fall and
similarly those stored in bark get eliminated when the bark peels off year after year
(Garsed, 1984; Rennenberg, 1984; Lone, 1993).
The plants in general, when they are grown in sulphur rich atmosphere absorb
excess amount of the element and accumulate in different parts, what has not been
incorporated in the organic system. There are a number of reports regarding the
response of different plants to SO^ exposure under control conditions (Bleasdale,
1952;Tingeye/a/., 1971; Khan and Malhotra, 1977;Lauenroth era/., 1979;Pandey
and Rao, 1978; Garsed o/., 1981; Elkiey and Ormrod, 1981; Koziol and Whatley,
1984; Jagere/o/., 1985; Farooq e/a/., 1988; Beg and Farooq, 1988; Van der Stegen
and Mj'ttenaere, 1991; Wookey and Ineson, 1991; Fuehrer e/of/., 1993) as well as to
SO^ enriched ambient atmosphere under field conditions (Jones et a!., 1974: Wood
and Bonimamu 1975; Ziegler, 1975; Legge e/o/., 1977; 1978; 1980; 1981; 1988:
Pandey. 1983; Trilica o/., 1985: Amundson f /a / . , 1986; 1990; Zech e/«/., 1985,
1990/91: Rao and Dubey 1990 a. b).
When the plants are exposed to SO^. the gas enters the leaf system along with
the air through the stomata and get dissolved in water and affect the working of the
organic system of the cells, disturbing the pH of the medium as well as interfering
the metabolic activities ofthe cells (Malhotra and Sarkar, 1979;Garsede/flr/., 1981; Yu
and Wang, 1981; Trilicae/a/., 1985; Rao and Dubey, 1990a).
The present study to evaluate the sulphur content of the leaves, yielded data
which has brought to light that the selected species have absorbed excess sulphur in
the form of SO^ and accumulated in leaves as well as in other parts of the plant namely
bark and wood. The amount of sulphur accumulated depends on the species as well
as the organs in which they are stored. In general the excess sulphur has been
accumulated as a storage product in leaves and less so in bark and wood. Similar
observations have been recorded by a number of earlier workers in various broad
leaved as well as in conifers (Prasad and Rao, 1982; Pandey, 1983; Alekseev and Rak,
1985; Murray, 1985;Zeche/a/., 1985;Iqbal, 1988; Singh and Rao, 1988; Murray and
Wilson, 1990, 1991; Rao and Dubey, 1990 a, b; Sharma and Prakash, 1991; Van der
stegan and Myttenaere, 1991; Zech e^a/., 1990/91; Lone and Ghouse, 1993;Kupka,
1993; Sah and Meiwes, 1993; Wulfe^ a/., 1993).
The results obtained in the present study on seasonal variation in the sulphur
content of leaves, bark and wood have shown that the seasonal conditions prevailing
in and around the site of study has a profound effect on the sulphur accumulation
in general (Fig. 8-10). In summer, when the growth starts after winter break, the
sulphur content in young tender leaves is found to be the minimum, while in monsoon,
when the leaves become fully grown and mature, the sulphur accumulation becomes
the highest and later recedes in winter. This trend of change in sulphur content
appears to be related to the seasonal conditions as well as to the habit of the species.
In the evergreen species it appears that the seasonal condition and the habit of the
species play a higher role than any other factor. In the heavy and dense atmosphere
of monsoon season, with high humidity, these species catch more SO^ and accumulate
high quantities of sulphur. The same species after monsoon lose some of the
accumulated sulphur and show lesser amount in the older leaves of winter. This state
A9
of affair occurs in the control set as well as in the one which faces pollution, indicating
the phenomenon is more related to the habit of the species than the pollution level
of the atmosphere. In an attempt to study the coal-smoke pollution in the same
experimental site, on some tropical trees Lone (1993) has found that the maximum
concentration of sulphur occurring in monsoon season in Mangifera indica, Psidium
guajava, Syzygium cumini And Eucalyptus citriodora, all having evergreen habit, and
attributed to the active period of growth of these species. The works of Huttunen
etal., (1985) on Scot pine andPahkala (1986) on Hypogymnia spp. have also shown
that these two temperate evergreen species also behave like the tropical evergreen
ones worked out in this laboratory. Contrary to the above, the deciduous Cassia
fistula shows a continuous increase in the amount of inorganic sulphur in its foliage
with the growing age of the foliage culminating in late winter leaves. Lone (1993) has
also reported that in case of the deciduous Dalhergia sissoo and Tectona grandis,
a similar state of affairs in sulphur accumulation as it is noted in the present study
in the case of the deciduous Cassia fistula.
It appears that the phenomenon of sulphur acumulation is more related to age
factor of the leaves than that of seasonal condition of the ambient atmosphere in the
control as well as in the population facing the pollution problem. Lone (1993) has
interpreted the behaviour to be an adaptive measure to avoid excess accumulation of
sulphur in the species worked out by him namely Dalbergia sissoo and Tectona
grandis which drop off their leaves at the time of winter break under the local climatic
conditions.
The results obtained in the present study on sulphur accumulation in newly
formed bark as well as the sap wood of the selected species indicate that the amount
of sulphur content is far less in the bark and in the wood compared to leaves in all
seasons. In the sulphur enriched atmosphere the investigated species showed a
positive accumulation over the control in all seasons, but the degree of accumulation
50
depending on the species. The deciduous species {C. fistula) showed a high amount
of sulphur under coal-smoke pollution in the wood. While in the rest (All evergreen)
the sulphur content varied in wood to a lesser extent compared to control. The
sulphur content in bark, on the other hand, does not show any correlation with the
habit of the species. The evergreen broad leaved Ficus bengalensis shows the highest «
variation in its sulphur content in bark over the control and this is closely followed
by the deciduous broad leaved C. fistula and the evergreen Ficus religiosa. The
minimum amount of sulphur has been found in small leaved Tamarindus indica. This
may be due to the smaller leaf size of the species rather than the habit of the trees.
Among the evergreen species only Azadirachta indica shows significant
amount of sulphur accumulation in wood as well as in bark under coal-smoke
pollution, while the rest have no significant variation in sulpur content of wood,
although all of them have significantly high content of sulphur in the newly formed
bark tissues with the exception of T. indica.
51
C H L O R O P H Y L L PIGMENTS
Chlorophylls, the green pigments of plants are the most important pigments
responsible for the conversion of light energy into chemical energy and are thus active
in the process of photosynthesis. Chlorophyll molecule has a cyclic tetrapyrrolic
structure (Porphyrin), with an isocyclic ring containing a magnesium atom at its
centre and a phytol chain attached to it. Chloroplast of higher plants alv^ays contain
two types of chlorophyll. One is invariably chlorophyll a, and the other is
chlorophyll b, which has an aldehyde group instead of a methyl group attached to
ring II. Most higher plants contain about twice as much chlorophyll a as chlorophyll
b. Carotenoids, on the other hand are also found in varying amounts in nearly all
higher plants and are believed to be vital for two important functions: (a), they
protect against the photo-oxidation of chlorophyll and (b), they absorb and transfer
light energy to chlorophyll a (Devlin and Witham, 1986).
The pigment analysis undertaken in the present study reveal that the ratio of
chlorophyll a and chlorophyll b under normal conditions range from 1:0.65 to 1:0.76
in the species investigated (Table-36). This goes against the common belief that
chlorophyll a is twice the number of chlorophyll b as mentioned by Lehninger et al,
(1992). The pigment contents when analyzed, has revealed that all the investigated
species undergo considerable loss in their coloured pigments under coal-smoke
pollution with the exception of F. bengalensis in which the pigment strategy has
proved to be highly productive under coal-smoke pollution. The loss in chlorophyll
pigments under sulphur dioxide pollution has been a common observation by almost
all workers in the past both under controlled as well as under field conditions.
"(Wellburne/a/., 1972;Malhotra, 1977;Pandey andRao, 1978; Prasad and Rao, 1982;
Devi and Patel, 1983; Pandey, 1983; Irving and Miller, 1984; Malhotra and Khan,
1984; Sharma and Rao, 1985; Singh e/a/., 1985; Yunusetal.,\9B5, Kumar and Yadav,
1986; Nandi etai, 1986; Singh and Rao, 1986; Agrawal etal, 1987; Chand and Kumar,
1987; Gupta and Ghouse, 1987b; Beg and Farooq, 1988; Farooq e/a/., 1988; Singh
and Rao, 1988; Vijayan and Bedi, 1988; Prakash a/.. 1989a; Singh, 1989; Saquib
and Ahmad, 1991; Sharma and Prakash, 1991; Ghouse et al., 1993;
Ramasubramanian et al, 1993; Sabu etal, 1993; Sarinen and Liski, 1993). The
reduction in chlorophyll contents by sulphur dioxide, hydrogen fluoride and
hydrogen chloride is a generally accepted fact, as these compounds create low pH
of the medium which eventually brings about decolourization and degradation of
pigments (Arndt, 1971;Puckete/a/., 1973; Devi and Patel, 1983). But the observations
recorded in the present study on Ficus bengalensis appears to be unique to this
species, as it experiences significant gain in the amount of chlorophyll pigments and
carotenoids under coal-smoke pollution, instead of undergoing any loss. A similar
situation of increased chlorophyll contents has been come across by Fulford and
Murray (1990) in two species of Eucalyptus under SO^ pollution. Earlier, Murray
and Wilson (1988) have also demonstrated the increase in chlorophyll concentration
in Eucalyptus tereticornis when exposed to concentrations of SO^ and hydrogen
fluoride. Doley (1988) has found increase in chlorophyll concentration in four
species oiPinus under fluoride pollution. Devi and Patel (1983) have also found
increased amount of chlorophyll a, chlorophyll b and total chlorophyll in a number of
species including Ficus bengalensis under the pollution caused by fertilizer complex
of Baroda. The gain in pigment concentration found in F. bengalensis has been
reflected in the higher carbohydrate content under coal-smoke pollution, discussed
elsewhere in detail. Devi and Patel (1983) have also found increased sugar
production in plants in which there has been an increase in chlorophyll concentration
under pollution stress. The correlation coefficient worked out between sulphur and
photosynthetic pigments in the investigated species given in Table-37, has clearly
revealed that the degree of dependence of these pigments over sulphur is quite high
53
in all species. All the species showed a negative correlation with the exception ofF.
bengalensis which depicted positive correlation.
Among the chlorophyll pigments analysed, chlorophyll 'a ' suffered greater
loss in C. fistula and T. indica in all seasons, while the rest of the species the chlorophyll
'b ' undergoes higher losses. The original ratio of chlorophyll 'a ' and chlorophyll 'b'
under normal atmosphere, thus get disturbed and altered. Under pollution stress the
ratio of the chlorophyll 'a ' to chloropyhll 'b' ranges from 1:0.6 to 1:0.77 (Table-36).
The low ratio of chlorophyll 'b' has been found in Azadirachta indica and Ficus
religiosa due to higher loss of chlorophyll 'b' than chlorophyll 'a ' . In Cassia fistula
and Tamarindus indica the ratio of chlorophyll 'b' improves over chlorophyll 'a ' due
to higher loss of the latter. Whereas, in the case of Ficus bengalensis, the position
of chlorophyll 'b ' becomes stronger due to its higher production than chlorophyll
'a ' under coal-smoke pollution.
The ratio of total chlorophyll in the investigated species falls under coal-
smoke pollution with the exception of F. bengalensis in which the ratio increases by
0.3 over the control. The higher loss of chlorophyll 'a ' compared to chlorophyll 'b'
has been interpreted to its high sensitivity to SO^ pollution by earlier workers
(Sunderland, 1967; Malhotra, 1977; Kondoe/a/., 1980; Chaudhary and Sinha, 1982;
Periasamy and Vivekanandan, 1982; Singh and Rao, 1986; Gupta and Ghouse, 1987b,
Khan and Usmani, 1988; Prakash et al, 1989b; Singh, 1989). The greater loss of
chlorophyll 'a ' thr.n 'b' has been interpreted in more than one way (Khan and Usmani,
1988) and it is ascribed to the inactivation of various enzymes associated with the
synthesis and action of chloropyhll 'a ' . Knudson et al, (1977) suggested the
following possibilities for the reduction of chlorophyll a to chloropyhll b ratio due
to the exposure of plants to ozone;
(a) Chlorophyll 'a ' may degrade more rapidly than chlorophyll 'b ' ; and (b)
either the synthesis of chlorophyll 'a ' may be reduced or that of chlorophyll 'b ' may
54
be increased (Godnev and Shabel's Kaya, 1963). Chlorophyll 'b' may be formed from
chlorophyll 'a ' , and chloropyhll 'a ' may undergo rapid degradation as compared to
chlorophyll 'b ' in senescent leaves (Wolf, 1956).
The higher sensitivity of chlorophyll 'a ' to pollution stress hampers the plant
growth considerably as it plays a very important role in the process of photosynthesis
(Malhotra, 1977). Comparatively higher losses of chlorophyll 'b ' is found in the
present study in A. indica and in F. religiose need special mention. Devi and Patel
(1983) has also found higher reduction in chlorophyll 'b' in A. indica as it has been
found in the present study while working on the effect of air pollution emitted by
a fertilizer complex situated in Baroda.
The seasonal relationship in the chlorophyll content of leaves found in the
present study in case of A. indica and T. indica, both happened to be evergreen
species, may be due to the induced early senescence by the pollutants. The higher
loss of chlorophyll pigments in the young leaves of summer as found in Cassiafistula
in the present study may be due to the higher sensitivity of young foliage than the
older ones. What is found in the other species the higher loss of pigments in the
monsoon season appears to be normal as the noxious gases get higher entry into the
leaves in the dense and heavy atmosphere of the season due to the delayed and slow
diffusion and dispersal of these gases in the atmosphere.
55
Table 36. Ratio orPhotosynthetic Pigments.
SPECIES SITE PIGMENTS RATIO
A. mdica P Chi a b 1 0 61 C Chi a b 1 0 70
C. fistula P Chi a b 1 0 71 C Chi a b 1 0 65
T. mdica P Chi a b 1 0 77 c Chi a b 1 0 67
F. rebgiosa p Chi a b 1 0 70 c Chi a b 1 0 76
F. hengalensis p Chi a b 1 0 71 c Chi a b 1 0 67
A. mdica C P Total Chi 1 0 86
C. fistula C P Total Chi 1 0 76
T. mdica C P Total Chi 1 0 73
F religiosa C P Total Chi 1 1 30
F. bengalensis C P Total Chi 1 080
A. mdica p Total Chi Crotenoid 1018 c Total Chi Crotenoid 1 020
Cfistula p Total Chi Crotenoid 1025 c Total Chi Crotenoid 1026
T. mdica p Total Chi Crotenoid 1 0 26 c Total Chi Crotenoid 1 022
F. religiosa p lotalChl Crotenoid 1 023 c Total Chi Crotenoid 1 0 23
F. bengalensis p Total Chi Crotenoid 1 0 22 c Total Chi Crotenoid 1 026
P = Polluted, C = Control, Chi = = Chlorophyll
Table 37. Correlation coefllcient (r) between Sulphur and Photosynthetic Pigments.
SPECIES PIGMENTS SEASONS
Summer Monsoon Winter
A. indica Chi. a Chl.b Carotenoids Total Chi.
-0.8992* -0.9403** -0.3789 -0.7738
-0.9731** -0.9447** -0.9451** -0.9679**
-0.9489** -0.9670** -0.9627** -0,9693**
C. fistula Chi. a Chl.b Carotenoids Total Chi.
-0.7983 -0.8587* -0.8210* -0.8548*
-0.8449* -0.5724 -0.8974* -0.5589
0.9328** -0.7913 -0.6694 -0.9264**
T. indica Chi. a Chl.b Carotenoids Total Chi.
-0.9749** -0.9789** -0.9964** -0.9773**
-0.9904** -0.9788** -0.9533** -0.9913**
-0.9722** -0.8788* -0.9141* -0.9674**
F. religiosa Chi. a Chl.b Carotenoids Total Chi.
-0.7938 -0.9505** -0.9523** -0.3262
-0.4501 -0.9002* -0.9595** -0.9958**
-0.9219** -0.9617** -0.9498** -0.9501**
F. bengaletisis Chi. a Chl.b Carotenoids Total Chi.
0.7273 0.8474* 0.8578* 0.8626
0.8289* 0.1694
-0.2811 0.8785*
0.8737* 0.8036* 0.8579* 0.7118
* Significant at 5% level ** Significant at 1% level Chl.= Chlorophyll
CAROTENOIDS
Carotenoids are lipid compounds that are distributed widely in both animals
and plants and range in colour from yellow to purple. Carotenoids are present in
variable concentrations in nearly all higher plants. They absorb light at wave lengths
other than those absorbed by the chlorophylls and thus act as supplementary light
receptors. The major carotenoids found in plant tissues is the orange-yellow pigment
/3-carotene, which is generally accompanied by varying amounts (0 to 35 per cent)
of a carotene (Mackinney, 1935). The probable roles of carotenoids are (1) They
protect against the photooxidation of chlorophylls and (2) They absorb light and
transfer the energy to chlorophyll a (Devlin and Witham, 1986).
The observations recorded in the present study on the carotenoid contents
show heavy losses of carotenoids in the winter foliage of Azadirachta indica,
Tamarindus indica and Ficus religiosa. In a recent study on some tropical trees
exposed to coal-smoke pollution Lone (1993) has also found a similar behaviour of
foliage in respect to carotenoids. He has also found that the loss of carotenoids
amounts to be higher than the total chlorophyll in old leaves of winter as found in
the present study in case of Azadirachta indica and Tamarindus indica. Nouchi et
al. (1973) and Prasad and Rao (1982) have also noted higher losses of carotenoids
in older leaves than in the young ones. The higher loss of carotenoids in the young
foliage of Cassia fistula found in the present study is going in accordance with the
findings of Lone (1993) on Dalbergia sissoo and Syzygium cumini, indicating the
behaviour of carotenoids to be independent of age factor and their behaviour
depending on other factors like genetic constitution of the concerned species.
The ratio (Table-36) of total chlorophyll to carotenoids in the investigated
species varies from 1:0.22 to 1:0.26 in the control and from 1:0.18 to 1:0.26 in the
polluted site. In Tamarindus indica, the ratio of total chlorophyll and carotenoids occurs
at 1:0.22 in control and the ratio of carotenoids increases to 0.26 in polluted samples; in
F. bengalensis it remains the same while in others its ratio decreases to varying degrees.
In T. indica the tremendous decrease (26%) in cholorophyll contents in the polluted
samples boosted the ratio of carotenoids, although the latter also get decreased due to
pollution by 16%. The gain in total cholorphyll and carotenoids under pollution is almost
the same which maintains the ratio to the same extent in F. bengalensis. The higher loss
of carotenoids (33%) in F. religiosa disturbs its ratio under pollution stress to reduce it
to the minimum in this species. Although the loss of carotenoids amounts to 25.74% in
C. fistula, the ratio is not disturbed (0.01%) in this species, as the loss in total chlorophyll
also amounts to a great extent i.e. 22.51% indicating the carotenoids as well as the
chlorophyll to be highly sensitive in this species to coal-smoke pollution. No work in this
line to study the ratio of carotenoids to chlorophyll under pollution appears to have been
undertaken so far, as there is no report in the literature.
57
CARBOHYDRATE
The carbohydrates are important to the plant in several ways. First, they represent
a means for the storage of the energy, second, they are important constituents of the
supporting tissues that enable the plant to achieve erect habit, third, they provide the
carbon skeletons for the organic compounds that make up the plant (Devlin and Witham,
1986).
Plant grovk'th and development represent a complex series of co-ordinated
events which are ultimately dependent upon the organic reserves accumulated in the
seed (Cotyledons) while, the subsequent growth depends upon the translocation of
carbohydrate in excess of their maintenance needs. The productivity of mature
leaves, therefore represent an asset to the total carbon economy of a plant while
developing leaves, flowers and fruits represent liabilities requiring an input of carbon.
In most cases, the productivity of mature leaves is more than sufficient to meet these
demands of carbon for growth and development. Exposure to gaseous pollutants can
however, alter the balance of a plant's carbon economy so that growth can be retarded
and yield reduced. In the case of chronic exposures, reductions in growth and yield
can occur without accompanying visible symptoms of injury.
In the present study the carbohydrate content in the leaves of the investigated
species has been found to be higher in the older leaves of monsoon and winter seasons
than in the young summer foliage in all the species both in the control as well as in
the polluted samples. In Ficus bengalensis under pollution stress there has been a
significant increase in the carbohydrate content in all seasons. This increase in
carbohydrate content appears to have been directly related to the increase in the
chlorophyll and carotenoid contents of this species under pollution stress.
In case of 71 indica, there is a fall in the level of carbohydrate in leaves, bark
and wood under pollution stress, as there has been a significant reduction in the
amount of chlorophyll and that of carotenoids in this species consequential to coal-
smoke pollution. However in the rest of the species, there has been an increase in
the carbohydrate level under pollution, although there has been a deep fall in the level
of coloured pigments in the leaves in all seasons. There are several reports on SO^
influenced reduction in the carbohydrate contents in different plant species viz.
Ulmus americana (Constantinidou andKozlowski, 1979); Triiicum aestivum (Prasad,
1980); Pinus sirobus (Percy and Riding, 1981); Glycine max and Triiicum aestivum
(Prasad and Rao, 1982); Avena sativa (Chand and Kumar, 1987); Syzygium cumini
(Vijayan and Bedi, 1988); Lycopersicon esculentum and Hordeum vulgare (Prakash
et al, 1989a, b) and Polyalthia longifoUa (Singh, 1989). Hovs^ever the increase in
carbohydrate contents recorded in the foliage of various investigated species in
different seasons at the polluted site in the present study may be due to the lesser
utilization of carbohydrate due to reduced growth activity of the plants as well as
inhibition of other metabolic processes involved in carbohydrate metabolism. Lone
(1993) has also recorded the increase in carbohydrate level in different seasons of
some fruit trees as well as in certain timber trees growing under the stress of coal-
smoke pollution, though there has been significant reductions in total chlorophyll and
carotenoids in these species. The accumulation of carbohydrate in the foliage under
SOj enriched condition may be also due to the inhibition in its translocation from the
leaves (Noyes, 1980), probably because of the phloem loading due to SO^ pollution
(Koziol, 1984).
The data obtained on correlation coefficient between total chlorophyll and
carbohydrate content (Table-38) indicate that the correlation is positive in case of
F. hengalensis and T. indica irrespective of seasonal variation. In case o f ^ . indica
the correlation between chlorophyll and carbohydrate content is negative as these
two aspects run against each other. That is to say that the carbohydrate content in
leaves records an increase against the fall of chlorophyll level. In the rest of the
59
species, there is a positive correlation between carbohydrate content and chlorophyll
level in the young leaves of summer but not in the older leaves of monsoon and winter
seasons.
The correlation coefficient worked out between carotenoid contents and
carbohydrate also shows the same trend as it has been found between chlorophyll
and carbohydrate (Table-18).
The correlation between sulphur content in the leaves and carbohydrate is
summarized in Table-39. In A. indica and F/ci/5 spp., the correlation shows perfectly
positive in all the seasons. However, in C. fistula the level of carbodydrate in summer
foliage is negatively correlated with sulphur (r=-0.85), and the rest of the seasons
records positive correlation. In T. indica, the correlation between sulphur and
carbodydrate is negative in all the seasons, running opposite to each other. The 'r '
value ranges from 0.65 to 0.83 in this species.
The concentration of carbohydrates in bark and wood in the present study
reveals that there has been an increase in the level of carbohydates in the bark samples
of A. indica, C. fistula dinAF. under pollution stress, but the wood samples
of these spp. records considerable loss in all seasons. However, in T. indica and F.
bengalensis considerable loss in carbohydrate occurs in bark tissues under pollution
stress. In the wood samples of T. indica there has been a significant loss in
carbohydrate while in the wood ofF. bengalensis there has been a considerable gain
in carbohydrate level. The increase in carbohydrate recorded in the bark sample of
A. indica, C. fistula and F. religiosa under pollution stress in the present study is in
confirmity with the earlier report of Lone (1993) on Mangifera indica, Psidium
guajava and Tectona grandis. The loss of storage products like carbohydrate in
wood and bark under pollution stress is not uncommon in literature. Patel and Devi
(1986) observed reduced levels of starch in the bark and wood samples of Syzygium
cumini and Mangifera indica populations growing in the polluted atmosphere around
60
a fertilizer complex. The loss of carbohydrate observed in the bark samples of T.
indica and F. bengalemis and in the wood samples of all the species except in F.
bengalensis in the present study is in agreement with the above report. Similarly the
significant increase in carbohydrate content in the wood samples recorded in F.
bengalensis in all the seasons in the present study is also in agreement with earlier
report of Lone (1993) in some tropical trees {Syzygium cumini. Eucalyptus
citriodora and Tectona grandis).
In the present study, the maximum increase of carbohydrate under polluted
condition occurs in the monsoon samples of A. indica (32.07%) and the least records
in the monsoon samples of C. fistula and F. religiosa (9%). Likewise, the decrease
in carbohydrate touches the maximum in the winter samples of T. indica (25.99%)
and the minimum in the monsoon samples of T. indica and F. bengalensis (6%).
In the present study, the maximum concentration of carbohydrate in the
polluted as well as in normal atmosphere has been recorded in wood compared to leaf
and bark. The loss in carbohydrate experiences the maximum in the monsoon wood
samples of T. indica (35.03%) in the polluted condition, and the least in its summer
samples (6.08%). Similarly, the increased accumulation of carbohydrate under
pollution stress records the highest in the late winter samples of bengalensis
(10.77%) and the least in its monsoon samples (3.99%).
The correlation worked out on sulphur versus carbohydrate content in the
bark and wood shows that, the correlation is positive in the bark samples oiA. indica,
C. fistula and F. religiosa, in all the seasons except in the summer samples of C. fistula
in which there occurs a perfect negative correlation (r=-0.93). In the rest of the
species, bark records negative correlation with sulphur. In wood, all the investigated
species exhibit negative correlation against sulphur irrespective of seasons, with the
exception ofF. bengalensis in which there occurs a non significant positive correlation.
The deciduous C. fistula is the only species to show significant negative correlation
of carbohydrate with sulphur in wood (r=-0.82 to -0.90). The rest of the species
records non-significant correlation (Table-39).
61
Table 38. Correlation coefllcient (r) between Chlorophyll and Carbohydrate.
SPECIES SEASONS
Summer Monsoon Winter
LEAF
A. indica -0.7787 -0.8065* -0.8454* Cfistula 0.8583* -0.5762 -0.8777* T. indica 0.9022* 0.7491 0.6597 F. religiosa 0.9434** -0.9336** -0.9084* F. bengalemis 0.8275* 0.9096* 0.7897
BARK
A. indica -0.9106* -0.9734** -0.9849** C. fistula 0.8389* -0.5971 -0.9496** T. indica 0.9797** 0.9343** 0.9842** F. religiosa -0.9624** -0.8607* -0.9561** F. bengalemis -0.8700* -0.8195* -0.7556
WOOD
A. indica 0.9348** 0.9366** 0.9620** C. fistula 0.8857* 0.7397 0.9095* T. indica 0.9449** 0.9952** 0.9884** F. religiosa 0.8629* 0.9347** 0.9533** F. betigalensis 0.9090* 0.8313* 0.7479
* Significant at 5% level ** Significant at 1% level
Table 39. Correlation coefllcient (r) between Sulphur and Carbohydrate.
SPECIES
Summer
SEASONS
Monsoon Winter
LEAF
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.728] -0.8567* -0.8386* 0.9569** 0.8815*
0.8441* 0.5025
-0.7113 0.9347** 0.9646**
0.8041 0.8762*
-0.6594 0.9005* 0.9759**
BARK
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.7868 -0.9313** -0.8923* 0.8657*
-0.7269
0.1797 0.4257
-0.8579* 0.8832*
-0.2051
0.6567 0.8588*
-0.8070 0.3493
-0.3831
WOOD
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.6092 -0.9071* -0.0854 -0.4668 0.1398
-0.4458 -0.8846* -0.3926 -0.2780 0 1880
-0.5903 -0.8274* -0.2048 -0.3023 0.1769
* Significant at 5% level ** Significant at 1% level
P R O T E I N
The proteins are the most vital components of the living systems and their
most significant influence resides in the fact that many are functionally active as
enzymes which are vital for the rapid rates of biochemical reactions. Proteins also
act as natural hydrogen ion buffers and structural components of the cells. The
literature pertaining to the impact of SO^ on the protein contents of the plants is well
documented. In a recent communication Roa and Dubey (1990 a, b) reported
significantly reduced protein contents in the foliage of several plant species viz.
Azadirachia indica, Calotropisprocera. Cassia siamea, Dalbergia sissoo, Ipoemea
fistulosa, Mangifera indica, Syzygium cumini and Zizyphus mauritiana growing in
field conditions around an industrial area. In an earlier detailed investigation on
certain tropical tree species. Beg and Farooq (1988) observed that SO^ concentrations
which produced no visible injury, decrease the protein contents in Ficus rumphi,
Holoptelea integrifolia, Mangifera indica, Psidium guajava and Syzygium cumini.
However, no change was observed in the Pithecolobium dulce while Ficus bengalensis
shows elevated protein contents.
The present investigation on five tropical trees, the protein level in leaves
increased in four out of the five species. The only species in which a decrease in
protein level takes place, is Tamarindus indica. There are several reports in the past
that recorded reduction in protein content in plants under the stress of SO^ pollution
(Fischer, 1971; Godzik and Linskens, 1974; Mudd, 1975; Constantinidou and
Kozlowski, 1979;MalhotraandSarkar, 1979; Grill a/. 1980; Prasad, 1980; Robe and
Kreeb; 1980, Percy and Riding, 1981; Agrawal, 1982; Singh e/a/. 1985;Krishnamurthy
etal. 1986; Vijayan and Bedi, 1988; Rao and Dubey, 1990 b). The reports showing the
increase in protein content under pollution stress is also not uncommon in literature
(Godzik and Linskens, 1974; Prasad and Rao, 1982; Saxe, 1983; Murray, 1984; Rao
andDubey, 1990a; Lone, 1993). There is a high concentration ofprotein in the leaves
of Azadirachta indica to the extent of57.86% in the young leaves of summer season. In
Cassia fistula too similar trend of high concentration in the young leaves and a reduced
level of the same in the older leaves has been recorded in the present study. This
increased concentration reflects the enhanced synthesis of protein in the young leaves of
these two species which may act as combating mechanism against pollution hazard
(Crakerand Starbuck, 1972; Jager er a/., 1985).
In Ficus bengalemis on the other hand, it has been found that an initial fall in the
protein content in the young summer foliage followed by a sharp increase (94.07%) in
monsoon. In F. reJigiosa the highest amount of protein content has been observed in
winter foliage (47.08%). The fall in the concentration of protein in young leaves under
pollution stress may be due to the changes in amino acid concentration in SO^ treated
plants, leading to the protein reduction (Godzik and Linskens, 1974) due to the
inactivationofenzymes responsible for the protein synthesis (Cecil and Wake, 1962).
Malhotra and Khan (1984) are of the view that the decrease in the protein contents could
be attributed to the break down of existing proteins and to the reduced de novo synthesis.
The reduction in protein contents of SO^ treated plants might also result from decreased
photosynthetic activity (Sij and Swanson, 1974; Constantinidou and Kozlowski, 1979).
The correlation coefficient worked out on leaf sulphur concentration and protein
content shows that the correlation becomes positive in the foliage of all the species in
different seasons except in T. indica and in the summer samples oiF. bengalensis in
which correlation records negative. However, the correlation is not significant in T.
/>ji//ca because the ' r 'value falls in the range of-0.38 to-0.45. Similarly, the positive
correlation recorded in different seasons in the rest of the species also shows non-
significance with sulphur in all seasons except in the winter samples of the foliage ofF.
bengalensis which records a significant correlation (Table-40).
The observations on the protein content of the newly formed bark and wood
63
samples in the present study show that the protein concentration is significantly high
in the bark and wood samples of indica in the SO^ enriched atmosphere, while the
bark and wood samples of C. fistula and F. bengalensis have recorded considerable
loss in their protein content under pollution stress. However, in F. religiosa bark
shows considerable loss in protein content, but an opposite condition prevails in the
case of wood. In A. indica and T.indica both bark and wood have accumulated
appreciable amount of protein in varying degrees under the stress of coal-smoke
poJJution (Table 17-18).
The correlation of protein worked out against sulphur concentration in wood
and bark shows that the ' r ' value is negative in C. fistula and F. bengalensis, while
in F. religiosa, the ' r ' value in the bark is negative, but it becomes positive in the wood
samples. However, in the rest of the species the ' r ' value shows positive in all
seasons. In bark the significant correlation occurs in the monsoon samples of T.
indica and in the monsoon samples of F. bengalensis, while in wood the significant
correlation found only in the monsoon samples of C. fistula (Table-40).
64
Table 40. Correlation coeflicient(r) between Sulphur and Protein.
SPECIES
Summer
SEASONS
Monsoon Winter
LEAF
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.8014 0.7082
-0.3913 0.7795
-0.5479
0.7602 0.2941
-0.4548 0.5518 0.7558
0.5159 0.5699
-0.3846 0.7903 0.8860*
BARK
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.4979 -0.4520 0.9507**
-0.5827 -0.5249
0.3929 -0.4556 0.8352*
-0.3861 -0.8764*
0.1446 -0.7294 0.7758
-0.4560 -0.4560
WOOD
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.4984 -0.5493 0.2794 0.3112
-0.0865
0.5326 -0.8321* 0.2115 0.6242 -0.1440
0.5739 -0.7044 0.4332 0.1081
-0.1008
* Significant at 5% level ** Significant at 1% level
NITROGEN
Nitrogen is one of the essential nutrients for the plants and perhaps nitrogen' s most
recoginsed role in plants is its presence in the structure of protein molecules. In addition,
nitrogen is found in such important molecules as purines, pyrimidines, porphyrins and
coenzymes. Purines and pyrimidines are found in the nucleic acids, RNA and DNA
essential for protein synthesis and transfer of genetic material (Devlin and Witham, 1986).
The porphyrin structure is found in such metabolically important compounds such as the
chlorophyll pigments and cytochromes, essential in photosynthesis and respiration.
Coenzymes are essential to the function of many enzymes. In addition to its absorption
generally in the form of NOj", nitrogen is also taken up as N H / . Once inside the plant,
nitrogen is reduced and incorporated into diverse organic compounds (Beevers and
Hageman, 1969). Its deficiency may result in stunted growth and yellowing of leaves on
account of loss of chlorophyll.
It is an established fact that the foliar level of different inorganic elements are
affected by (i) uptake from the soil and atmosphere (2) translocation to and from
other tissues within plant (3) loss by leaching or volatalization (Van den Driessche,
1974; Miller, 1984; Johnson et ai, 1985). Further, uptake, accumulation and
distribution of various mineral nutrients in the plant body are influenced by several
factors such as climate, season, time of the day, age of the tree and foliage, type of
the tree species and soil conditions (Bazilevic and Rodin, 1964; Ovington; 1908,
Evans, 1979).
The nutritional effect of NO^ on the plants in nitrogen deficient conditions has
been reported by several workers in the past (Troiano and Leone, 1977; Troiano,
1978; Yoneyama et al., 1980; Srivastava and Ormrod, 1984; Okano and Totsuka,
1986; Rowland et al., 1987; Wingsle et ai, 1987; Sabaratnam et al., 1988;
Ramasubramanian ^/o/., 1993). In the present study (Table-19) the nitrogen content
increases in the foliage of all the investigated species except in Tamarindus indica in
which the nitrogen content falls significantly in all the seasons under pollution stress. The
increase in nitrogen contents in plants has also been reported after fumigation with NH^
(Draaijers et al, 1989; Bobbink et al., 1990; Dueck and Elderson, 1992) or after
exposing with acid mist containing major sulphur or nitrogen pollutants (Kimball et al.,
1988; Jacobson et al., 1989). It is an established fact that plant leaves can absorb
different kinds of gaseous air pollutants, including NO^ through their stomata (Spedding,
1969; Rich e/a/., 1970; Porter e/a/., 1972; Okano e/a/., 1989; Pearson e/or/., 1993).
Further, it is also possible that plants may absorb and fix NO^ from the atmosphere and
thus increase their nitrogen contents. Since NO^ is also one of the by products of coal-
burning, its presence in the ambient atmosphere at the test site is unavoidable. The
retarded growth activity of the plants in the polluted atmosphere coupled with lesser
metabolic activities might have left nitrogen in the plant body ut\utilized and thus get
accumulated. However, the lower concentration of NO^ absorbed by the plants can be
easily metabolized and detoxified if relevant enzymes are active (Rogers et al, 1979;
Yoneyama and Sasakawa, 1979; Rowland e/a/., 1987).
The observations recorded in the present study on the seasonal variation of
nitrogen accumulation shows that, in F. hengalensis and F. religiosa there has been
an increased percentage of accumulation of nitrogen in the winter foliage, while the
same condition has been recorded in monsoon in A. indica and in summer in C. fistula.
The increased per cent of nitrogen recorded in the winter samples of the Ficus spp.
is in agreement with the earlier report of Lone (1993) in some fruit trees from the
same site. He interpreted that , as an adaptation developed by the plants so as to
translocate most of it towards the newly emerging leaves which may need higher
nitrogen supply during stressed condition. Sacher (1957) and Das (1968) have also
reported that minerals are transferred to green or living parts when the leaves began
to dry. However, the increased percentage of nitrogen accumulation recorded in other
66
seasons in the present study in two species viz. A indica and C. fistula may be due
to the greater entry and absorption of NO^ from the atmosphere due to the active
growth of leaves in the respective seasons. Several other workers in the past
(Malkonen, 1974; Evans, 1979; Garsede/a/., 1981) have also observed an increase
in nitrogen concentration in young leaves which decreased with the increasing age of
the leaves.
The decrease in nitrogen content in plants under pollution stress is not also
uncommon. In the present study the significant loss in the nitrogen percentage
recorded in Tamarindus indica in all seasons under pollution stress goes in agreement
with the earlier reports in some conifers (Malcom and Garforth, 1977; Garsed et al,
1981) and in some annual crops (Pandey and Rao, 1978; De santo etal., 1979; Mishra,
1980; Elkiey and Ormrod, 1981; Agrawal, 1982; Sharma and Rao, 1985; Prakash et
al, 1989a) in some broad leaved trees like Diospyros melanoxylon, Lagerstroemia
parviflora and Zizyphus nummularia under field conditions around a coal fired power
plant (Pandey, 1983). Zech etal. (1990/91) have also observed decreased values for
'N ' contents in declining populations growing in an SO^ polluted area
of Ne-bavarian mountain.
The correlation coefficient worked out between leaf sulphur concentration
and nitrogen content given in Table-41 shows a positive relationship between
sulphur and nitrogen concentrations in the leaves of all the investigated species
except Tamarindus indica in which both aspects runs opposite because of the
reduced nitrogen content in the foliage under pollution stress. Among them, summer
samples of T. indica and Ficus spp. as well as the monsoon samples of C. fistula and
F. religiosa and the winter samples of A. indica records non significant correlation
with sulphur.
The observations recorded on the nitrogen contents in bark and wood samples
summarized in Table 20-21, show that in^^. indica and T. indica there has been an
67
seasons in the present study in two species viz. A indica and C. fistula may be due
to the greater entry and absorption of NO^ from the atmosphere due to the active
growth of leaves in the respective seasons. Several other workers in the past
(Malkonen, 1974; Evans, 1979; Garsed etal, 1981) have also observed an increase
in nitrogen concentration in young leaves which decreased with the increasing age of
the leaves.
The decrease in nitrogen content in plants under pollution stress is not also
uncommon. In the present study the significant loss in the nitrogen percentage
recorded in Tamarindus indica in all seasons under pollution stress goes in agreement
with the earlier reports in some conifers (Malcom and Garforth, 1977; Garsed et al,
1981) and in some annual crops (Pandey and Rao, 1978; De santo et al., 1979; Mishra,
1980; Elkiey and Ormrod, 1981; Agrawal, 1982; Sharma and Rao, 1985; Prakash e/
al., 1989a) in some broad leaved trees Wke Diospyros melanoxylon, Lagerstroemia
parviflora and Zizyphus nummularia under field conditions around a coal fired power
plant (Pandey, 1983). Zech etal. (1990/91) have also observed decreased values for
'N ' contents in decVmingFagussylvatica populations growing in an SO^ polluted area
of Ne-bavarian mountain.
The correlation coefficient worked out between leaf sulphur concentration
and nitrogen content given in Table-41 shows a positive relationship between
sulphur and nitrogen concentrations in the leaves of all the investigated species
except Tamarindtts indica in which both aspects runs opposite because of the
reduced nitrogen content in the foliage under pollution stress. Among them, summer
samples of T. indica and Ficus spp. as well as the monsoon samples of C. fistula and
F. religiosa and the winter samples of A. indica records non significant correlation
with sulphur.
The observations recorded on the nitrogen contents in bark and wood samples
summarized in Table 20-21, show that in^. indica and T. indica there has been an
67
increase in the amount of nitrogen in the bark and wood samples in all the seasons
under pollutiion stress In 7. mc//ca the concentration of nitrogen in the bark samples
of control and polluted, as well as its per cent variation in all the seasons record higher
than the wood. However, in C. fistula and F. bengalensis there has been a fall in the
level of nitrogen in the polluted bark and wood samples of all seasons. While in F.
religiosa bark showed considerable loss but an opposite condition prevails in wood
in all seasons under SO^ enriched condition.
The correlation worked out between concentration of sulphur in bark and
wood against their nitrogen concentration shows that the correlation is positive in
the bark and wood samples of A. indica and T. /W/ca under pollution stress. The 'r '
value in the bark sample of J. indica is quite high and it ranges from 0.90 to 0. 98.
In A. indica 'r ' value becomes non-significant in the bark. Similarly, in the wood
samples both^. indica and T. /W/ca record poor correlation. The negative correlation
recorded in the bark and wood samples of C. fistula and F. bengalensis is non-
significant, but in C. fistula the summer and monsoon samples of the bark records
perfect negative correlation (r = - 0.92) with sulphur. Likewise, the nitrogen content
of wood samples of C. fistula also has shown significant correlation in its monsoon
samples with sulphur. However, in F. religiosa, the bark has recorded significant
negative correlation in summer and monsoon but an opposite condition prevails in
wood. (Table-41).
6 8
Table 41. Correlation coefTlcient (r) between Sulphur and Nitrogen.
SPECIES
Summer
SEASONS
Monsoon Winter
LEAF
A. indica Cfistula T. indica F. religiosa F. bengalensis
0.9379** 0.9043* -0.7217 0.7972
-0.5231
0.9753** 0.7167
-0.8850* 0.2512 0.9066*
0.180] 0.9432**
-0.9799** 0.8381* 0.9362**
BARK
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.6153 -0.9201** 0.9097*
-0.8757* -0.6594
0.7896 -0.3542 0.9825**
-0.8656* -0.3914
0.6013 -0.9225** 0.9801** -0.7967 -0.3736
WOOD
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.5841 -0.5668 0.4364 0.1785
-0.5817
0.5084 -0.8258* 0.4716 0.2056
-0.2368
0.6136 -0.7105 0.2781 0.0347
-0.4655
* Significant at 5% level ** Significant at 1% level
PHOSPHORUS
Phosphorus is found in the plants as a constituent of nucleic acids, phospholipids,
the coenzyme NAD and NADP and most important as a component of ATP and other high
energy compounds. Heavy concentrations of phosphorus are found in the meristematic
regions of the actively growing parts, where it is used in the synthesis of nucleo-proteins
and is also involved through ATP in the activation of amino acids for the protein synthesis.
As a constituent of nucleoproteins, it is concerned with cell division and transfer of
hereditary characters through chromosomes. As a component of phospholipids e.g.,
lecithin, phosphorus is believed to be present in the cell membranes (Devlin and Witham,
1986) Hydrogen ion (H+) carried NAD and NADP play a vital role in Kreb's cycle,
glycolysis and pentose cycle. Further, phosphate participation is directly in the
photochemical events of photosynthesis through ortho - phosphate and nicotinamide
adenine dinucleotide phosphate required for the reduction of "assimilatory power"
(Devlin and Witham, 1986). Such essential physiological processes as respiration,
nitrogen metabolism, Carbohydrate metabolism and fatty acid synthesis are all dependent
on the action of these coenzymes. Phosphorus also increases disease resistance in plants,
presumably through normal cell development resulting in vigorous growth (Tamhane et
al., 1970). Phosphorus deficiency causes decrease in the rate of protein synthesis and
results in the accumulation of carbohydrate and soluble nitrogenous compounds (Hewitt,
1963). It also causes premature leaf fall and restricted root and shoot growth.
Phosphorus is present in the soil in two general forms, organic and inorganic. It is
the inorganic form of phosphorus which is available to the plants. However, phosphorus
bound in organic compounds is eventually liberated from it through decomposition and is
released in the inorganic form that is readily taken up by the plants. Further, phosphorus
is absorbed by roots both as monovalent (H^PO/) and divalent (HPO^~) anions. The
quantity of either ion present in the soil is dependent upon pH of the soil solution. The
lower pH favours H^PO^ and higher pH, HPO^ ions (Devlin and Witham, 1986).
The observations (Table-22) regarding the concentration of phosphorus in the
foliage of different investigated species in the present study shows that in Ficus spp.
there has been an increase in the level of phosphorus under pollution stress. The
concentration of phosphorus has been the maximum in the young foliage of both
polluted and normal sets. While, in C. fistula and T. indica the concentration of
phosphorus has shown increase over the control in the young foliage, but there has
been a fall in the other seasons. However, in A. indica the winter foliage records an
increase over the control, but an opposite trend prevails in the other seasons. The
increased accumulation in phosphorus contents in the foliage under SO^ pollution
may be due to the lower rate of metabolic activity as well as the reduced growth
performance of the plants. Saxe (1983) has reported in Phaseolus vulgaris L. cv.
processor an increased concentration of phosphorus after being exposed to higher
concentration of 250 ^igm ' SO^ for 4-5 weeks. Similarly, Zech et al., (1985) have
observed increased phosphorus content in the needles of Picea abies in SO^ polluted
areas of Ne-Bavaria. The increasing trend in phosphorus concentration in polluted
plants indicates that there is no rapid transfer of minerals from leaves to other parts
such as flowers and fruits because of the reduced fruiting and flowering under
pollution stress.
The severe losses of phosphorus contents recorded in different seasons in the
rest of the species coincide with the earlier reports of many workers in different plant
forms like, in Triticum aestivum (Pandey and Rao, 1978; Prasad, 1980) Arachis
hypogea (Mishra, 1980); Pinus sativa (Garsed et ah, 1981); Cicer arietinum, Oryza
sativa, Panicum miliaceum, Vicia faha (Agrawal, 1982); Diospyros melanoxylon,
Lagerstroemia parviflora and Zizyphus nummularia (Pandey, 1983); Festuca
arundinaceae (Flagler and Youngner, 1985); Fagus sylvatica (Zech, 1990/91). The
decreased phosphorus concentrations in the plants growing under SO^ pollution in
70
the present study may be due to the inhibition in certain important enzymatic
activities'involved in 'P' metabolism. Further, it is expected that plants under stress
physiological conditions are likely to lose much energy to combat the hazards. It,
therefore, opens a challenge to the pollution problem indicating the possibility of
management of pollution stress through manipulation of T ' balance in the involved
system. Moreover, the nutrient supplying capacity of the soil with respect to P'
seems to be less (Table-4) as also reported earlier by Agrawal et al, (1985). Fried
and Broeshart (1967) put forth their hypothesis, that SO^ induced acidity may cause
greater solubility and mobility of soil constituents particularly P ' which may leach
from the rooting zone and become unavailable for plant growth.
The data further reveal that in all the investigated species in the present study
the maximum concentration of phosphorus has been observed in summer (A. indica
and Ficus spp.) or monsoon (T. indica) irrespective of treatments, with the exception
of C. fistula in which polluted leaf samples have shown the peak in summer but the
control sets recorded the same in monsoon. The decrease in concentration of
phosphorus after their peak values is perhaps due to the transfer of this mineral from
the leaves to other growing parts. Sacher (1957) and Das (1968) have also reported
that minerals are transferred to green or living parts when the leaves began to dry.
Guha and Mitchell (1966) observed seasonal variation in the concentration of
elements in leaves of deciduous trees and showed that the concentrations of elements
quantitatively decrease before leaf fall.
The correlation coefficient (Table-42) worked out in relation to leaf sulphur
concentration and the amount of phosphorus in different seasons shows that in Ficus
spp. there is a positive relationship in all the seasons and the ' r ' value ranges from 0.47
to 0.87 inF. bengalensis. In F. religiosa it shows non-significant correlation in all the
seasons. In C. fistula and T. indica, the phosphorus content of summer foliage shows
perfect positive correlation with sulphur (r = 0.93 and 0.88), but in the rest of the
71
seasons a strong negative correlation prevails
Interestingly in A. indica the concentration of phosphorus in summer and
monsoon foliage records non-significant negative correlation with sulphur (r=-0.62
and -0.75) while in the winter the correlation turns to significantly positive with sulphur
(r=0.84).
The observations (Table 23-24) regarding the concentration of phosphorous
in wood and bark show that the higher amount of phosphorus in the wood samples
of all the investigated species than bark. In C. fistula and T.indica there has been a
fall in the level of phosphorus from summer to winter in the bark and wood samples.
In A. indica an increase over the control under pollution stress has been recorded in
the bark and wtfbd, irrespective of seasons. Interestingly, F. religiosa records
increased amount of phosphorous in the bark, under SO^ pollution, but an opposite
condition prevails in the wood in all the seasons. Similarly, in F. bengalensis bark
sample records considerable depletion, while there has been a gain in the wood
samples under pollution stress.
The correlation coefficient worked out between sulphur concentration of bark
and wood and phosphorus contents in foliage reveals that the correlation is negative
in the bark and wood samples of C. fistula and T. indica in all the seasons. In C. fistula
' r ' value ranges from -0.80 to -0.89 in the bark and from -0.51 to -0.90 in wood, while
in T. indica 'r ' value shows non-significance in bark and wood. In A. indica correlation
coefficient o f 'P ' is non-significantly positive in relation to sulphur in bark as v. ell as in
wood. In Ficus species the trend is slightly different, the bark samples ofF. bengalensis
shows negative relationship with non-significant ' r ' value, but the wood shows non-
significant positive relationship with sulphur. However, inF. religiosahsirk records
positive correlation, while in the wood there has been a non significant negative
correlation.
72
Table 42. Correlation coeflicient (r) between Sulphur and Phosphorus.
SPECIES SEASONS
Summer Monsoon Winter
LEAF
A. indica -0.6289 -0.7568 0.8493* C. fistula 0.9321** -0.2774 -0.8768* T. indica 0.8886* -0.6785 -0.9792** F. religiosa 0.8478* 0.7244 0.4749 F. bengalensis 0.2386 0.5111 0.2311
BARK
A. indica 0.6254 0.5681 0.6486 Cfistula -0.8971* -0.8678* -0.8062 T. indica -0.5194 -0.3848 -0.7622 F. religiosa 0.6982 0.7213 0.5349 F. bengalensis -0.6281 -0.4000 -0.2290
WOOD
A. indica 0.6934 0.4188 0.4804 Cfistula -0.5136 -0.9338** -0.9063* T. ifidica -0.3936 -0.4096 -0.1454 F. religiosa -0.2631 -0.2096 -0.1123 F. bengalensis 0.0308 0.1067 0.0743
* Significant at 5% level ** Significant at 1% level
POTASSIUM
The potassium is required in large amounts for plants and, unlike nitrogen and
phosphorus, it does not form a stable structural part of any molecule in a plant cell.
The highest concentrations of potassium are found in the meristematic regions of the
plants. Potassium is essential for the activation of the enzymes and is involved in
the synthesis of certain peptide bonds in carbohydrate metabolism. It also enhances
the incorporation of amino acids into proteins (Webster, 1956). The enzymes that
require K as an activator include fructokinase, pyruvic acid kinase and transacetylase
(Nason and Mc Elroy, 1963). K is essential for some vital metabolic processes
including glycolysis, oxidative phosphorylation and adenine synthesis (Evans and
Sorger, 1966). It is actively involved in the translocation of solutes moving across
the sieve plates by electro-osmosis (Salisbury and Ross, 1986). Its role is inevitable
in the physiological processes like photosynthesis, chlorophyll development and the
w^ater balance of the leaves. The best known function of the K is its role in stomatal
opening and closing (Fischer and Hsiao, 1968; Humble and Hsiao, 1969). Its
deficiency causes necrosis, rosette or bushy habit of growth and weakening of stems
and decreases the resistance against pathogens.
Potassium is present in the soil in a non exchangeable (fixed) form, exchangeable
form, and in a soluble form, which always remain in equilibrium and a change in the
concentration of any one of the constituents will cause a shift towards stabilization.
For example, depletion of the soluble K in the soil by the plant and soil micro-
organisms will cause the slow release of fixed K. This situation is desirable because
absorbed and fixed K which is not readily leached from the soil can be available to
the plants.
The data regarding the concentration of potassium in the foliage of different
investigated species under pollution stress is summarized in Table-25. The depletion
in the level of potassium recorded in Ficus spp. in the present study, in different seasons
under pollution stress goes in confirmity with the earlier report of Lone (1993) in
some trees \\\it Mangifera indica. Eucalyptus citriodora, and Dalbergia sissoo from
the same site. Prasad (1980) Agrawal (1982) Zech etai, (1985) have also reported
reduced potassium content in some plants under pollution stress. The increase in
potassium content recorded in T. indica in the present study under SO^ pollution is in
agreement with the earlier reports of Saxe (1983) who found a similar increase in
potassium content in the plants exposed to SO^ pollution. However an initial increase
(43.90%) of potassium recorded in the young foliage of deciduous C. fistula may be due
to the increased accumulation ofK in this active growth stage to avoid pollution hazard,
while the significant loss (20.87%) in K content recorded under pollution stress in the
monsoon samples of A. indica maybe due to the increased depletion of potassium in this
season to combat pollution hazard due to the heavy and dense atmosphere of monsoon.
In the normal atmosphere the concentration of potassium has been recorded to the
maximum in the young foliage ofF. religiosa and the minimum in the young foliage of C.
fistula.
In the present study, the depletion of K recorded in the foliage of polluted
populations has been the highest in the winter foliage of F. bengalensis (23.75%),
and the lowest occurs in its summer foliage. It shows the utilization of more
potassium in winter to combat pollution hazard due to increased pollution and its
low disi>ersion at the site. While in summer, the young foliage accumulates more
potassium due to its transportation from older parts to newly growing regions, and
thus keeps the depletion of potassium low. In contrast to this, the maximum
percentage increase of potassium has been observed in the winter (57.78%) samples
of T. indica under pollution stress, and the lowest occurs in its young summer foliage
(0.82%). This may be due to its defence mechanism by accumulating more potassium
in this season to withstand severe stress condition. However, in the summer foliage
74
the potassium might have been utilized for other physiological processes due to its
active growth stage which probably leads to its lesser accumulation.
The correlation worked out between leaf sulphur status and potassium
concentration is summarized inTable-43. It reveals that in F;cw5 spp. the correlation
becomes negative under pollution stress because of the enhanced depletion of
potassium. Ini^. bengalemis ' r ' value falls in the range of-0.36 to -0.65, while inF.
religiosa it varies from -0.56 to -0.93. In T. indica the correlation records positive in all
the seasons, and the 'r ' value ranges from 0.22 to 0.90. In C. fistula, the summer foliage
shows perfect positive correlation with sulphur (r= 0.96), while the rest of the seasons
record negative correlation, and it becomes perfectly negative with sulphur in the
winter foliage (r=-0.91). However in A. indica only the monsoon season shows
negative correlation (r=-0.55), while it is positive in the rest of the seasons.
The observations regarding the concentration of potassium in the bark and
wood samples under polluted and normal atmosphere is abridged in Table 26-27. As
seen in leaf, the bark samples ofFicus spp. show depletion in the level of potassium
under pollution stress. While in their wood, only summer and winter samples o fF .
religiosa, and summer samples ofF. iewga/ew^/j records depletion. In A. indicah&rk
records significant depletion in the level of potassium, while the wood records an
opposite trend. In the case of C. fistula bark and wood records increased accumulation
of potassium under pollution stress except in its monsoon samples of the bark. In
T.indica the trend is slightly disturbed by recording increased amount of potassium
in bark as well as in the summer samples of the wood. Lone (1993) has reported losses
of potassium content in the bark and wood samples of Mangifera indica, Psidium
guajava, Syzygium cumini, and Eucalyptus citriodora under SO^ pollution, while he
found a significant increase of potassium content in the bark and wood samples of
Tectona grandis. However, Dalbergia sissoo recorded increase of potassium in bark
and a non-significant loss in wood in his study.
75
In general the concentration of potassium in bark shows higher than in wood
in the normal atmosphere. The depletion of 'K' content in the bark has been the
highest in the winter samples of A. indica (36.25%) and the least in the monsoon
samples of F. bengalemis (6.46%). Similarly, the increase in potassium content
under pollution records the highest in the summer samples of decidous C. fistula
(43.86%) and the least in the summer samples of T. indica (6.50%).
In wood, the maximum depletion of potassium occurs in the winter samples
of T. indica (45.51%) and the least (19.36%) in its monsoon samples. Likewise, the
increase in potassium content in wood under pollution stress occurs to the maximum
in the monsoon samples otF.religiosa (68%) and the least in the monsoon samples
of C. fistula {A.mVo).
The correlation coefficient (r) worked out between sulphur and potassium
contents in the bark and wood samples of different investigated species is given in
Table-43. The correlation becomes completely negative in all the seasons in the bark
samples of A. indica and Ficus spp. under pollution stress. While in C. fistula and
T. indica a positive correlation prevails in all the seasons with sulphur, except in the
monsoon samples of the bark of C. fistula in which the correlation is significantly
negative (r=-0.88).
In wood, A indica, C. fistula and F. bengalensis show positive correlation with
sulphur, but the correlation is non-significant in F. bengalenesis in all the seasons. In
the wood samples of T. indica and F. religiosa the trend of correlation is slightly
disturbed. The correlation becomes perfectly positive (r = 0.95) in the summer
samples of T. indica, while the other seasons record negative correlation. In F.
religiosa, the correlation is positive in monsoon, while the other seasons show
negative correlation.
76
Table 43. Correlation coefficient (r) between Sulphur and Potassium.
SPECIES
Summer
SEASONS
Monsoon Winter
LEAF
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.7633 0.9670** 0.2255
-0.6409 -0.3601
-0.5511 -0.3074 0.8624*
-0.5625 -0.6223
0.6009 -0.9130* 0.9004*
-0.9336** -0.6589
BARK
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.0467 0.9491** 0.4787
-0.7984 -0.6547
-0.5325 -0.8818* 0.6792
-0.4575 -0.9036*
-0.6062 0.4921 0.8275*
-0.3828 -0.7238
WOOD
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.2142 0.3401 0.9596** 0.0957
-0.2639
0.3413 0.3296
-0.3614 0.0893 0.3719
0.5673 0.2786
-0.2002 0.1268
-0 3989
* Significant at 5% level ** Significant at 1% level
- j o a
"1 .cs^-
S O D I U M
The sodium content ofthe earth crust is 2.8%, compared with 2.6% ofK. Most
higher plants have developed high selectivity in the uptake of K" as compared to that
of Na" , and it is particularly obvious in transport to the shoot. Plant species are
characterised as natrophilic or natrophobic depending on their growth response to
sodium and their capacity for long distance transport of sodium to the shoots. The
role of Na" in the mineral nutrition of higher plants has to be considered from two
main view points; its essentiality and/or the extent to which it can replace K" functions
in plants (Marshner, 1986)
Growth stimulation by Na^ is caused mainly by its effect on cell expansion
and on the water balance of the plant. The superiority of Na" can be demonstrated
by the expansion of sugar beet leaf segments in vivo (Marshner and Possingham,
1975) as well as in intact sugar beet plants, where leaf area, thickness, and succulence
are distinctly greater when a high proportion ofK"^ is replaced by Na" (Milford etal.,
1977). Sodium increases not only leaf area but also the number of stomata per unit
leaf area. Sodium also improves water balance of plants when the water supply is
limited. This occurs via stomatal regulation. In brief, sodium is indeed an essential
element at least for some plants, especially plants having C^ pathway. But its role
in tree species (having Cj pathway) is not known (Brownell, 1991). Literature
regarding the response of sodium under pollution stress is also very meagre.
In the present study the increased accumulation of Na" recorded in the foliage
oi Azadirachta indica, Cassia fistula and Tamarindus indica in different seasons
under pollution stress may be due to the presence of sodium salt in the ambient
atmosphere due to coal burning. Sodium sa^ is one of the by products of the
combustion of high sulphur fuel oils in thermal power plants, so its presence in the
ambient atmosphere is unavoidable (Richter et al, 1984). While a considerable
reduction in the level of Na" , recorded in the Ficus spp. under pollution stress, may
be attributed to the immediate translocation of Na" from the leaves to the other plant
parts. Westing, 1969; Mac Robbie, 1971; Bukovac and Wittver, 1957 demonstrated
this phenomenon by means of autoradiographs of bean leaves sprayed with Na".
Furthermore, the morphological character such as epicuticular waxes, cuticle, cell
wall and plasma membrane have been proposed as barriers to the absorption of solids
in general, through leaves (Logan, 1975; Hadley, 1980; Simini, 1984).
The Figure-37 on seasonal variation of Na" indicates that all the investigated
species recorded the peak amount of Na" in monsoon foliage which later recedes in
winter. This may be due to the dense and humid atmosphere of monsoon which might
have caused greater uptake of Na+ from ambient atmosphere. Swain, 1973; Logan,
1975; Simini and Leone, 1982 are of the opinion that relative humidity is an extremely
important factor with respect to sodium chloride uptake by plants. The decrease in
concentration of minerals after their peak values is perhaps due to the transfer of
minerals from the leaves to other growing parts such as flowers and fruits. Sacher
(1957) and Das (1968) have also reported that minerals are transferred to the green
or living parts when the leaves began to dry. Guha and Mitchell (1966) observed
seasonal variation in the concentration of elements in the leaves of deciduous trees
and showed that the concentration of some elements quantitatively decrease before
leaf fall. Lai and Ambasht (1982) have also observed that the decrease in the amount
of sodium in Psidium guajava in older leaves under cement dust pollution, but the
rate of decrease in polluted leaves was much slower.
The per cent variation (Fig. 40) shows that in^^. indica, C. fistula and T. indica
it becomes positive under polluted condition. The highest variation recorded in the
winter foliage of C. fistula (48%) and the lowest in the winter foliage of A. indica (6%).
Similarly, the negative variation recorded under pollution stress in the rest of the
species attains the maximum in the monsoon foliage o fF, bengalensis (31 %) and the
78
least occurs in the winter foliage of F. religiosa (16.16%).
The observations (Table 29-3 0) recorded on the Na" contents of bark and wood
in the present study show that in A. indica, C. fistula and F. bengalemis the
concentrations of Na" in bark show more than in leaves and wood in polluted
atmosphere. This is the clear evidence for translocation of sodium from leaves to
other plant parts as it has been discussed earlier in this chapter. In polluted
atmosphere indica, C. fistula and F. religiosa recorded increased amount of Na"
in bark, while the wood records considerable loss of Na"* in this species. In T. indica
bark recorded a non-significant loss in the amount of sodium and at the same there
has been a non-significant gain in wood. F. bengalensis is the only species in the present
study which shows loss of Na" in leaves, bark as well as in wood under pollution stress.
Since, the sodium has not been considered as an essential element for Cj species, the
sodium involved biochemical processes in this species are not known at present. The
increase in the amount of sodium recorded in bark happens to be the maximum in the
monsoon season of C. fistula (82.05%) and the minimum in the monsoon samples of
A. indica (15.3%%). Similarly, the loss ofNa^ under pollution stress being the highest
in the winter samples ofF. bengalensis (45.45%) and the lowest in the winter samples
of the bark of T. indica (20%). In wood the loss of sodium under pollution stress occurs
to the maximum in the winter samples oi C. fistula (44%) and the minimum in the
winter samples ofF. religiosa {MVo).
The correlation (Table-44) worked out between leaf sulphur statu'' and
sodium shows that the correlation is completely positive in different seasons in C.
fistula, but it is significant only in winter samples (<0.05%). Similarly, F. religiosa
records negative correlation in all the seasons with sulphur and its summer and
monsoon samples only show significant correlation (<0.05%). The rest of the species
records negative and positive correlation in different seasons, and it comes to the
level of significance only in the summer foliage ofT.indica (<0.05%).
79
In bark (Table-44), A. indica, C. fistula and F. rehgiosa show positive
correlation with sulphur C. fistula shows significant correlation in all the seasons,
while the rest of i c species records non-significant correlation except in the winter
samples of F. religwsa However, T. indica and F. bengalensis show negative
correlation with sulphur except in the winter samples oiF. bengalensis in which the
correlation records positive The significant correlation (<0 05%) occurs only in the
monsoon samples of T. indica
Likewise, in wood (Table-44)y4. indicannA C./w/w/a record negative correlation
with sulphur, which has been found significant only in the monsoon samples of
C.fistula (<0.05%), while the rest of the species record non significant positive and
negative correlation in different seasons.
The correlation (Table-45) worked out between potassium and sodium in
leaves shows that the correlation is positive in F. religwsa in all the seasons. While
the rest of the species records positive and negative correlation in different seasons
depending on the extent to which it replaces K" functions. But none of the correlation
in leaves with potassium has shown statistical significance.
In bark (Table-45), A. indica, T. indica and F. religiosa show negative
correlation with potassium. But it goes to the level of significance only in the winter
samples o{A. indica and T. indica and in the monsoon samples o f F . religiosa The
positive correlation recorded with potassium in the rest of the species is non-
significant in different seasons
In wood (Table-45), indica, C.fistula, T. indica and F. bengalensis shovii
negative correlation with potassium except in the summer samples of T.indica and
F. bengalensis But it shows significant correlation only in the winter samples of C.
fistula and in the monsoon samples of T. indica and F. bengalensis In F. religiosa,
the correlation shows a non-significant positive in summer and winter samples, and
a non-significant negative correlation in monsoon
80
Table 44. Correlation coefllcient(r) between Sulphur and Sodium.
SPECIES
Summer
SEASONS
Monsoon Winter
LEAF
A. indica C. fistula T. indica F.religiosa F. bengalensis
0.6747 0.3147 0.8563* -0.8896* -0.7907
-0.7406 0.6761 0.7804
-0.8281* 0.5988
0.4159 0.8846*
-0.2218 -0.4151 -0.6479
BARK
A. indica Cflstula T. indica F. religiosa F. bengalensis
0.6331 0.8833*
-0.8067 0.6559
-0.5921
0.5919 0.8937*
-0.8309* 0.7900
-0.7413
0.6142 0.9083* 0.8100 0.8514* 0.4938
WOOD
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.6075 -0.3333 0.2918
-0.4607 0.0105
-0.5438 -0.8747* 0.5051 0.0855 0.0552
-0.6585 -0.7370 -0.5012 0.1313 0.2474
* Significant at 5% level ** Significant at 1% level
Table 45. Correlation coefficient (r) between Potassium and Sodium.
SPECIES
Summer
SEASONS
Monsoon Winter
LEAF
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.7115 0.5042 0.2704 0.3333 0.5339
-0.5585 -0.4889 0.4378
-0.1106 -0.6943
0.4562 -0.7911 -0.2506 0.5198 0.6940
BARK
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.5943 0.6282
-0.5937 -0.6869 0.5536
-0.4367 0.6581
-0.7018 -0.8868* 0.7701
-0.8768* 0.4492
-0.8627* -0.7786 0.7118
WOOD
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.7044 -0.4130 0.6435 0.6428 0.2331
-0.3508 -0.4086 -0.8142* -0.6089 -0.8193*
-0.7387 -0.8340* -0.3894 0.5242
-0.7638
* Significant at 5% level ** Significant at 1% level
C A L C I U M
Calcium is the major exchange cation offertile soils. It is absorbed as divalent
. However, the major portion of calcium in the soil is found in a non-exchangeable
form, chemically bound in primary minerals such as anorthite (Ca Al Si^Og). Calcite
(CaCOj) is present in soils of semi arid and arid regions, and insoluble calcium
phosphate salts occur in alkaline soils. Some of these calcium salts are available to
the plant, depending on the solubility and the degree of alkalinity of the medium
(Devlin and Witham, 1986).
One well-known role played by calcium in the plant is that if forms a
constituent of cell walls in the form of calcium pectate which forms the middle
lamella, composed primarily of calcium and magnesium pectates. Calcium is thought
to be important in the formation of cell membranes and lipid structures. Most calcium
in plants is in central vacuoles and bound in cell walls to pectate polysaccharides
(Kinzel, 1989). Calcium in small amounts is necessary for mitosis. In this respect,
Hewitt (1963) has suggested that calcium may be involved in chromatin or mitotic
spindle organization. Abnormal mitosis may develop due to calcium deficiency on
chromosome structure and stability. Calcium may also be an activator for the enzyme
arginine kinase, adenosine triphosphate, adenyl kinase, and potato apyrase (Mazia,
1954).
The easily observed symptoms of calcium deficiency are striking. Meristematic
regions found at stem, leaf, and root tips are greatly affected and eventually die, thus
terminating growth in these organs. Malformation or distortion of the younger leaves
is also characteristic of calcium deficient plants, a hooking of the leaf tip being the
most easily detected symptom. Deficiency symptoms appear first in the younger
leaves and in the growing apices, probably as a consequence of the immobility of
calcium in the plant (Kirkby and Pilbeam, 1984).
The observations (Table-31) recorded on the calcium content in the foliage
of different investigated species in the present study show that there has been an
increase in the level ofcalcium in the foliage ofi4. indica and T. /W/ca under pollution
stress. Several workers in the past have also reported increase in calcium content in
the foliage of different plant species under pollution stress (Materna, 1961; Malkonen,
1974; Garsed et al., 1981; Amundson et al, 1990/91). However, the decrease in
calcium content under pollution stress recorded in the foliage of C. fistula and Ficus
spp. in the present study in different seasons goes in agreement with the earlier report
of Zech et al, (1985) in some forest tress of SO^ polluted areas of Ne-Bavaria and
Jacobson et al., (1989) in Picea rubens (Red spruce) when exposed to acid mist
containing major sulphur and nitrogen pollutants. Wookey and Ineson (1991) found
calcium depletion in Pinus sylvestris during the open air fumigation with SO^.
Schatzle et al, (1990) have also reported a decreasing trend in calcium content in the
needles of Fir (Abies alba) and Spruce {Picea abies) due to SO^ treatment. In the
normal atmosphere the concentration of calcium has been found to the maximum in
the monsoon samples of F. religiosa and the minimum in the monsoon samples of
A. indica. The enhanced depletion of calcium under pollution stress occurs in the
winter foliage ofF. bengalensis {\9.OIV^,) and the least occurs in the summer foliage
of F. religiosa (4.20%). Similarly, the increase in calcium content in foliage under
polluted condition has been the maximum in the winter foliage of T. indica (10.40%)
and the least in its summer foliage (7.79%).
The correlation (Table-46) worked out between leaf sulphur content and
calcium concentration shows that indica and T. indica have a positive relationship
with sulphur in the calcium content of their foliage under pollution stress. The 'r '
value falls in the range ofO.86 to 0.93 '\r\A. indica, 0.78 to 0.93 in T. /W/ca in different
seasons. However, in the rest of the species worked out, the ' r' value shows negative
correlation with sulphur in the different seasons.
8 2
The observations (Table 32-33) on bark and wood show that the different
investigated species posses higher capacity to accumulate much more calcium in their
bark than in leaves and wood in normal as well as polluted conditions. In C. fistula,
bark and wood accumulate significantly elevated amount of calcium under pollution
stress, while in^^. indica, T. Jndica and F. religiosa hiirV. samples record increased
accumulation of calcium, but the wood shows a negative trend in all the three. In F.
bengalensis bark and wood accumulate significantly high amount of calcium, except
in winter samples of wood, in which there has been a fall. In general, increased calcium
accumulation takes place in the bark samples of different investigated species and
it has been the maximum in the monsoon samples of J. m<i;ca (18 .38%) and the least
in the older barks of F. bengalensis (2.28%). Similarly, in the wood samples, the
maximum increase has been recorded in the winter samples of C. fistula and the least
in the summer samples ofF. bengalensis (3.36%). The depletion in calcium content
recorded over the control has been the maximum in the winter samples of C. fistula
(28.78%) and the minimum in the winter samples of F. religiosa (3.03%).
The correlation coefficient (Table-46) worked out between sulphur
concentration and the level of calcium in the bark and wood shows that the correlation
is positve in the bark samples of all the species, except F. religiosa in which the
samples of monsoon and winter seasons record negative correlation. The correlation
is significant in the summer and winter samples of C. fistula and in the winter samples
of T. indica and F. religiosa. The wood samples of A. indica, T. indica and F.
religiosa show negative relationship with sulphur in their calcium concentration,
while in C. fistula and F. bengalensis a positive correlation with sulphur exists except
in the winter samples of the wood of F. bengalensis, where the correlation is negative.
83
Table 46. Correlation coefHcient (r) between Sulphur and Calcium.
SPECIES
Summer
SEASONS
Monsoon Winter
LEAF
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.8631* -0.8894* 0.9356**
-0.7718 -0.8176*
0.9311 -0.9305 0.8997
-0.9512 -0.9554
0.9211 -0.9512 0.7868
-0.9223 -0.8516
BARK
A. indica C. fistula T. indica F. religiosa F. bengalensis
0.8091 0.9166* 0.3361 0.0883 0.7108
0.7344 0.6032 0.7396
-0.1859 0.3347
0.7244 0.9143* 0.8544*
-0.8285* 0.4752
WOOD
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.5897 0.8654*
-0.3306 0.2686
-0.3844
0.6629 0.8572* 0.3429 0.2220
-0.2586
-0.7426 0.7936
-0.5201 -0.2143 -0.0185
* Significant at 5% level ** Significant at 1% level
ASCORBIC ACID
Ascorbic acid otherwise known as vitamin C is an antiscorbutic. It is a water
soluble and heat-labile vitamin. According to Keller and Schwager (1977), continued
exposure of plants to SO^ reduces ascorbic acid contents long before the appearance of
visible injury symptoms. Since ascorbic acid acts as a strong reductant and it is
responsible for photoreduction of protochlorophyllid, its decrease may lead to several
physiological complications in plants (Rudolph and Bukatsch, 1966). The reduction in
level of ascorbic acid in pollutant exposed plants has been ascribed to enzyme toxicity and
sulphonation of S-H groups (M^pson, 1958). Ballantyne (1973) has shown that
exposure to SO^ alters the ratio of oxidized to reduced sulphydryl groups in plants.
Hogler and Herman (1973) and Young and Loewus (1975) suggested that in the presence
of SOj, the ascorbic acid might be reduced or converted into Dehydro ascorbate (DHA),
or oxalic acid/or into other convertible carbohydrates. Chaudhary and Rao (1977)
related pollution tolerance of plants with their ascorbic acid levels and concluded that the
higher the level of ascorbic acid the greater the tolerance.
In the present study all the investigated species recorded decreased level of
ascorbic acid in their foliage under pollution stress irrespective of seasons (Table-
34). Rao and Dubey (1990a) found similar observations in Cassia siamea, Dalbergia
sissoo, Calotropisprocera and Ipomeafistulosa, growing at different low levels of
ambient air pollution in Ujjain (India). Sharma and Rao (1985) observed increase in
ascorbic acid in SO^, HF and SO^+HF fumigated plants at initial stage, followed by
decrease at 55 day plant age. Kamalakar (1992) has reported reduced ascorbic acid
content in some tropical species under automobile pollution. Sharma and Prakash
(1991) observed significant reduction in ascorbic acid in leaves upto 48.95% in 1334
kg m ' SOj in 60 days old plants ofLycopersicon esculentum. Varshney and Varshney
(1984) also reported the effects of SO^ on ascorbic acid content oiBrassica nigra,
Phaseolusradiatus and Zea mays. The latter is relatively SO^ resistant and possesses
a comparatively high amount of ascorbic acid. The ascorbic acid contents ofB. nigra
and P. radiatus, which are relatively SO^ sensitive species, decreased markedly
within one week following SO^ fumigation. This reduction is due to the oxidizing
property of the sulphur dioxide. Sulphur dioxide readily diffuses into mesophyll
cells of leaves and produces oxy-radicals. These oxy-radicals reduce ascorbic acid
to dehydro ascorbic acid. The decrease in amount of ascorbic acid makes the plant
more sensitive to SO^.
In the present investigation, in normal as well as SO^ enriched conditions the
highest concentrations of ascorbic acid occurs in the foliage of deciduous Cassia
fistula. This higher concentration of ascorbic acid in their foliage makes the plant
more resistant to SO^ pollution. While in Ficus spp. the level of ascorbic acid in
normal as well as polluted atmosphere has been recorded low, this resulted in greater
reduction of this premier vitamin under SO^ enriched condition in this species. This
has been directly correlated with increased fixation of sulphate-sulphur in these two
species. In T. indica the level of ascorbic acid slightly improves than in Ficus spp.
in both the sets in different seasons. It records a more or less 38% reduction of
ascorbic acid in different seasons. A. indica in the present study records elevated
amounts of ascorbic acid in various seasons, in treated as well as control populations.
As seen in C. fistula, this species also shows tolerance to SO^ pollution due to its
increased level of ascorbic acid. The reduction varies 21.36% to 31.27% in this
species in different seasons. The conclusion of Chaudhary and Rao (1977), the higher
the level of ascorbic acid the greater the tolerance in plants.
However, in the present investigation the high reductions of ascorbic acid
occur in the young foliage of summer season in all the investigated species under
pollution stress. This may be due to the easy sorption of SO^ in the young foliage.
The increased concentration of ascorbic acid recorded in winter season, in the foliage
85
of control and polluted populations of all the species studied, may be due to the
increased SO^ pollution coupled with increased water stress at the sites due to cold
and dry climate, which might have resulted increased production of ascorbic acid.
Hunter et al., (1950) are able to demonstrate an appreciable enhancement of
ascorbic acid content in the upper leaves ofBrassica rapa plants grown under low soil
water potential. Other workers have also reported increased ascorbic acid
concentration in plants during a period of severe water stress (Solomon, 1955;
Novitskaya, 1958). Similarly, Farkes and Rajhathy (1955) found an enhanced
concentration of ascorbic acid in tomato plants grown under dry conditions.
The correlation (TabIe-47) worked out on sulphur versus ascorbic acid
production in the foliage of different investigated species in the present study show
that, there is a negative relationship between sulphur and ascorbic acid in different
seasons. In C. fistula and F. religiosa the correlation records perfectly negative
(p<0.01% level) in all the seasons. While in the rest of the species the negative
correlation coefficient (r) shows p<0.01% to p<0.05% level significance in various
seasons.
86
Table 47. Correlation coefTicient (r) between Sulphur and Ascorbic Acid in the foliage.
SPECIES
Summer
SEASONS
Monsoon Winter
A. indica
C. fistula
T. indica
F. religiosa
F. bengalensis
-0.8544*
-0.9894**
-0.8889*
-0.9441**
-0.9056*
-0.9353**
-0.9788**
-0.8941*
-0.9635**
-0.9196**
-0.8962*
-0.9903**
-0.9908**
-0.9562**
-0.9249**
•Significant at 5% level ** Significant at 1% level
PROLINE
Proline is a basic amino acid found in high percentage in basic proteins. Free
proline in plants is said to play a role under stress conditions. Though the molecular
mechanism has not yet been established for the increased level of proline, one of the
hypothesis refers to breakdown of protein, into amino acids and conversion to
proline for storage (Sadasivam and Manikam, 1992). An increase in free proline
content occurs in plants subjected to the action of such harmful factors as dryness,
salinity or air pollution, which disturb their water balance. Investigations carried out
by many workers reveal that the accumulation of free proline may be utilized as
sensitive biochemical indicator of water stress in plants, whether under influence of
drought (Bode et al., 1985), salinity (Buhl and Stewart, 1983) or toxic gases
(Karolewski, 1985). In a polluted environment, plants are frequently exposed to the
simultaneous action of many factors which promote water stress.
In the present study (See table-35) there has been an increased production of
free proline in the foliage of C.//5/w/a, T. indica&nAFicussiip. under pollution stress,
irrespective of seasons. Several workers in the past have recorded increased
accumulation of free proline in different forms of plants under varied stress
conditions, which disturbs their water balance (Boggese^a/., 1976; Karolewski, 1984
a, b, 1989; Bode et al., 1985; Kamalakar, 1992). In the present study the highest
percentage (67.80%) of proline production over the control has been recorded in the
young foliage of the polluted samples of C. fistula in the summer season. Norby and
Kozlowaski (1981) have reported increased production of free proline in SO^ polluted
plants under high temperature. They interpreted it may be due to the greater SO^
uptake in high temperature. In high temperature the diffusive conductivity of the
foliage also increase. The more rapid penetration of SO^ into the leaves, together
with an increase of temperature, was also reported by Heck and Dunning (1978).
However, in the present study the concentration of free proline has been the maximum in
the winter samples in all the species, irrespective of treatments. The mechanism of free
proline accumulation, caused by the action of low temperature may be similar to that
which operates on exposure to SO^ (Karolewski, 1989). Increase in the content of this
imino acid under the influence of SO^ and low temperature has also been observed by
Tesche (1979) in Norway Spruce. The action of SO^ on plant causes the release of large
quantities of toxic ammonia (Godzik and Linskens, 1974) and its reaction with a -
ketoglutaric acid, together with the formation of the intermediate glutamate (Jager and
Pahlich, 1972), is generally believed to be the pathway of proline synthesis, and of the
formation of amino acids in plants following SO^ action (Malhotra and Sarkar, 1979).
This constitutes a defence mechanism against the toxic action of this gas. The significant
reduction in the proline content recorded in the foliage of A. indica'xn all the seasons may
be directly related to the increased synthesisofprotein under pollution stress. The break
down of protein into amino acids and conversion to proline for storage under stress
conditions forms one of the explanation for increased proline. However, C. fistula and
Ficus spp. exhibit increased protein as well as proline in their foliage in different seasons
under pollution stress. In the normal and polluted conditions the concentration of proline
has been the maximum in the winter samples of A. indica and the least in the monsoon
samples of F. bengalensis. This makes the role of proline more clear, that in winter
the plant populations are experiencing more stress because of cold and dry climate
and thus producing more proline, while in monsoon the plants are receiving sufficient
amount of water because of rain and thus there is least chance for water stress which
does not warrant high level of free proline. Karolewski (1989) has found that the
simultaneous action of more than one factor had a greater influence on proline
production than any single factor alone. Further, he interprets that the defence
mechanism of the plant is manifested by an increase in the level of this imino acid
and it appears to play a detoxification role, with the possibility of its later utilization
in regeneration process
8 8
The correlation coefTicient (Table-48) worked out between leaf sulphur content
and proline production shows that the correlation is positive in all the species except in
A. indica. C. fistula And T. m<//ca record significant correlation in all the seasons. In
Ficus spp., the winter season of F. religiosa as well as summer and monsoon seasons
of F. bengalensis shows significant positive correlation with sulphur. However, in A.
indica all the seasons record perfect negative correlation with sulphur (r=-0.89 to -0.95).
The correlation (TabIe-48) worked out between leaf protein concentration and
proline show that in A. indica and T. indica there has been a negative correlation
between protein and proline in all the seasons. In the rest of the species the
correlation records positive relationship with protein except in the summer samples
of the foliage of F. bengalensis. However, none of the correlation show statistical
significance with protein with the exception of the summer samples of A. indica in
which correlation goes to the level of significance (p<0.05%).
89
Table 48. Correlation coeflicient (r) between Sulphur, Proline and Protein in the foliage.
SPECIES
Summer
SEASONS
Monsoon Winter
Sulphur and Proline
A. indica Cfistula T. indica F. religiosa F. bengalensis
-0.9184** 0.9502** 0.8312* 0.6194 0.7933
0.9509** 0.8339* 0.8972* 0.8265* 0.8677*
-0.9472** 0.9531** 0.8459* 0.9645** 0.5570
Protein and Proline
A. indica C. fistula T. indica F. religiosa F. bengalensis
-0.8916* 0.7714
-Q3122 0.2662
-0.1133
-0.7314 0.2451
-0.4869 0.4717 0.7430
-0.2428 0.6763
-0.1568 0.7449 0.4972
•Significant at 5% level ** Significant at 1% level
07 S'LL'LCrE'D STT.CI'ES
COMPARATIVE PERFORMANCE OF THE
SELECTED SPECIES
The overall assessment of the different parameters taken into account in the
present study indicates that Azadirachta indica possesses high level of free proline
in the leaves and a moderate level of ascorbic acid showing that this species is
comparatively more tolerant to coal-smoke pollution among the investigated forms.
This is supported by high protein synthesis, high level of nitrogen and also high
amount of sulphur accumulation in leaves. The high level of protein synthesis and
nitrogen rich condition obviously shov^ the species is performing well under sulphur
enriched environment. The high percentage of sulphur accumulation under pollution
stress indicates the capability of the species to convert the harmful form of sulphur
in its oxide form which is a harmless inorganic sulphur compound suitable for
storage. A similar analysis of Cassia fistula and its performance under coal smoke
pollution stress shows this is another species which could withstand high load of
pollution by having high amount of ascorbic acid and a high amount of free proline.
Here also high protein synthesis, high level of nitrogen and rich amount of carbohy-
drate corroborate the assumption that Cassia fistula is another suitable form with
high degree of tolerance to coal smoke pollution. In this species as in A. indica, the
sulphur accumulation is high. Tamarindus indica on the other hand, loses coloured
pigments, carbohydrates, proteins, nitrogen and phosphorus to a significant level but
accumulates free proline to a considerable extent and to a moderate level of ascorbic
acid indicating its ability for survival under coal smoke stress. The broad leaved
Ficus bengalensis, the true tropical form, performs well under coal-smoke pollution,
although it has poor level of free proline and ascorbic acid. Undoubtedly this species
has the high capability to fix large amount of sulphur and at the same time showing
significant increase in carbohydrate synthesis owing to enhanced chlorophyll level
and rich amount of protein, nitrogen and phosphorus. This is the only species which
accumulate rich amount of calcium and potassium. The other Ficus species viz. Ficus
religiosa also performs well but to a lesser extent as compared to Ficus bengalensis,
although it fixes the highest amount of sulphur under coal-smoke pollution.
91
C0ih^LUSI09i
CONCLUSION
The overall assessment of the different biochemical parameters taken into
account to study the effect of coal smoke pollution in the present investigation on A.
indica, C. fistula, T. indica, F. beugalensis md F. religiosa, unreels the following:-
a. The different species respond differently to coal-smoke pollution depending
on its own genetic constitution and in the climatic condition as well as the part
or organ of the concerned plant and, therefore, it becomes inevitable for the
investigator to study the individual speices as well as the different parts of an
individual plant before anything could be said definite about the behaviour of
the species as a whole.
b. The coal-smoke pollution causes an increase in the sulphur content in leaves,
bark and wood of all the species irrespective of the seasonal conditions
prevailing in the ambient atmosphere.
c. The coal-smoke pollution causes severe destruction of coloured pigments in
the foliage of all the investigated species wdth the single exception of F.
bengalensis in which the pigment strategy has been fotmd to be highly
productive. Chlorophyll-b undergoes a greater loss in A. indica and F.
religiosa, while in the rest chlorophyll-a suffers high percentage of loss.
d. The amount of carbohydrate loss or gain under pollution load is more
pronounced in wood and bark than in the foliage except in F. bengalemis in which
the maximum loss of carbohydrate has been found in leaves.
e. Protein synthesis gets enhanced in the foliage of all the investigated species
under ambient pollution load except in T. indica. The bark and wood show
different trends. The magnitude of the variation depends on the seasonal
conditions, part of the plant as well as the genetic constitution of the species
concerned.
f. The nitrogen accumulation under coal smoke pollution follows the same trend
as in protein, in the foliage, bark and wood of all the investigated species.
g. The phosphorus concentration undergoes a declining trend under coal smoke
pollution in the various parts of T. indica, whereas in the rest of the species,
the pattern varies in different parts depending on the growth pattern of the
concerned species.
h. The coal-smoke pollution causes severe depletion of potassium in Ficus spp.,
while in others, it elevates the amount in foliage. In the bark and wood it
causes increase or decrease depending on seasonal conditions.
i. The sodium content shows altered rhythms in the foliage, bark and wood of
all species studied under the stress of coal-smoke pollution. The degree of
uptake or accumulation depends on the inherent behaviour of the species as
well as the status of potassium.
j. In the normal as well as polluted atmosphere, the concentration of calcium
shows its peak in the bark samples of all the species investigated. The amount
of calcium shows marginal variation in the leaves, bark and wood of all the
species under pollution stress, depending on the species as well as the
meteorological condition predominating in and around the sites of study.
k. Pollution causes a severe reduction in ascorbic acid content in the foliage of
all the species in different seasons. High concentration of ascorbic acid in the
foliage has been found in A. indica and C. fistula, both under normal as well
as under the pollution load, compared to other species investigated.
1. The ambient pollution load leads to the enhanced production of free proline
in the foliage of all the species studied in various seasons, except \nA. indica
which records a decreasing trend.
9 3
SUMMARY
The effect of air pollutants emerging out of coal-burning at the Thermal Power
Plant Complex, Kasimpur, Aligarh, has been studied on the biochemical responses of
some perennial broad-leaved forms of tropical origin namely, Azadirachia indica
(Meliaceae), Cassiafistula (Caesalpiniaceae), 7a/nar/wt/tt5'/W/ca(Caesalpiniaceae),
Ficus religiosa (Moraceae) and Ficus hengalensis (Moraceae). The data were
collected on the level of sulphur, photosynthetic pigments, carbohydrate, protein,
nitrogen, phosphorus, potassium, sodium, calcium, ascorbic acid and proline in the
foliage, newly formed bark and sap wood samples.
Sulphur
The sulphate-sulphur content shows highly significant accumulation of sulphur
in foliage under pollution stress. The peak amount of sulphur occurs in the monsoon
foliage in all the species with the exception of Cassia fistula. The magnitude of
sulphate-sulphur accumulation depends on the season as well as the inherent behaviour
of the species concerned. The accumulation of sulphate-sulphur in decreasing order
in the foliage falls as, Ficus religiosa < Ficus hengalensis < Cassia fistula,
Azadirachia indica < Tamarindus indica. Similarly in bark, significant amount of
sulphur accumulates except in T. indica. The highest percentage of sulphur accumu-
lation in bark takes place inF. hengalensis and the least in A. indica. In wood, also
significant amount of accumulation of sulphate-sulphur takes place under coal-smoke
pollution.
Chlorophyll and Carotenoids
The photosynthetic pigments show severe losses in their amount in all the
investigated species, except inF. in which they increase. Chlorophyll 'a '
suffers greater loss in C. fistula and T. indica, while in the rest chlorophyll 'b'
undergoes greater loss under coal-smoke pollution. The peak amount of photo-
synthetic pigments has been found in the monsoon foliage in all the species irrespec-
tive of pollution level. The ratio of chlorophyll 'a ' to chlorophyll 'b' ranges from
1:0.61 to 1:0.77 under pollution. The carotenoid contents also shows decreasing trend
under pollution. The ratio of carotenoids to chlorophyll varies from 1:0.22 to 1:0.26
in the control and from 1:0.18 to 1:0.26 in polluted populations. The correlation of
photosynthetic pigments with sulphur shows negative relationship in all the species
except in F. hengalensis, where the correlation turns out to be positive.
Carbohydrate
Under coal-smoke pollution, all the species have recorded increase in carbo-
hydrate in the foliage, except in T. indica. In A. indica, C. fistula mAF. hengalensis
there has been a steady increase of carbohydrate from younger to older foliage. The
increase in carbohydrate under pollution stress has been the maximum in the foliage
ofF. hengalensis, followed by^ . indica, C. fistula andF. religiosa. The amount of
loss or gain under coal-smoke pollution has been more pronounced in wood than in
bark and the foliage. In bark, all the species have recorded gradual increase in
carbohydrate from summer to winter except in C. fistula. The bark samples of A.
indica, C. fistula and F. religiosa record increased amount of carbohydrate under
pollution. The concentration of carbohydrate is higher in wood than in the foliage and
bark in the normal as well as polluted atmosphere. All the species have recorded
decreased level of carbohydrate in wood under coal-smoke pollution except in F.
hengalensis. The correlation of carbohydrate with sulphur shows, negative as well
as positive relationship in different species in various season, in the foliage, bark and
wood. Similarly, the correlation with photosynthetic pigments shows varied pattern
in different species.
95
Protein
The protein content shows increasing trend under coal-smoke pollution in all
the investigated species except in T. indica. However, the significant interaction has
been found only in F. bengalensis. The concentration of protein showed varied trend
in different species in various season. In bark, the decreased level of protein has been
recorded under pollution stress in C. fistula and F/cwJspp.. The seasonal variation
in bark has been found to be non-significant in A. indica, C. fistula and Ficus spp..
In wood, A. indica, T. indica and F. religiosa record increase in the amount of protein,
while the rest show decreased level of protein under coal-smoke pollution. The
highest increase in protein level in wood under pollution occurs in the winter samples
of A. indica while the loss attains the maximum in the summer samples of F.
bengalensis. The correlation of protein with sulphur in the foliage shows mostly
positive relationship, with varying level of significance. In bark and wood it shows
negative as well as positive relationship in different species.
Nitrogen
The nitrogen content in the foliage follow the same trend as described in case
of protein under coal-smoke pollution. The investigated species have recorded
increased accumulation of nitrogen in the foliage under pollution except in T. indica.
In A. indica and F. religiosa there has been a decrease in nitrogen content from
younger to older foliage. The accumulation of nitrogen under pollution stress in
decreasing order falls as, A. indica, F. bengalensis, F. religiosa and C. fistula. In
bark, significant accumulation of nitrogen under coal-smoke pollution has been
recorded in A. indica and T. indica, while in the rest nitrogen level decreased
compared to control. In wood, A. indica, T. indica and F. religiosa indicate increased
nitrogen content under coal-smoke pollution, while the rest of the species show
decreased amount under similar condition. The seasonal trend in nitrogen accumula-
96
tion also varies in different species in wood. The correlation of nitrogen content with
sulphur in the foliage shows positive relationship in all the species with the exception
of T. indica in which the correlation is negative. In bark and wood, A. indica and T.
indica show positive correlation in different season, while in the rest of the species,
the pattern of correlation varies.
Phosphorus
The phosphorus content show increasing trend under coal-smoke pollution in
the foliage oiFicus spp., while in the rest there has been a significant loss. T. indica
did not show any change statistically under coal-smoke pollution. In all the species
studied the peak level of phosphorus is recorded in summer except in T. indica which
has got a monsoon peak. This vital element experiences severe depletion in the bark
samples of C.//\s/M/a, T. indica, andF. while the rest of the species show
gain in their phosphorus content under pollution. The concentration of phosphorus
increases from summer to winter in the samples of bark in all species except in A.
indica and F. bengalensis. The sap wood samples show higher concentration of
phosphorus than in the bark A. indica and F. bengalensis show significant increase
in the phosphorus content in wood under coal-smoke pollution, while the rest record
significant loss. The correlation of phosphorus with sulphur content in the foliage
shows positive as well as negative relationship in different species. The trend has
been almost similar in bark and wood.
Potassium
The potassium content in the foliage of Ficus spp. experiences significant
depletion under coal-smoke pollution, while in T. indica X \ \ Q X Q has been a significant
accumulation. The rest of the species show non-significant variation. In the polluted
atmosphere T. indica has shown the maximum ability to accumalate potassium in
97
foliage and the minimum is shown by A. indica. In C. fistula and Ficus spp. the
maximum per cent variation in potassium under coal-smoke pollution has been
observed in winter. In bark the concentration of potassium shows higher than in wood.
A. indica and Ficus spp. show significant depletion in the level of potassium in bark
under coal-smoke pollution, while T. indica has shown increase as in the case of
leaves. A. indica and F. bengalensis record the peak amount of potassium in the
monsoon samples irrespective of treatments. C. fistula shows a unique behaviour by
recording positive variation in potassium in summer and winter, and a significant
negative variation in monsoon. In wood the investigated species show increased as
well as decreased level of potassium content under coal-smoke pollution. The
maximum depletion in potassium content in wood under coal-smoke has been shown
by the monsoon samples of F. religiosa, and the maximum gain is recorded in the
summer samples of T. indica. The correlation of potassium with sulphur shows
positive as well as negative relationship in the leaves, bark and wood samples of
different species.
Sodium
iiic loiiage of A. indica, C. fistula and i. indica recuius c^ignincant increase
in sodium content under coal-smoke pollution, while in the rest of the species there
has been a significant loss. The peak amount of sodium accumulation has been
recorded in the monsoon foliage of all the specie", but the interseasonal variation
happens to be non-significant in^. indica andF. religiosa. The maximum accumula-
tion of sodium occurs in the winter foliage of C. fistula over the control, and the
maximum loss is experienced in the monsoon foliage of F. bengalensis. The sodium
. content in bark shows different pattern under pollution. Iny4. indica and F. religiosa
the peak concentration of sodium is record in winter, while in the rest the same
happens in other seasons. The maximum gain in sodium in bark under coal-smoke
98
pollution occurs in the monsoon samples of C. fistula and the maximum loss occurs
in the winter samples of F. bengalensis. All the species show decreased level of
sodium in wood under pollution except T. indica. The maximum loss occurs in the
winter samples of C. fistula. The correlation of sodium with sulphur shows negative
and positive relationship in different species. The correlation of sodium with
potassium depicts non-significant correlation in the foliage, but bark and wood show
varied trend.
Calcium
The calcium level in the foliage shows reduction under coal-smoke pollution
in C. fistula and Ficus spp., while in the rest of the species it increases. In Ficus spp.
the peak level of calcium in the foliage occurs in monsoon, while in the rest the same
is record in other seasons. In A. indica and T. indica it records a non-significant fall
in calcium concentration from summer to winter. The per cent variation noted in C.
fistula and F. bengalensis shows significance under pollution stress, while it has
been non-significant in the other species. The bark shows the maximum ability to
accumulate calcium compared to the foliage and wood in normal as well as polluted
atmosphere. The level of calcium increases in the bark samples of all the species
under coal-smoke pollution, except in F. religiosa. The level of calcium shows
gradual increase from summer to winter in all the species. In wood, C. fistula and F.
bengalensis show increasing amount under pollution, while the rest show declining
trend. The significant variation of calcium in wood under pollution occurs only in T.
indica. The correlation of calcium with sulphur shows positive relationship in A.
indica and T. indica in the foliage, while the rest show negative relationship. In bark,
all the species show positive relationship with sulphur except in F. religiosa, while
wood shows a different trend.
99
Ascorbic acid
The ascorbic acid content shows a decreasing trend under pollution in all the
species. The concentration of ascorbic acid in the foliage increases with the age of
the leaves. The level of ascorbic acid remains statistically same in C. fistula under
coal-smoke pollution. In A. indica, T. indica znA Ficus spp. as higher loss of ascorbic
acid occurs in summer foliage, the degree of loss in ascorbic acid under coal-smoke
pollution in decreasing order falls as, F. religiosa < F. bengalensis < T. indica <
A. indica < C. fistula. The correlation of ascorbic acid with sulphur shows significant
negative correlation in all the species in different seasons.
Proline
The content increases under coal-smoke pollution in all the species except in
A. indica. The peak level of free proline records in the winter foliage of all the species
after a sharp fall in monsoon, irrespective of treatments. The per cent variation shows
maximum and minimum in different seasons. The accumulation of free proline in
increasing order under pollution falls as, F. bengalensis > F. religiosa > T. indica
> C. fistula.
The correlation of proline with sulphur shows positive relationship in all the
species except in A. indica in which it is negative. The correlation of proline and
protein shows negative as well as positive correlation in different seasons. Only the
summer samples of A. indica shows significant correlation with protein.
The overall assessment of the parameters undertaken in the present study has
revealed that all the investigated species are good enough for plantation as they show
high tolerance and reasonally good performance under coal-smoke pollution. It is
hoped that planting this broad leaved species particularly Ficus spp. and Cassia
fistula in large number at the pollution source may help to maintain the hygenic
atmosphere. Azadirachta indica a versatile tree of Indian origin is another perennial
form which could survive, clean and maintain greeneny in the pollution enriched
atmosphere.
100
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A P P E N D I X
The reagents for various biochemical determinations were prepared accord-
ing to the following procedure.
1. Reagents for Nitrogen estimation
1.1 Nessler's Reagent:-
10 g of potassium iodide was dissolved in 10 ml of distilled water. To this was
added a solution ofmercuric chloride (6 gin 100 ml of water) in smalHots and with
shaking till a slight permanent precipitate was formed. To this was addedSO ml of 9
N potassium hydroxide solution and then diluted to 200 ml with distilled water. The
solution was kept overnight and the clear solution decanted for use.
2. Reagents for Phosphorus estimation
2.1. Molybdate Reagent:-
6.25 g of ammonium molybdate was dissolved in 75 ml of ION H^SO^. To this
solution 175 ml of distilled water was added in order to get 250 ml of the above
reagent.
2.2. Amino naphthol sulphonic acid:-
0.5 gof 1, 2, 4, Amino-naphthol-sulphonic acid was dissolved in 195 ml Of
15% sodium bisulphite solution to which 5 ml of 20% sodium sulphite solution was
added. The above solution was stored in a dark coloured bottle.
3. Reagents for Protein estimation
3.1. Reagent A:-
2% Sodium carbonate was mixed with O.IN sodium hydroxide solution.
3.2. Reagent B:-
0.5% Copper sulphate (CuSO^.SH^O) was added to 1% potassium sodium
tartrate.
3.3. Reagent C (Alkaline Copper sulphate solution):-
It was prepared by mixing 50 ml of reagent 'A' and 1 ml of reagent 'B' prior
to use.
3.4. Reagent D (Folin-Ciocalteau reagent):-
100 g sodium tungstate (Na2Wo0^.2HjO) and 25 g sodium molybdate
(Na2Mo0^.2HjO) was dissolved in 700 ml distilled water in which 50 ml of 85%
phosphoric acid and 100 ml of concentrated hydrochloric acid was mixed. The flask
was connected with a reflux condenser and boiled gently on a heating mantlefor 10
h. At the end of the boiling period, 150 glithium sulphate, 50 ml distilled water and
3-4 drops of liquid bromine was added to this flask. The reflux condenser was
removed and the solution in the flask was boiled for 15 minutes in order to remove
excess bromine, cooled and diluted to 1 liter. The strength of this acidic solution (1
N) was tested by treating it with 1 N sodium hydroxide using phenolphthalein as an
indicator.
4. Reagent for Calcium and Sodium estimation
4.1 TAM solution (Tri acid mixture)
It is a mixture of three acids like nitric acid, sulphuric acid and per chloric acid
in the ratio of 10:5:4.
II