STUDIES ON LEAF BLIGHT OF BLACK
GRAM (Vigna mungo (L.) Hepper) OF
CHHATTISGARH PLAINS
M. Sc. (Ag.) Thesis
by
Manish Kumar Sahu
DEPARTMENT OF PLANT PATHOLOGY
COLLEGE OF AGRICULTURE, RAIPUR
FACULTY OF AGRICULTURE
INDIRA GANDHI KRISHI VISHWAVIDYALAYA,
RAIPUR (Chhattisgarh)
2018
STUDIES ON LEAF BLIGHT OF BLACK
GRAM (Vigna mungo (L.) Hepper) OF
CHHATTISGARH PLAINS
Thesis
Submitted to the
Indira Gandhi Krishi Vishwavidyalaya, Raipur
by
Manish Kumar Sahu
IN PARTIAL FULFILMENT OF THE REQUIREMENTS
FOR THE DEGREE OF
Master of Science in
Agriculture
(Plant Pathology)
U.E. IDNo.20161724933 ID No.120116177
JULY, 2018
ACKNOWLEDGEMENT
“Education plays vital role in personal and social development and
teacher plays a fundamental role in imparting education. Teachers have crucial
role in shaping young people not only to face the future with confidence but also
to build up it with aim and responsibility. There is no substitute for teacher pupil
relationship”. I take this golden opportunity to express my heartful humble and
deepest sense of gratitude to those who helped me to complete my research
possible. These words are small acknowledgement but never fully recompensed for
their great help and co-operation.
I start in the name of God-who has bestowed upon me all the physical
and mental attributes that I posses and skill to cut through and heal a fellow
human.
It is my privilege to study and conduct my research under Dr. N. Khare
Principal Scientist, Department of Plant Pathology, College of Agriculture,
Raipur (C.G.), Chairman of my advisory committee, who provided me the research
insight, illuminating and meticulous guidance, calm endurance, continuous and
unfailing encouragement, scholarly suggestions, unique supervision, constructive
criticisms, sympathetic attitude, plausible appreciation and sustained support
during the entire course of investigation and preparation of manuscript. I am
highly indebted to him for his invaluable painstaking efforts taken towards my
study while devoting his precious time. His scientific approach and generosity
without any reservation greatly helped me to work under his supervision,
knowledge and enthusiastic interest, which he provided me throughout my post
graduation and research investigation despite his busy schedule of work.
I emphatically and gratefully acknowledge extend my loyal and venerable
thanks to members of my Advisory Committee, Dr. A.S. Kotasthane, Professor
and Head (Department of Plant Pathology), Dr. P. L. Johanson, Senior Scientist,
(Department of Genetices and plant Breeding), Dr. N. Lakpale, Associate
Professor, Dr. R. K. Dantre, Professor, (Department of Plant Pathology) and Dr.
(Smt.) G. Chandrakar, Professor, (Department of Agril. Statistics and Social
Sciences, Language) for providing proper guidance, critical comments, valuable
suggestions and deligent support lead to timely completion of this work.
I am also highly obliged to Hon’ble Vice Chancellor, Dr. S. K. Patil, Dr.
S. S. Rao, Director Research Services, Dr. O. P. Kashyap, Dean, College of
Agriculture, Raipur, Dr.M.P. Thakur, Director of Instructions andDr.(major)
TABLE OF CONTENTS
Chapter Title Page ACKNOWLEDGEMENT
TABLE OF CONTENTS
LIST OF TABLES
LIST OF FIGURES
LIST OF PLATES
LIST OF ABBREVIATIONS
ABSTRACT
I INTRODUCTION
II REVIEW OF LITERATURE
2.1General
2.2 Collection, Isolation, Purification and
identification of pathogen Macrophomina
phaseolina
2.3 Screening of black gram entries for their
reaction to Macrophomina blight in-vivo
2.4 In-vitro evaluation of Pseudomonas
fluorescence on the radial mycelial growth
of Macrophomina phaseolina
2.5 In-vitro evaluation of fungicides on the
radialmycelial growth of Macrophomina
phaseolina
iii-vi
iv-v
vi
vii
viii
ix
x-xiii
1-4
5-21
5-6
7-8
8-10
11-15
16-21
III MATERIALS AND METHODS 22-27
3.1 Collection, Isolation, Purification and 22
identification of pathogen Macrophomina
Phaseolina
3.1.1 Cleaning and Sterilization of materials 22
3.1.2 Collection of disease sample 23
3.1.3 Isolation 23
3.1.4 Purification and maintenance of culture and 23
identification of pathogen
3.1.4.1 Media used 23
3.1.5 Identification of the fungus 24
3.2 Screening of black gram entries against 24
Macrophomina phaseolina
Chapter Title Page 3.3 In-vitro evaluation of Pseudomonas
fluorescence on the radial mycelial growth
of Macrophomina phaseolina
3.4 In vitro evaluation of fungicide on the
radialmycelial growth of Macrophomina
phaseolina
IV RESULTS AND DISCUSSION
4.1 Collection of disease sample
4.1.1 Isolation
4.1.2 Identification of the fungus 4.2 Screening of black gram entries for their reaction
to Macrophomina blight 4.3 In-vitro evaluation of Pseudomonas fluorescence
on the radial mycelial growth of Macrophomina phaseolina
4.4 In vitro evaluation of fungicides on the mycelialgrowth of Macrophomina phaseolina
V SUMMARY ANDCONCLUSION
REFERENCES
RESUME
25
26-27
28-47
28
29
29-34
35-37
38-42
43-47
48-49
50-58
59
LIST OF TABLES
Table No. Title Page No.
3.1 Categorization of entries on the basis of per cent disease 24
incidence and 1-5 scale (IIPR, Kanpur, 1996)
3.2 List and Concentration of fungicides used in in vitro 27
Evaluation
4.1 Pathogen isolated from infected leaf and stem samples of 28
different entries of black gram
4.2 Screening of blackgram entries for their reaction to 36
Macrophomina blight
4.3 Efficacy of differentPseudomonas isolateson the 39
growth of M. phaseolina in vitro
4.4 Effect of different fungicides on growth of 44
Macrophomina phaseolina
LIST OF FIGURES
Figure No. Title Page No.
4.4 In-vitro evaluation of Pseudomonas fluorescence on 41
the radial mycelial growth of Macrophomina phaseolina
4.5 Per cent reduction in mycelial growth over control 42
4.6 Effect of different fungicides on growth of 46
Macrophomina phaseolina in vitro
4.7 Per cent reduction in mycelial growth over control 47
LIST OF PLATES
Plates Title Page
4.1 Disease symptoms in field 31
4.2 Symptoms used for isolation of pathogen 32
4.3 Pure culture of Macrophomina phaseolina on PDA 33
4.4 Pure culture of Macrophomina phaseolina on water agar 33
4.2 Sclerotia of Macrophomina phaseolina 34
4.3 Screening of black gram entries 37
4.4 Effect of different Pseudomonas fluorescenceisolates on 40
growth of Macrophomina phaseolina in vitro
4.5 Effect of different fungicides on growth of Macrophomina 44
phaseolina in vitro
LIST OFABBREVIATIONS
Abbreviations Description
% : Per cent
0C : Degree Celsius
Cm : Centimetres
Fig. : Figure
Gm : Gram
ha-1 : Per hectare
i.e. : That is
Kg : Kilogram
m-2 : Per Meter square
Mm : Milometer
S. No. : Serial number
Viz : That is to say / in other words
q ha-1 : quintal per hectare
CD : critical difference
et al. : et alia (and others)
PDI : Percent disease Incidence
ppm : Part per million
LSI : Location Severity Index
SEm± : Standard error of means
Out of Forty entries screened, none were resistant or moderately resistant.
Only 4 entries i.e. KPU 17- 1, KPU 17- 2, KPU 17- 3 and KPU 17- 4 were
moderately susceptible. Rests of the varieties were susceptible and highly
susceptible against Macrophomina phaseolina.
Ten different isolates of Pseudomonas fluorescence were tested for their
antagonistic potential against Macrophomina phaseolina, the result revealed that
among all isolates isolate 1 exhibited in vitro maximum antagonistic potential and
significantly reduced mycelia growth (36mm) and percent inhibition (60%) of M.
phaseolina, followed by P3.
All the fungicides except propineb significantly reduced the mycelia
growth of Macrophomina phaseolina. Carboxine+Thiram, Hexaconazole+Zineb
and Tricyclazole completelyinhibited the mycelial growth. Thiram and
Carbendazim were found to be least and Propineb is not effective in reducing the
growth of the test fungus.
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CHAPTER - I
INTRODUCTION
Pulses in India have long been considered as the poor man’s only source of
dietary protein. Besides being a rich source of protein, they maintain soil fertility
through biological nitrogen fixation in soil and thus play a vital role in sustainable
agriculture (Kannaiyan, 1999). In India, pulses have been cultivated since time
under rainfed situations which is characterized by poor soil fertility and moisture
stress. These crops are energy rich but cultivated largely under energy starving
situations. Unlike in cereals, varietal breakthrough in pulses has not been taken
place. Pulses occupies 25.25 m ha area and contributes 16.46 m tonnes production
with an average productivity of 652 kg ha-1 (Anonymous, 2015-16a). During the
last four decades, the total area under pulses remained virtually stagnant (22 to 24
million ha) with almost stable production (12 to 14 million tonnes), even though
the population has been increased. As a result, per capita availability of pulses has
been declined from 64 g per day in 1951-56 to less than 40 g per day as against
WHO’s recommendation of 80 g per day (Asthana and Chaturvedi, 1999). This
situation led to the severe shortage of pulses in India, which has aggravated the
problem of malnutrition in large section of vegetarian population of our country.
Blackgram is one of the important kharif pulse crop grown throughout the India,
next to green gram. It is consumed in form of ‘dal’ (whole or split, husked or
unhusked) or parched. It is chief constituent of ‘papad’ and also of ‘bari’ (spiced
balls) which make a delicious curry. Urdbean differs from other pulses in its
peculiarity of attaining when mixed with water a somewhat mucilaginous pasty
character, giving additional body to the mass. In the south, the husked dal is
ground into a fine paste and allowed to ferment and is mixed with equal quantity of
rice flour to make ‘dosa’ and ‘idli’. It is also fried to serve as savoury dish. Urd dal
is also used in the preparation of ‘halwa’ and ‘imarti’.
Blackgram (Vigna mungo. L.) also known as urdbean or mashbean belongs
to popular plant family Papillionaceae and is among the most important pulse
crops of the world. It has been cultivated all over the world, mainly in south
western Asia, Egypt, Europe, India, Pakistan, Nepal and china. It has great value as
food, fodder and green manure. In addition to improving the soil fertility it is a
cheap source of protein for direct human consumption. The economic product of
black gram is seed grain, which is a good source of dietary protein. Urdbean
contains approximately 25-28% protein, 1.0-1.5% oil, 3.5 - 4.5% fibre, 4.5-5.5%
ash and 62-65% carbohydrates on dry weight basis. High values of lysine make
urdbean an excellent complement to rice in terms of balanced human nutrition.
Being short duration and photo, thermo insensitive, black gram is considered as an
excellentfor crop intensification and diversification.
Urdbean is one of the most important pulse crops of India cultivated over a
wide range of agro-climatic situations. Urdbean is mainly grown in tropical and
sub-tropical climate. Urdbean is a native of India and originated from wild plant
i.e. Phaseolus sublobatus. It occupies sizable area in India, Bangladesh, Pakistan,
Myanmar, Sri Lanka, and West indies. It is grown all over country in kharif and
summer seasons also, while in south India, it is raised mainly in rabi season.The
major urdbean growing states of the country are Maharashtra, Andhra Pradesh,
Madhya Pradesh, Uttar Pradesh, Tamilnadu, Karnataka and Rajasthan.
Development of short duration, photo, thermo-insensitive and disease resistant
varieties has led to its cultivation as a sole or intercrop during spring season in
North India and as a sole / relay crop during rabi season in the rice fallows of the
coastal peninsula. It is occupying an area of 3.06 million ha and total production of
1.70 million tonnes with an average productivity of 555 kg ha-1in India
(Anonymous 2014-15.c). In Chhattisgarh it occupies an area of 154.53 thousand ha
and total production of 128.00 thousand tonnes with average productivity 470 kg
ha-1 (Anonymous, 2014 – 15.c).
Throughout the India, the black gram is used for different purposes. The
major portion is utilized in making dal, curries, soup, sweets and snacks. With
sprouting there is an increase in the thiamine, niacin and ascorbic acid, thus
mungbean sprouts are increasingly becoming popular in certain vegetarian diets.
Moreover, its food values lie in high and easily digestible protein. Amino acid
analysis indicates that it is an excellent complement to rice for balanced human
nutrition.
The major fungal diseases which infect the crop are leaf blight
[Macrophomina phaseolina (Tassi) Goid], powdery mildew (Erysiphe polygoni
DC), web blight(Thanatephorus cucumeris (Fr.) Donk (=Rhizoctonia solani
Kuhn), Cercospora leaf spots (Cercospora canescens Ellis and Martin, C. cruenta
Sacc., C. dolichi Ellis and Everlast, C. kikuchi Matsumoto & Tomoyasu and
Anthracnose (Colletotrichumdematium and C. lindemuthianum (Philip et al., 1969,
Dwivedi and Saksena, 1974.,and Grewal,1988).
Macrophomina phaseolina (Tassi) Goid is one of the most damaging seed
andsoil borne pathogen, infecting about 500 plant species in more than 100
families throughout the world [(Kunwar et al, 1986, Mihail and Taylor 1995)].
Under favorable conditions the fungus causes many diseases like leaf blight,
damping off, seedling blight, collar rot, stem rot, charcoal rot and root rot in
various economically important crops. Mung bean was observed severely affected
by leaf blight caused by Macrophomina phaseolina (Tassi.) Goid. In Kharif as well
as during summer season.It was first reported from Jabalpur (M.P.) India (Philip et
al., 1969).
The pathogen attacks on all parts of plant i.e. root, stem, branches, petioles,
leaves, pods and seeds. Moreover, seed infection of Rhizoctonia bataticola (M.
phaseolina) ranges from 2.2-15.7% which causes 10.8% losses in grain yield and
12.3% inprotein content of seed in black gram (Kaushik et al.1987). The infected
seeds act as an important source of primary inoculum for new areas. Soil and seed
borne nature of the disease possesses problems for an effective disease
management. Therefore, an attempt has been made to integrate management of leaf
blight disease on black gram incited by Macrophomina phaseolina (Tassi.) Goid
which have become a serious problem in hampering the production of the black
gram in all growing areas of India.
Considering the severity of the disease in Chhattisgarh state and crop losses in
the farmer's field the present investigations entitled, “STUDIES ON LEAF
BLIGHT OF BLACK GRAM (Vignamungo (L.) Hepper) OF
CHHATTISGARH PLAIN”were undertaken with following objectives:
Objectives of investigation:
1. Collection, Isolation, Purification and identification of pathogenMacrophomina
phaseolina
2. Screening of black gram entries for their reaction to Macrophomina blight in-vivo
3. In-vitro evaluation of Pseudomonas fluorescence on the radial mycelial growth of
Macrophomina phaseolina
4. In-vitro evaluation of fungicides on the radial mycelial growth of Macrophomina
phaseolina
CHAPTER - II
REVIEW OF LITERATURE
In this chapter the literature on Macrophomina phaseolina has be reviewed
and cited below under different heading. Macrophomina phaseolina
(Rhizoctoniabataticola) is widely distributed throughout the tropical part of the
world. It is highlydestructive in nature and brings about severe losses in economic
crop plants.
2.1 General
Macrophomina phaseolina (Tassi) Goid(Tiarosporella phaseolina (Tassi)
Van der Aa) is a soil borne plant pathogenic fungus. It belongs to the anamorphic
Ascomycetes and is characterized by the production of both pycnidia and sclerotia
in host tissues and culture media. The pycnidial state was initially named
Macrophoma phaseolina by Tassi in 1901 and Macrophoma phaseoli by Maublanc
in 1905. In 1927, Ashby maintained the name Macrophomina phaseoli, while
Goidanich (1947) proposed Macrophomina phaseolina. Tiarosporella phaseolina
(Tassi) Van der Aa was used in 1981 by Van der Aa to designate the species. The
sclerotial state was described for the first time by Halsted as Rhizoctonia bataticola
(Taub.) Butler on Ipomoea batatas in 1890. Singh (1981) has given a brief review
on the origin and interrelationship of Vigna mungo and V. radiata. The symptoms,
mode of transmission, and host range ofimportant diseases, namely mungbean
yellow mosaic virus, leaf crinkle virus, leaf curl virus, mosaic mottle virus, and
diseases caused by Cercospora canescens, Erysiphepolygoni, Rhizoctonia
bataticola [Macrophomina phaseolina], R. solani, Xanthomonas phaseoli and
Pseudomonas phaseolicola are given. Screening forresistance, sources of
resistance, including interspecific hybridization, and induced mutations as well as
the genetics of resistance are treated along with suggestions for future breeding
strategies, symptoms, hosts and transmission of some virus, fungal and bacterial
diseases of Vigna radiata and V. mungo are outlined and information on breeding
and the genetics of resistance is reviewed. Sharma and Singh (2000) reported
that24 per cent of mungbean (Vignaradiata) seed samples collected from 11
districts of Rajasthan in India, during 1996-97, showed 0.5-38 % infection with
Macrophomina phaseolina (Rhizoctoniabataticola).
Rauf (2000) detected 24 seed born fungi belonging to different genera
using blotter paper method, from 145 seed samples of major legume crops in
Pakistan. Among these Macrophomina phaseolina, Alternaria alternata,
Ascochyta sp., Colletotrichum sp., Fusarium sp. andMacrophomina phaseolina
were the mostfrequent and known as common pathogenic fungi.
Khan et al. (2008) identified sources of genetic resistance in mung bean
against charcoal rot caused by Macrophomina phaseolina, 29 germplasm
accessions were evaluated by paper towel technique under laboratory conditions.
Two genotypes (NCM 252-10 and 40536) were highly resistant, whereas 5
cultivars (40504, NCM 257-5, 40457, NCM 251-4 and 6368-64-72) were resistant
and 6 were moderately resistant. Three genotypes were tolerant whereas the rest of
the accessions were susceptible or highly susceptible. The paper towel technique
proved to be quick and efficient for identification of resistance in mung bean for
charcoal rot disease.
Singh et al. (2013) had undertaken a study to screen for percent incidence
of seed borne mycoflora of two mung bean varieties viz. HUM-4 and HUM-7
They were screened by standard blotter paper and agar plate methods. Agar plate
method was found to be suitable as even under lesser incubation there was higher
observed percent incidence of seed mycoflora.
Suradkar S. et al. (2015) tested Twenty-seven varieties/ germplasms of
Black gram grown at Pulse Research Station, N.A.U. Navsari in Kharif season,
2008. The reactions of varieties/ germplasms against the disease were recorded. In
which TPU-4 (0.95 %) recording minimum per cent disease index was resistant
while eight germplasms were rated as moderately resistant. GU-1 (52.00 %), UB-8
(55.00%), UB-15 (56.00 %), UB-16 (60.00 %), showed susceptible reaction while
the germplasms UB-19 (78.00%) and UB-22 (76.33 %) showed highly susceptible
reactions. Other varieties showed moderately susceptible reactions.
2.2 Collection, Isolation, Purification and identification of pathogen
Macrophomina phaseolina
Philip et al. (1969) reported that Rhizoctonia bataticola infects the root's
hypocotyl region and leaf of the moong and urd crop.
Saxena et al. (1970) reported that same strains of Macrophomina
phaseolina (Rhizoctonia bataticola) caused blight and dieback of black and green
gram. It is alsoreported to cause leaf spot and blight in pigeon pea in North-
India.Sakuja (1974) recorded infection of mung and urdbean causing leaf blight
disease.
Jain et al. (1973) studied isolates of R. bataticola obtained from urd plant
parts (root, stem, leaf, pod and seeds) and soil. The isolates from various plant
parts and soil showed differences in virulence. The soil isolate was most
pathogenic. The isolates differed in their growth pattern and sclerotial size. The
leaf isolate developed the largest sclerotia and the seed and soil isolates developed
the smallest sclerotia. When grown on different media the isolates varied in growth
pattern and growth rate. The soil isolate showed the least amount of growth in
almost all media.
Hooda and Grover (1982) reported that isolates of Macrophomina
phaseolina obtained from different plant species and plant parts of the same host
differed in their morphological and cultural characteristics. There was no
correlation between these characteristics and their pathogenicity on Vigna radiata.
Young inoculum (3-5 days old) was more infective than old (7-34days) and with
increase in inoculum density disease intensity also increased. Macrophomina
phaseolina causing leaf spot of Vignaradiata crop was reported in Queensland,
Australia. The inoculum sources wasconsidered to be microsclerotia of the fungus
in soil splashed on the leaves (Fuhlbohm et al., 1986).
Devi and Singh (1998) revealed that total of 56 Macrophomina phaseolina
isolates were obtained form black gram and green gram crops. The isolates were
categorized as highly virulent (MP-2 and MP-3) moderately virulent (MP-1, MP-4,
MP-6) and weakly virulent (MP-5) on the basis of disease incidence and intensity
on black gram and green gram.
Kale (1999) isolated Rhizoctonia bataticola from infected leaf samples by
using tissue isolation method and pathogenicity was proved on three weeks old
mungbean plants.
Ahmadi et al. (2010) revealed that Macrophomina phaseolina is one of the
most important plant pathogenic fungi species that causes charcoal rot in many
plants. In September of 2009, during a study of epidemics of this disease in
Soybean fields of Golestan province, symptoms of charcoal rot on a number of
weeds was observed and after isolation and purification, the causal fungus was
identified as Macrophomina phaseolina. The weeds species includes Stinging
nettle (Urtica dioica), Black LaceElderberry (Sambucus nigra), wild mustard
(Sinapis arvensis) and camelthorn (Alhagimaurorum). All of four hosts are
introduced as new hosts for this fungus in Iran.
Iqbal and Mukhtar (2014) studied the morphological and pathogenic
variability among the 65 isolates of Macrophomina phaseolina from different agro
ecological regions of Punjab and Khyber Pakhtunkhwa provinces of Pakistan.
Characters taken for study were radial growth, sclerotial size, weight and
pathogenicity. Sixteen isolates were rated as fast growing, 11 as slow growing, and
the rest of the isolates as medium growing. Nine isolates were classified as large
sized, 26 as small sized, and the remaining 30 as medium sized. Ten fungal isolates
appeared to be least virulent, whereas eight isolates of diverse origin proved to be
highly virulent against mungbean cultivars. The remaining isolates were regarded
as moderately virulent. No relationship was found among the morphological
characters and pathogenicity of the isolates.
2.3 Screeningof black gram entries for their reaction to Macrophomina blight
in-vivo
Deshkar et al. (1974) screened 163 varieties of mungbean for their
resistance to Rhizoctonia bataticola (Macrophomina phaseolina) and reported that
none of the variety evaluated was resistant or moderately resistant. Only one
variety 11160 (a) was moderately susceptible and rests were susceptible.
In field trials with 19 tolerant lines of Phaseolus aureus (Vigna radiata)
only two showed high resistance to Macrophomina phaseolina (Vidhyasekaran et
al., 1977).
Sivaprakasam et al. (1983) screened 20 Vigna radiata genotypes for
reaction to Macrophomina phaseolina under artificial conditions, PIMS2, PIMS3,
PIMS4 andBGG2 proved to be resistant.
Zote et al. (1983) screened mung cultivars and found none of the cultivar to
be completely free from Macrophomina blight, however ML-5, ML-26, ML-65
and ML-62 were found to be moderately susceptible to Macrophomina blight. J-
781 was highly susceptible.
Shirshikar et al. (1991) conducted field trials over 3 years with 30
Vignaradiata cultivars exposed to artificial infection by M. phaseolina, only BCG-
1 wasconsistently highly resistant, 4 others being moderately resistant. BCG-1 may
be used in a breeding programme to develop high yielding resistant lines.
Suresh et al. (1992) observed that a pure line selection from local
Vignaradiata cv Kaveripattinam was identified as a high yielder and designated as
DPI-703.It matures in 85-90 days, withstands drought during early growth has an
erect, compact habit and broad green leaves. Resistance was shown to
Macrophomina phaseolina and Erysiphe polygoni.
Burman et al. (1998) observed that Vigna radiata genotypes PDM-86-199
and K-851 were planted in a loamysand soil with a 10.4% moisture holding
capacity in Jodhpur, Rajasthan in India. Genotype K-851 was more susceptible to
Macrophomina phaseolina infection than PDM-86-199.
Dhutraj et al. (2005) screened eighteen urd bean genotypes, along with two
controls (pusa vishal and barkha), for their resistance to powdery mildew (Erysiphe
polygoni) and Macrophomina blight (Macrophomina phaseolina) during kharif
2004 in Badnapur, Maharashtra, India. All genotypes were resistant to both
powdery mildew and Macrophomina blight. Other resistant sources can be utilized
in breeding programmes for developing disease resistance in urd bean.
Choudhary et al. (2010) conducted an experiment of screening during 2006
and 2007 to evaluate 25 greengram [Vigna radiata (L.) Wilczek] genotypes for
resistance to root rot caused by Macrophomina phaseolina (Tassi.) Goid. Complete
resistance to root rot could not be found, however, 'MSJ 118' genotype exhibited
highest suppression of dry root rot, followed by the genotype 'KM 4-59' and
appeared as moderately resistant genotypes.
Deepthi et al. (2014) revealed that Charcoal rot is one of the most
destructive disease which causes heavy loss in sesame. Macrophomina phaseolina
was identified as pathogen of charcoal rot disease on the basis of morphological
studies and pathogenicity. Among the evaluated sesame cultivars, none of the
cultivars showed complete resistance against M. phaseolina. Only one entry
PKDS-91 was found as moderately resistant to charcoal rot. Three entries (OSC-
366-I, SSD-2-I and OSC-79) were recorded as moderately susceptible.
Tak et al. (2015) screened Mungbean varieties for associated mycoflora
and its management. Three mungbean cultivars SML 668, ML 818 and PAU 911
showed association of mainly six fungi viz. Macrophomina phaseolina (52.66%),
Aspergillusflavus (39.32%), Aspergillus niger (15.33%), Aspergillus versicolor
(9.11%), Aspergillus clavatus (8.89%) and Penicillium sp. (6.00%). The maximum
mycofloraincidence was observed in mungbean cv. SML 668, where highest
incidence of M.phaseolina (38.44%) and Aspergillus flavus (28.44) was observed.
Suradkar S. et al. (2015) tested Twenty-seven varieties/genotypes of
Blackgram grown at Pulse Research Station, N.A.U. Navsari in Kharif season,
2008. The reactions of varieties/ germplasms against the disease were recorded. In
which TPU-4 (0.95 %) recording minimum per cent disease index was resistant
while eight germplasms were rated as moderately resistant. GU-1 (52.00 %), UB-8
(55.00%), UB-15 (56.00 %), UB-16 (60.00 %), showed susceptible reaction while
the germplasms UB-19 (78.00%) and UB-22 (76.33 %) showed highly susceptible
reactions. Other varieties showed moderately susceptible reactions.
2.4 In-vitro evaluation of Pseudomonas fluorescence on the radial mycelial
growth of Macrophomina phaseolina
Pande and Chaube (2003) reported that In vitro antibiosis of 6 isolates of
P.fluorescence resulted in reduction of mycelial growth of R. solani and the
inhibitionzone ranged from 1.3 to 22.5 mm in different isolates on Kings-B
medium.
Ahmadzadeh et al. (2006) evaluated biological control activity of 47
Pseudomonas fluorescence spp., against certain soil-borne phytopathogenic fungi
such as, M. phaseolina, R. solani, P. nicotianae var. parasitica, Pythium sp. and
Fusarium sp. Theresults indicated that 66%, 40.42%, 63.82%, 48.94% and 27.65%
of strains revealed antagonistic activity against R. solani, M. phaseolina, Pythium
sp., P. nicotianae and Fusarium sp., respectively.
Dhoke and Kurundkar(2006) tested the efficacy of isolates of P.
fluorescence against M.phaseolila on potato dextrose agar medium by dual culture
technique. In generalmycelia growth of bacterial isolates decreased and formed
zone of inhibition indicating their antagonistic nature. The most promising isolates
found were FP1 and FP2.
Rini and Sulochana (2007) isolated twenty-six isolates of Trichoderma spp.
and 56 isolates of Pseudomonas fluorescence from Kerala were evaluated for their
antagonistic activity against R. solani under in vitro conditions. Different isolates
showed varying degrees of antagonism. The two most antagonistic isolates against
R.solani were T. pseudokoningii TR17 and T. harzianum TR20 of the
Pseudomonas fluorescence, P. fluorescence isolates P28 and P51 showed the
greatest inhibition (26.6%) and (27.5%) against R. solani.
Goud and Muralikrishnan (2009) tested antifungal activity of P.
fluorescence against P. oryzae, P. ultimim and M. phaseolina. All three pathogenic
fungi were inhibited by P. fluorescence with inhibitory activities ranging from
50% to 80%.
Fifteen rhizobacterial Pseudomonas fluorescence isolates obtained from
rice in the region of Andhra Pradesh, India. In all 10 strains of Pseudomonas
fluorescence were selected based on preliminary screening of all these isolates for
antifungal activity against rice fungal pathogens (P. oryzae and R. solani) inhibited
the growth of rice fungal pathogens in Fe deficient King‟s B medium varied from
(3 to 58% inhibition). Among these Pf 003 strain completely inhibited the mycelial
growth of two rice pathogens both in presence and absence of FeCl3 which
indicated the siderophore mediation along with antifungal metabolites. (Battu and
Reddy, 2009).
Afsharmanesh et al. (2010) reported that Pseudomonas fluorescence able to
produce secondary antifungal metabolites can inhibit soil-borne plant pathogens.
For this reason, the antagonistic activity of P. fluorescence UTPF5 against R.
solani AG-4 was assessed in bean under in-vivo and in-vitro conditions. Production
of some secondary and nonvolatile metabolites on growth of the fungus were
observed in UTPF5 and their impact on mycelial growth of R. solani was also
studied. The results showed that UTPF5 could inhibit the growth of R. solani both
in-vitro and in-vivo, and suppress the disease by 33.34% and 14.29% by soil
drenching and seed treatment, respectively. Production of HCN, siderophore and
protease and involvement of siderophore, volatile and nonvolatile metabolites on
growth of the fungus were observed in UTPF5.
Dev and Dawande (2010) evaluated the antagonistic property of
Trichodermaspp. and P. fluorescence against R. solani and found that the
mycolytic enzymesproduced by the antagonists suppressed the growth of R. solani.
Two hundred isolates from cotton rhizosphere isolated by Fallahzadeh and
Ahmadzadeh (2010), out of which 39% pertained to Pseudomonas fluorescence.
Dual culture assays were conducted against R. solani AG4 for these isolates to
evaluate the ability of antibiotic production. In greenhouse studies, all of the
isolates significantly suppressed the disease on plant.
Pan and Josh (2010) studied that the inhibition in growth of Trichoderma
was 8.59% when Trichoderma was used first and 59.44% when P. fluorescence
was used first in a dual culture test. Highest growth inhibition (59.44%) of
Trichoderma (Th3) was recorded when P. fluorescence was first inoculated in
King's medium B agar (KMB) 24h prior to inoculation of T. harzianum.
Malhotra et al. (2011) evaluated thirteen bio-control fungi and 4 bacterial
strains against R. solani using dual culture technique. The results showed that
among 12 the fungal species Gliocladium virens and T. harzianum (T8) were the
most effective isolates and inhibited R. solani mycelial growth by 74.82% and
73.33% respectively. Among the bacterial strains maximum growth inhibition was
caused by P. fluorescence P.f.1 (73.33%) followed by P. fluorescence P.f.2
(62.22%).
Rajeswari and Kannabiran (2011) reported that T. viride, T. harzianum, and
P. fluorescence were evaluated for their antagonistic activity against F. oxysporum
in-vitro. Among them, highest percent inhibition of conidial germination was
brought out by T. viride (89.4%) followed by T. harzianum (85.7%) and P.
fluorescence (83.15%) and inhibition of radial mycelial growth were (86.6%),
(84.0%), (60.0%) respectively.
Singh (2011) selected antagonists isolates i. eTrichoderma (Th38 and
Tv34), Pseudomonas fluorescence (Pf5 and Pf7), Bacillus subtilis (Bs5), non-
pathogenic Fusarium (Fo52) and Epicoccum purpurescens (Ep5) as potential bio-
control agents. The selected antagonists are highly effective against R. solani,
Sclerotinia sclerotiorum, M. phaseolina and F. oxysporum causing diseases in
vegetable crops.
Gull and Hafeez (2012) examined 28 Pseudomonas bacterial strains,
among the 28 strains tested, 14 were found to be siderophore producers. These
strains were evaluated for their bio-control potential against R. solani using various
dual culture assays. The role of siderophores in the inhibition of R. solani was
confirmed by iron chloride (FeCl3) experiment. Data demonstrated that bacterial
strain Mst 8.2 produces more than one antifungal agent but the siderophore
production is the key mechanism involved in the antagonism. Bacterial strains MS-
3y, Mst 8.2 and Mst 7.4 were the most effective with more than 70% disease
reduction in wheat.
Kapoor et al. (2012) determined the efficacy of different microbes for their
antagonistic ability in-vitro against F. oxysporum f. sp pisi by dual culture method.
The antagonists inhibited the mycelial growth of F. oxysporum f. sp. Pisi.,
T. viride (51.55), P. fluorescence-1 (49.62) and P. fluorescence -2 (50.08) showed
maximum inhibitionantagonistic activity in vitro. In-vivo T. viride and P.
fluorescence resulted in maximum reduction of seed rot root and wilt (F.
oxysporum) of pea.
Ten isolates of bacteria, designated as PGB1, PGB2, PGB3, PGB4, PGB5,
PGT1, PGT2, PGT3, PGG1 and PGG2, were successfully isolated and
characterized by Manivannan et al. (2012). Subsequently, to investigate the PGPR
isolates for their antagonistic activity against phytopathogenic fungi such as F.
oxysporum, R. solani and Sclerotium rolfsii. Furthermore, most of the PGPR
isolates shown antifungal activityagainst F. oxysporum, R.solani, and Sclerotium
rolfsii.
Adhikari et al. (2013) evaluated seventy isolates, antagonistic twenty-one
representing bio-vars of P. fluorescence (bio-vars I, II, III, and V) were collected
from the rhizosphere of okra, chilli, ground nut, brinjal, cabbage and tomato from
different agro-ecological regions of West Bengal and were subjected to evaluate
for their antifungal activity under in vitro condition against R. solani, the most
important soil-borne plant pathogen. Two isolates, PF-8 and PF-7 effectively
inhibited the mycelial growth of R. solani (72.05 and 68.25%, respectively) in dual
culture method.
Arumugam et al. (2013) isolated P. fluorescence and T. viride from the
rhizosphere of rice fields by the serial dilution method and tested against rice
pathogens for R. solani, Helminthosporium oryzae and S. oryzae in dual culture
method. The test result revealed that P. fluorescence actively inhibited,
Heminthosporium oryzae 75.6%, R. solani 47.8%, S. oryzae 68.9%.
Ravindran and Shaike (2013) isolated R. solani from naturally infected
vanilla plants and attempted to minimize the damage caused by the pathogen using
bio-control agents T. harzianum and P. fluorescence isolated from soil. The
combined inoculation of T. harzianum with P. fluorescence treatment showed
maximum disease suppressionfollowed by the single inoculation of P.
fluorescence, T. harzianum, P. putida and T.virens respectively in decreasing
order.
Saravanan et al. (2013) focused on the antagonistic potential of
Pseudomonas fluorescence in vitro and its inoculation effect on growth
performance of Lycopersicon esculentum in R. solani infested soil. Isolates Pf5 and
Pf6 wereantagonistic against 14 bacterial species, and two pathogenic fungi (F.
oxysporum and R. solani).
Thirty isolates were screened for PGP attributes and isolates showing PGP
properties were further screened for in vitro antagonism against soil borne phyto-
pathogens via, R. solani, Sclerotium rolfsii and Fusarium spp. Results revealed that
50% (15 out of 30 isolates) reacted positively for one or more PGP properties. A
high prevalence of antagonists was found against the three fungal pathogens.
Majority of the bacterial isolates (33%) displayed antagonism through the
production of siderophores or HCN. (Sarvani and Reddy, 2013)
Akter et al. (2014) isolated 325 bacteria and 14 isolates were found to be
antagonistic against the pathogen, and in the dual culture test selected bacterial
isolates KMB25, TMB33, PMB38, UMB20 and BMB42 showed 68.44%, 60.89%,
60.22%, 50.00% and 48.22% fungal growth inhibition, respectively. These
bacterial isolates were identified as Pseudomonas fluorescence by morphological
and biochemical characterization.
Four bacterial strains of P. fluorescence were isolated from tomato field
soil by Mezeal (2014). The antagonistic microorganisms against the pathogens
were observed by dual culture technique. P. fluorescence5 isolate was found to
show 81.3% and 77.4% of growth inhibition against R. solani and F. oxysporum
respectively.
Maurya et al. (2014) found that total eight micro flora resembling
P.fluorescence were isolated and three isolates were confirmed as P. fluorescence
(strainP.f.01, strain P.f.05 and strain P.f.07). P. fluorescence strains P.f 07 were
found most effective with the highest antagonistic activity against three fungal
pathogen and show maximum inhibition of mycelial growth of R. solani (68.23%),
F. moniliforme (65.45%) and Alternaria alternate (48.13%)
Solanki et al. (2014) were screened 220 bacteria isolated from tomato
rhizosphere for in vitro antagonistic activity against R. solani AG-4. Five potent
antagonistic strains viz., Pseudomonas spp. (M10A and MB65), P. aeruginosa
(MPF14 and MB123) and P. fluorescence (MPF47) were identified.
2.5 In-vitro evaluation of fungicides on the radial mycelial growth of
Macrophomina phaseolina
Jhooty and Bains (1972) evaluated systemic and non-systemic
fungitoxicants against Rhizoctonia solani in-vitro. They found that Dithane M-45
at 10 µg/ml and 44 µg/ml inhibited the growth of fungus completely and 50%
respectively.
Kapoor and Chohan (1974) concluded that Thiram when mixed with Agar
medium and incubated at 30°C inhibited growth of Macrophomina phaseolina
completely.
Goel and Mehrotra (1981) tested nine fungicides in-vitro against
Rhizoctoniabataticola (Macrophomina phaseolina) of gram and found that bavistin
could inhibitthe fungal growth completely. Among 39 fungicides tested in the
laboratory studies carbendazim, benomyl, guazatine, dichlozoline and iprodione
were highly toxic to mycelial growth of Rhizoctonia bataticola (Macrophomina
phaseolina) in Czapek's medium (Hooda and Grover, 1983).
Martin et al. (1984) tested chlorothalonil at 0.1% against 16 isolates of
Rhizocatonia solani and reported that they were sensitive to chlorothalonil. Among
the non-systemic fungicides tested, mancozeb followed by copper oxychloride
were most effective in inhibiting the growth of Rhizoctonia bataticola
(Macrophomina phaseolina) (Patil and Wangikar, 1984).
Gautam and Narain (1986) tested efficacy of seven fungicides against
Macrophomina phaseolina causing blight of cowpea and found that bavistin,
thiramand topsin-M inhibited growth of fungus completely.
Basu et al. (1988) tested fourteen fungicides and found that Dithane-M-45
increased inhibition of fungus with increase in concentration.
Giri and Peshney (1993) tested a fungicides in-vitro and found that
carbendazim (0.1%) and mancozeb (0.25%) inhibited the spore germination and
mycelial growth of Colletotrichum graminicola and Rhizoctonia bataticola
(Macrophomina phaseolina) causal agent of leaf spot of mungbean.
Carbendazim and Thiophanate methyl at 0.2% were most effective growth
inhibitors of Macrophomina phaseolina causing seedling mortality of black gram
(Devi and Singh, 1997).
Kanakamahalakshmi et al. (1998) determined the effect of root rot caused
by M. phaseolina at different stages of castor bean growth. In addition, the
economiclosses due to the disease and its in vitro management using Carbendazim,
Thiophanate-methyl, Captan and Thiram. All the growth yield parameters were
adversely affected by M. phaseolina. Incidence of root rot at early stages of crop
growth drastically reduced plant height, spike formation, yield and oil contents.
Thiophanate-methyl, Carbendazim, Thiram and Captan were equally and highly
effective at 1500 ppm inhibiting the growth of M. phaseolina completely. The
fungicides were effective against M. phaseolina in vitro using poisoned food
technique were further evaluated in the laboratory to test the efficacy on seed
germination and seedling vigour index. Both systemic and non-systemic fungicides
tested were effective in increasing seed germination, hypocotyl length and root
length thereby reducing abnormal seedling production including browning and
blackening.
Lokesha and Benagi (2004) tested seven fungicides (Benomyl,
carbendazim, Carboxin, Thiophanate-methyl, Captan, Thiram and Mancozeb) at 3
different concentrations (250, 500 and 1000 ppm) for their efficacy against
Macrophomina phaseolina using the poison food technique in vitro. At 250 ppm,
benomyl recordedthe highest percentage of mycelial growth inhibition (98.14%),
followed by carbendazim (84.07%). At 500 ppm, both benomyl and carbendazim
fully inhibited the growth of the fungus (100%), followed by mancozeb (97.40%)
and thiram (97.03%). Benomyl, carbendazim, thiram and mancozeb showed 100%
growth inhibition at 1000 ppm. All fungicides were tested in pot culture
experiments conducted in 2002 in Raichur, Karnataka, India. Pigeon pea (ICP-
8863) seeds were treated with individual fungicides at 2 gkg-1
and sown (8 seeds
per pot) in earthen potscontaining sterilized soil mixed with 10% pathogen
inoculum. Maximum plant survival at 130 days after sowing was obtained with
both carbendazim and benomyl (both 86.70%), followed by mancozeb (82.02%).
Raut and Patil (2005) conducted a field experiment in Thane, Maharashtra,
India, during the 2002/ 03 rabi season on tomato cv. Pusa Ruby to evaluate the
efficacy of fungicides, botanicals and Trichoderma viride against Fusarium
oxysporum and Rhizoctonia bataticola [Macrophomina phaseolina]. The
treatments comprised 0.1%carbendazim, 0.1% Bordeaux mixture, 2.5 g T. viride
per plant, 0.1% Reviver, 0.1% Chetana and 0.1% garlic extract. Bordeaux mixture
gave the lowest disease incidence (22.22%) and highest tomato yield (17.12 tha-1
).
Khan and Khan (2006) determined the efficacy of several fungicides
against Macrophomina leaf spot (Macrophomina phaseolina) of mung beans in
vitro and inthe field (Uttar Pradesh, India). For the in vitro tests, treatments
comprised: carbendazim, Topsin M [thiophanate-methyl], benomyl, captafol,
mancozeb and thiram all at either 0.10 or 0.25% concentration and an untreated
control. For the field tests, treatments comprised: 0.10% carbendazim + seed
treatment with thiram (ST; 3 g/kg); 0.10% Topsin M + ST; 0.10% benomyl + ST;
0.25% captafol + ST; 0.25% mancozeb + ST; and an untreated control.
Carbendazim, Topsin M and benomyl werehighly effective against the pathogen in
vitro and recorded 100% pathogen radial growth inhibition at both concentrations.
At 0.10 and 0.25% concentrations, thiram recorded 89.7 and 95.5% pathogen
radial growth inhibition, captafol recorded 85.1 and 93.8% growth inhibition and
mancozeb recorded 78.7 and 83.1% growth inhibition, respectively.
Konde et al. (2008) reported that Soybean (Glycine max (L.) Merill) is
important oil seed crop in India. Rhizoctonia bataticola (Pycnidial stage -
Macrophomina phaseolina) is the important soil borne pathogen causes
rootrot/charcoal rot disease in soybean. In vitro studies of fungicides and bioagents
efficacy against Rhizoctonia bataticola, revealed carbendazim+thiram (0.1+0.2%),
penconazole (0.1%), thiophanate-M (0.1%) completely inhibited (100%) the
growth of pathogen. Among bioagents, Trichoderma viride inhibited the growth of
Rhizoctoniabataticola to the extent of 96.39 per cent in pot culture studies,
maximum germination(84.45%) and per cent disease reduction (52.39%) was
recorded in the seed treatment with thiram+carbendazim (2±1 g kg-1
).
Khalikar et al. (2011) observed that chemical control is one of the measures
to manage the disease and avoid the losses. The evaluation study was therefore
conducted in vitro. Seven fungicides were tested against the pathogen i.e.
Macrophomina phaseolina in vitro. The highest inhibition (100%) of M.
phaseolina was observed due to carbendazim (500 ppm), chlorothalonil (500 ppm),
hexaconazole (500 ppm) and captan (2500 ppm) followed by mancozeb (2500
ppm) (94.39%) and benomyl (1000 ppm) 93.4% and rest of the treatments
significantly inhibited colony growth over control. The significantly highest
inhibition (100%) of scleortial production was recorded due to carbendazim (500
ppm), chlorothalonil (800 ppm), hexaconazol (500 ppm) and captan (2500 ppm)
followed by mancozeb (2500 ppm) 96.59% and benomyl (1000) 96.59%.
Dhingani et al. (2012) tested eleven fungicides of four different categories
viz., six systemic, two non-systemic and three mixed formulations and six
herbicides at their three different concentrations in vitro by poisoned food
technique for evaluatintheir efficacy against M. phaseolina causing root rot of
chickpea. Among all concentrations tested, the higher concentrations of each of
fungicides and herbicides produced maximum growth inhibition of the pathogen.
From tested fungicides, carbendazim (Bavistin 50 WP), tricyclazole (Beam 75
WP), propiconazole (Tilt 25 EC), Quintal 50%WP (carbendazim 25%+iprodine
27%) and Sixer 75WP (carbendazim 12%+mancozeb 63%) at all three
concentrations completely inhibited growth of the pathogen and proved to be
highly toxic to the pathogen. Among all herbicides tested, pendimethalin (Stomp
30 EC), oxyflourfan (Galagan 23.5 EC) and alachlor (Laso 50 EC) were proved to
be effective in inhibiting the mycelial growth of the pathogen.
De R. K. (2014) tested new fungicides for management of stem rot of jute
(Corchorus olitorius L. and C. capsularis L.), caused by Macrophomina
phaseolina, revealed that pre-sowing seed treatment followed by foliar spraying of
tebuconazole one month after sowing resulted in lowest stem rot incidence of 25%
and it was statistically at par with carbendazim (28.5%) and hexaconazole (28.6%)
compared to 45.2% in check. These were best fungicides against jute stem rot
pathogen as also observed in in vitro tests. These were followed by Tricyclazole,
Copper oxychloride and Mancozeb, respectively, with 33.4, 33.5 and 35.9% of
stem rot incidence. Among the fungicides tested, Thiophanate methyl was least
effective against stem rot of jute with 38.7% disease. The progress of stem rot was
slowest in Tebuconazole and Carbendazim in all the dates of crop growth from 30
to 90 days after sowing. Chemicals and fungicides causing complete inhibition of
M. phaseolina under in vitro tests were Propiconazole 25% EC (10 µg/ml),
turmeric oil (10 µg/ml), Carbendazim 50 WP (25 µg/ml), Copper oxychloride 50
WP (50 µg/ml), Tebuconazole 25.9% EC (50 µg/ml), Hexaconazole 5% EC (100
µg/ml), curcumin mixture (100 µg/ml) and tricyclazole 75% WP (10000 µg/ml).
Deepthi et al. (2014) revealed that out of different fungicides evaluated
under in vitro conditions against M. phaseolina; vitavax power and penflufen gave
100%inhibition at 500 ppm, while tricyclazole gave 100% inhibition at 1000 ppm
and otherfungicides were less effective. The field evaluation of different fungicides
indicated that vitavax power gave highest seed germination and less pre and post
emergence mortality, and yield loss, whereas the sesame seeds treated with vitavax
power alongwith one foliar application of carbendazim was found most effective
for enhancing seed germination and reducing pre, post emergence mortality and
losses in yield of sesame.
Pawar et al. (2014) evaluated efficacy of the recommended pesticides viz.,
fungicides, and botanicals on the growth and sporulation of Rhizoctonia bataticola
causing (charcoal rot), of soybean under in vitro condition was. Using poisoned
foodtechnique, radial growth and visual observations were taken. Pyraclostrobin,
Picoxystrobin, Azcoxystrobin are highly effective in reducing myecial growth and
sclerotial formation of Rhizoctonia bataticola. Out of eight fungicides tested only
(carboxin, copper oxychloride, mancozeb, thiram, carbendazim) were effective in
reducing the growth of Rhizoctonia bataticola. Nilgiri, neem and ashoka were
moderately effective in reducing the mycelial growth and sclerotial formation of
Rhizoctonia bataticola.
Sangappa and Mallesh (2016) reported that Blackgram (Vigna mungo L.) is
an important pulse crop grown throughout India. A new disease of blackgram i.e.
aerial blight and dry root rot caused by Rhizoctonia bataticola is primarily a soil
inhabitant. Fungicides like contact, systemic and combi products were also tested
against the aforementioned pathogen. Among five contact fungicides captan,
propineb and zineb recorded cent per cent inhibition (100%) of mycelial growth at
all the concentrations (i.e., 0.1, 0.2 and 0.3% respectively). Among seven systemic
fungicides and combi fungicides tested, benomyl, carbendazim, hexaconazole,
thiophanate methyl and tridemefon showed 100 per cent mycelia inhibition and
also in carbendazim 12% + mancozeb 63%, cymoxanil 8% + mancozeb 64%,
captan70% + hexaconazole 5%, tricyclozole 18% + mancozeb 62% and mancozeb
(64%) + metalaxyl (4%) showed cent per cent (100%) inhibition at all the
concentrations (0.05, 0.10 and 0.2%), respectively.
CHAPTER- III
MATERIALS AND METHODS
The materials used in the present investigation and methods followed are given
below.
All the in-vitro studies on Macrophomina phaseolina (Tassi) Goid. of
Black gram, were conducted in the Department of Plant Pathology, Indira Gandhi
Krishi Vishwavidhyalaya, Raipur (C. G.). The field studies were conducted in the
Pulse Pathology Field, Block Number-23 of I.G.K.V., Research Farm of the
University. Glassware's were of Borosil Grade and chemicals of standard trade
markes (BDH, Qualigens and Merck, etc.) were used during the course of the
investigations. The following instruments were used in the present studies:
1. Autoclave for sterilization.
2. BOD incubator for incubation.
3. Hot air oven for glassware sterilization
4. Laminar flow for isolation.
5. Anamid electronic digital balance for weighing
3.1 Collection, Isolation, Purification and Identification of
pathogen Macrophomina phaseolina 3.1.1 Cleaning and Sterilization of materials
Prior to use glasswares were cleaned with detergent powder and finally
rinsed with tap water as per requirement of the experiment. The dried glassware’s
were sterilized in hot air oven at 180°C for two-to-three hours. The forceps and
other metallic instruments were sterilized by dipping them in alcohol and heating
over the flame before using them, sterilization of the media was done by
autoclaving at 1.02 kg/cm2 pressure for 20 minutes. The plastic plates were
sterilized with alcohol and air dried before use.
3.1.2 Collection of disease sample
The disease samples of Black gram having dark brown irregular lesions
were collected from the field of Pulse Pathology Research Farm, I.G.K.V., Raipur.
3.1.3 Isolation
The isolation of pathogen was made from the disease-infected leaf and stem
collected from the field of Pulse Pathology Research Farm. The usual tissue
isolation method was followed for the isolation of the fungus from leaves, infected
branches and stem. Infected leaf bits of black gram were first washed with tap
water and then with distilled water. The bits were then surface sterilized by dipping
in 0.1% Mercuric Chloride solution for a minute and again washed by giving three
successive changes of sterilized distilled water to remove the traces of Mercuric
Chlorides. The isolation work was carried out by using laminar airflow. The leaf
and stem bits were then placed on sterilized potato dextrose agar in Petri plates.
Plates were then incubated at room temperature (28±2°C). As soon as the growth
of fungus was observed in plates, small portion of mycelial growth was transferred
on potato dextrose agar slants. Number of slants were prepared for further
investigation.
3.1.4 Purification and maintenance of culture and identification of pathogen
3.1.4.1 Media used
Two per cent Water Agar and Potato Dextrose Agar was used for isolation,
purification, maintenance and morpho-cultural studies of isolates.
Water Agar
Agar agar : 20 gm
Distilled water : 1 liter
For preparing two percent water agar, 20 gm of agar-agar is added to one
literof water and mixed thoroughly. The medium was sterilized in an autoclave at
1.02 kg/cm2 (121 ºC) for 20 minutes.
Potato dextrose agar
Potato (peeled, washed and sliced) : 200g
Dextrose : 20g
Agar agar : 20g
Distilled water : 1000ml
For preparing PDA medium, 200g Potato washed, peeled off the skin and
sliced them into small pieces. The sliced potato was boiled in 500 ml water in a
vessel for 20 minutes. Simultaneously 20g of agar was mixed in 500 ml of water
and boiled for 30 minutes. The potato extracts was collected by filtering through
muslin cloth. Twenty gm of dextrose was added to the potato extract, mixed
thoroughly with potato-agar mixture and volume was made upto 1 liter with
distilled water. The pH of medium was checked by using pH metre, adjusted the
pH by adding 1 N HCl or 1 N NaOH as the case may be. The medium was
sterilized in an autoclave at 1.02 kg/cm2 (121 ºC) for 20 minutes.
King’s B medium,
A selective one (Kings et al., 1954) was used for the isolation and
multiplication of P. fluorescence. The composition of the medium was as
follows:
Ingredients Quantity
Protease peptone 20.0 gm
Dipotassium hydrogen phosphate 1.5 gm
Magnesium sulphate 1.5 gm
Glycerol 10.0 ml
Agar-Agar 20.0 gm
Distilled water 1000.0 ml
3.1.5 Identification of the fungus
Colony colour of Macrophomina phaseolina varies in culture from black to
brown or gray and becomes dark in color with age. Abundant aerial mycelium
isproduced in the culture plate with sclerotia imbedded within the hyphae
orengrossed in the agar or on the agar surface with smooth precincts. Hyphae are
septate, initially hyaline turning to a honey or black color. Numerous dark
brownsto black colored sclerotia can be seen on the reverse side of the culture
plate. Thevegetative mycelium is characterized by the formation of monilid or
barrel-shapedcells and the formation of septum near the branching of the
mycelium. Branchingoccurs at right angle to parent hyphae, but branching at acute
angles is also common (Dhingra and Sinclair, 1977).Isolates were given to name
on the basis of genus and species name of M.phaseolina i.e. MP.
The fungus was purified by repeated isolation from the culture plates. The
pathogen was identified on the basis of character of the mycelium and sclerotia.
The characters were compared with the standard description of Macrophomina
phaseolina from literature (Singh, 1998).
3.2 Screening of black gram entries against Macrophomina
phaseolina
Forty entries of black gram were sown with two replications. Each of the
mungbean entries were sown by dibbling at a distance of 10cm x 30cm. The
observation on incidence of disease was recorded at flowering and pod initiation
stage. Per cent incidence was recorded on the basis of visual observation. The
entries were categorized as follows.
Table 3.1 Categorization of entries on the basis of per cent disease incidence and 1-5 scale (IIPR, Kanpur, 1996)
S. No. Score PDI Category
1 1 0 Resistant
2 2 0.1-10.0 Moderately resistant
3 3 10.1-25.0 Moderately susceptible
4 4 25.1-50.0 Susceptible
5 5 Above 50.0 Highly susceptible
3.3 In-vitro evaluation ofPseudomonas fluorescence on the radial
mycelial growth of Macrophomina phaseolina
In-vitro evaluation of Pseudomonas fluorescence on the radial mycelial
growth of Macrophomina phaseolina was done by dual culture method to test
the efficacy as bio-control agent against M. phaseolina. Pseudomonas
fluorescence isolates were multiplied on King's B broth and incubated for 2 days
at 280C till the fluorescent pigment appeared in the broth. Equal volume of
sterilized potato dextrose agar (PDA) and King's B medium were mixed in
sterilized Perti plates. The edge of a glass funnel was sterilized by dipping in
alcohol followed by flaming. Broth containing young growing cell of
Pseudomonas fluorescence was dispensed in sterile Petri plate and picked at
the edge of the funnel by dipping. Care was taken to remove the excess
inoculum by gently shaking the funnel. Inoculation was done by gently
touching the edge of the funnel (containing Pseudomonas fluorescence) which
encircled the pre-inoculated with M. phaseolina on was placed in centre of the
plate. Control was also maintained to see the difference. Observation was taken
after three days of inoculation.
In vitro antagonistic potential of different isolates of Pseudomonas
fluorescence was assessed by calculating percentage inhibition growth of
pathogen (M. phaseolina) over control in presence of Pseudomonas fluorescence
using the formula of Vincent (1947):
C-T
Per cent inhibition = I = --------- X 100
C
Where,
I = Per cent inhibition
C = Radial growth of pathogen in control
T= Radial growth of pathogen in presence of Pseudomonas fluorescence
3.4 In vitro evaluation of fungicide on the radial mycelial growth of
Macrophomina phaseolina
Six fungicides were tested in vitro against M. phaseolina i.e. Thiram,
Carbendazim, Carboxin+Thiram, Tricyclazole, Propineb, Hexaconazole+Zineb.
Potato dextrose agar medium was sterilized in conical flask of 250 ml capacity.
Prior to pouring, fungicides Thiram(1%), Carbendazim (1%), Carboxin+Thiram
(2%), Tricyclazole (1%), Propineb(2%)and Hexaconazole+Zineb(1%) of active
ingredient were added to 100 ml of PDA seperately. The medium was then poured
in sterilized petriplates. Five mm discs of the test pathogen cut from the margin of
seven days old culture were placed centrally in each of the petriplate. The disc was
kept inverted to allow the contact of the fungus with the medium. The inoculated
petriplates without fungicides served as control. The inoculated plates were kept in
incubator at 28±2°C. Colony diameter of the pathogen was measured after 4, 6 and
8 days of inoculation with the help of scale and sclerotial formation was recorded
after 8 days of inoculation. Three replications of each treatment were maintained.
The Petri plates were incubated at 28±2 °C temperature till the complete coverage
in control plate. The per cent growth inhibition (PGI) of the pathogen was worked
out by using formula given by Vincent (1947).
C-T
Growth inhibition = I = --------- X 100 C
Where,
I= Percent inhibition
C = Mycelial growth in control plate
T = Mycelial growth in treatment plate
Table 3.2 List and Concentration of fungicides used in in vitro evaluation
S. No. Fungicides
Concentration in %
1. Thiram 2%
2. Propineb 2%
3. Carbendazim 1%
4. Tricyclazole 1%
5. Carboxin+Thiram 2%
6. Hexaconazole+Zineb 1%
7. Control -
CHAPTER-IV
RESULTS AND DISCUSSION
Present investigation on Macrophomina blight of blackgram was carried
out during2017-18 in laboratory and experimental field of Dept. of PlantPathology,
College of Agriculture Indira Gandhi Krishi Vishwavidyalaya, Raipur and the
results of the experiment are presented and discussed in the light of findings of
earlier researches in this chapter.
4.1 Collection of diseased sample
The Macrophomina blight disease samples of leaf and stem were collected
from thirteen different entries and subjected to isolation of the causal organism.
The details of the pathogen isolations were made from different entries and parts of
the plant are given in table 4.1.
Table 4.1: Pathogen isolated from infected leaf and stem samples of different
entries of Blackgram
S. No. Entries
Plant Parts
stem and leaf
No. of samples
Collected
M. phaseolina
Isolated
1. KPU 17- 6 Stem 2 Nil
Leaf 2 2
2. KPU 17- 8 Stem 2 Nil
Leaf 2 1
3. KPU 17- 14 Stem 2 1
Leaf 2 2
4. KPU 17- 18 Stem 2 Nil
Leaf 2 2
5. KPU 17-19 Stem 2 1
Leaf 2 2
6. KPU 17- 24 Stem 2 Nil
Leaf 2 1
7. KPU 17- 26 Stem 2 Nil
Leaf 2 2
8. KPU 17- 27 Stem 2 Nil
Leaf 2 2
9. KPU 17- 29 Stem 2 Nil
Leaf 2 1
10. KPU 17- 32 Stem 2 1 Leaf 2 2
11. KPU 17- 34 Stem 2 Nil
Leaf 2 2
12. KPU 17- 36 Stem 2 Nil
Leaf 2 2
13. KPU 17- 39 Stem 2 Nil
Leaf 2 1
Total no. of stem samples 26 3
Total no. of leaf samples 26 22
Two disease samples from stem and leaf of each variety were collected.
Macrophomina phaseolina could only be isolated from leaf samples of all the
varieties because the pathogen was observed to be more pre-dominant on leaf as
compared to the stem. Therefore, the pathogen isolated from leaf was used for
further investigation.
4.1.1 Isolation
Isolation of the fungus was done in Petri dishes using Potato Dextrose Agar
medium. The surface sterilized diseased bits yielded the fungus after 48 hours of
incubation at 28 ±2°C temperature. The uniform colonies originating from diseased
bits were separated, purified and were maintained on potato dextrose agar slants
for further use during entire course of investigation.
Purification
Purification of the fungus was done in Petri dishes using Water Agar
medium. The uniform colonies originating from diseased bits were separated, and
allowed to grow on Water Agar plates for single sclerotia isolation and purification
of pathogen and were maintained on potato dextrose agar slants for further use
during entire course of investigation.
4.1.2 Identification of the fungus
The pathogen was identified on the basis of character of the mycelium and
sclerotia. The mycelium was septate and hyaline, the sclerotia were brown to black
in colour, rounded or oblong in shape. Numerous black sclerotia were produced
within 5 to 6 days on Potato Dextrose Agar medium. Pathogenicity test was
performed by Koch’s Postulates and fungus isolated was confirmed as
Macrophomina phaseolina.
The isolate of Macrophomina phaseolina produced dark black to brown,
raised colony on potato dextrose agar media. Mycelium was well developed,
hyaline and septate. The fungus produced numerous sclerotia on host and also in
culture medium (PDA). The sclerotia were more or less round with an exception of
oval to irregular in shape. The present finding corroborates with the morphological
characters reported by Singh et al. (1998), such as black, smooth, hard round to
oblong or irregular shape of sclerotia. They measure about 100 microns to 1 mm in
diameter (in culture 50 to 300 microns). However, size is highly variable within an
isolate.
Plate 4.1: Disease symptoms in field
Plate 4.2: Symptoms used for isolation of pathogen
Plates 4.4 Pure culture of Macrophomina phaseolinaon water agar
Plates 4.3 Pure culture of Macrophomina phaseolinaon PDA
Plate 4.5 Sclerotia of Macrophomina phaseolina
4.2 Screening of black gram entries for their reaction to
Macrophomina blight
Forty entries with susceptible check; Barabanki local of black gram
screened for Macrophomina blight under natural field conditions, none of them
were found resistant and moderately resistant. Four entries i. e. KPU 17-1, KPU
17-2, KPU 17-3, KPU 17-4 were found moderately succeptible, nine varieties i. e.
KPU 17-7, KPU 17-9, KPU 17-11, KPU 17-13, KPU 17-25, KPU 17-33, KPU 17-
35, KPU 17-37 and KPU 17-40 were found to be succeptible and twenty seven
entries i. e. KPU 17-5, KPU 17-6, KPU 17-8, KPU 17-10,KPU 17-12, KPU 17-14,
KPU 17-15,KPU 17-16, KPU 17-17,KPU 17-18, KPU 17-19, KPU 17-20,KPU 17-
21,KPU 17-22, KPU 17-23, KPU 17-24, KPU 17-26, KPU 17-27, KPU 17-28,
KPU 17-29, KPU 17-30, KPU 17-31, KPU 17-32, KPU 17-34, KPU 17-36, KPU
17-38 and KPU 17-39 were found to be highly succeptible.
It was cleared that; no entries were found resistant or moderately resistant.
Only 4 entries were moderately succeptible. Rests of the entries were susceptible
and highly susceptible against Macrophomina phaseolina. Deepthi et al. (2014)
revealed that only one entry PKDS-91 was found as moderately resistant to
Macrophomina leaf blight of mungbean. Three entries (OSC-366-I, SSD-2-I and
OSC-79) were recorded as moderately susceptible.
Pandurang (2002) found that screening of thirty-eight black gram entries
was done for testing their resistance to Macrophomina blight. Eleven entries were
found to be moderately resistant to Macrophmina blight. The moderately resistant
entries were K-3, T-9, PU-19, PH-25, LBG-625, PU-30, LAMBG-296, LAMBG-
17, KU-309, KU-314 and KU-303. Rest of the entries were moderately susceptible,
susceptible and highly susceptible against Macrophomina blight.
Zote et al. (1983) also found that out of 19 cultivars screened, none was
completely free of Macrophomina blight however 4 lines were moderately
susceptible.
Table: 4.2 Screening of black gram entries for their reaction to Macrophomina
blight
S. No. Score Reaction Frequency Entries
Distribution
1. 1 Resistant 0 Nil
2. 2 Moderately 0 Nil
Resistant
3. 3 Moderately 4 KPU 17-1, KPU 17-2, KPU 17-3,
susceptible KPU 17-4
4. 4 Susceptible 9 KPU 17-7, KPU 17-9, KPU 17-
11, KPU 17-13, KPU 17-25, KPU
17-33, KPU 17-35, KPU 17-37,
KPU 17-40
5. 5 Highly 27 KPU 17-5, KPU 17-6, KPU 17-
susceptible 8, KPU 17-10, KPU 17-12, KPU
17-14, KPU 17-15, KPU 17-16, KPU 17-17, KPU 17-18, KPU
17-19, KPU 17-20, KPU 17-21, KPU 17-22, KPU 17-23, KPU
17-24, KPU 17-26, KPU 17-27, KPU 17-28, KPU 17-29, KPU
17-30, KPU 17-31, KPU 17-32, KPU 17-34, KPU 17-36, KPU
17-38, KPU 17-39
Total entries – 40
LSI – 4.57
Plate 4.6 Field view of screening trial at different crop stages
4.3 In-vitro evaluation of the Pseudomonas fluorescence isolates for
antagonistic potential against Macrophomina phaseolina.
In vitro antagonistic potential of different isolates of P. fluorescence
wasstudied (Table 4.3 and fig. 4.4) on the mycelial growth of M. phaseolina of
black gram following dual culture method. The mycelial growth was assessed after
7 days of inoculation. Mycelial growth of M. phaseolinaof all the treatments was
significantly less than that of control. Minimum mycelial growth (36 mm) (%
inhibition 60%) of test fungus was observed with isolate P1 followed by P3 growth
(38 mm) and % inhibition (60%), P2 growth (42 mm) and % inhibition (53.6%),
P10 growth (43 mm) and % inhibition (52.3%), P9 growth (44 mm) and %
inhibition (51.2%), P5 growth (45 mm) and % inhibition (50%), P7 growth (45
mm) and % inhibition (50%), P8 growth (48 mm) and % inhibition (46.7%), P4
growth (50 mm) and % inhibition (44.5%) and with highest mycelia growth isolate
P6 growth (65 mm) and % inhibition (27.8%).
Mycelial growth of treatment P1 was at par with P3 and treatment P3 was
at par with P2 and P10.
The work carried out by Pande and Chaube (2003) reported that in vitro
evaluation of Pseudomonas fluorescence isolates resulted in reduction of mycelial
growth of M. phaseolina and the inhibition zone ranged from 1.3 to 22.5 mm in
different isolates on King’s B medium. Similarly work carried out by
Krishnamurthy and Gnanamanickam (1998) reported that Pseudomonas
fluorescence strains recorded antagonistic to P. grisea, R. solani and S. oryzae
were isolated from rice rhizosphere.
Maurya et al. (2014) also found that Pseudomonas fluorescence strain P.f
07 was found most effective with the highest antagonistic activity against three
fungal pathogen and show maximum inhibition of mycelial growth of
Rhizoctoniasolani (68.23%), Fusarium moniliforme (65.45%) and Alternaria
alternata (48.13%).
Table 4.3: Efficacy of Pseudomonas fluorescence isolates againstM. phaseolina
*Average of three replications
Isolates Ave. mycelial growth
(mm)
% growth inhibition
over control
P1 36* 60.0 P2 42 53.6 P3 38 57.8 P4 50 44.5 P5 45 50.0 P6 65 27.8 P7 45 50.0 P8 48 46.7 P9 44 51.2 P10 43 52.3
Control 90.00 -
SE(m)± 1.842
C.D. (5%) 5.438
4.4 In vitroevaluation of fungicides on the mycelial growth of
Macrophomina phaseolina
Six fungicides were tested for their efficacy on Macrophomina phaseolina
in-vitro condition. The chemicals used were Thiram, Propineb,Carbendazim,
Tricyclazole, Carboxin+Thiram, Hexaconazole+Zineb. Incorporated in Potato
Dextrose Agar medium.
The data presented in table 4.4 revealed that all treatments except propineb
were significantly superior over control in checking the mycelial growth of the test
fungus. Tricyclazole, Carboxin+Thiram and Hexaconazole+Zineb completely
inhibited the mycelial growth (0 mm) and % inhibition (100%) followed by
Thiram (60.0 mm) % inhibition (33.4%)), Carbendazim (12.00 mm) % inhibition
(86.66%), and Propineb (90.0 mm)% inhibition (0%).
Mycelial growth of treatment T4 was at par with treatment T5 and T6.
The findings of present study agreed with the findings of other researchers.
Deepthi et al. (2014) revealed that out of different fungicides evaluated under in
vitro conditions against M. phaseolina; vitavax power and penflufen gave
100%inhibition at 500 ppm, while tricyclazole gave 100% inhibition at 1000 ppm
and otherfungicides were less effective. The field evaluation of different fungicides
indicated that vitavax power gave highest seed germination and less pre and post
emergence mortality and yield loss, whereas the sesame seeds treated with vitavax
power alongwith one foliar application of carbendazim was found most effective
for enhancing seed germination and reducing pre and post emergence mortality and
losses in yield of sesame.
Pawar et al. (2014) also revealed that out of eight fungicides tested
Carboxin, Copper oxychloride, Mancozeb, Thiram and Carbendazim were
effective in reducing the growth of M. phaseolina. Sangappa and Mallesh (2016)
revealed that among seven systemic fungicides tested, Benomyl, Hexaconazole,
Thiophanate methyl and Triademefon showed 100 per cent mycelial growth
inhibition.
Table 4.4: Effect of different fungicides on mycelia growth of Macrophomina
phaseolina
Treatment Fungicides
Concentration
in %
Ave. colony
diameter (mm)
Growth reduction
over control (%)
T1. Thiram 1% 60* 33.4
T2. Propineb 2% 90 0
T3. Carbendazim 1% 12 86.66
T4. Tricyclazole 1% 0 100
T5. Carboxin+
Thiram 2% 0 100
T6. Hexaconazole+
Zineb 1% 0 100
T7. Control - 90 -
Se(m)± 0.218
CD at 5% 0.668
*Average of three replications
CHAPTER-V
SUMMARY AND CONCLUSION
The present investigation entitled “STUDIES ON LEAF BLIGHT OF
BLACK GRAM (Vigna mungo (L.) Hepper) OF CHHATTISGARH
PLAINS”were carried out in the laboratories and fields of Departmentof Plant
Pathology, College of Agriculture, Indira Gandhi Krishi Vishwavidyalaya, Raipur
(C.G.). Various aspects statistically analized and results obtained are summarized
below.
Fungal organism was isolated from black gram plants showing typical
symptoms of leaf blight caused by Macrophomina phaseolina. This organism was
purified, identified and pathogenicity test revealed that fungal pathogen was
virulent and produced typical symptoms of leaf blight on black gram.
Out of forty entries screened for resistance against Macrophomina blight,
among all entries except 4 showed susceptible and highly susceptible. Only 4
entries i e. KPU 17- 1, KPU 17- 2, KPU 17- 3, KPU 17- 4 were moderately
succeptible against Macrophomina blight.
In vitro antagonistic potential of different isolates of P. fluorescence
following dual culture method. The mycelial growth was assessed after 7 days of
inoculation. Mycelial growth of M. phaseolina of all the treatments was
significantly inhibit as compare to control. Least mycelial growth (36 mm) term of
inhibition % (60%) of test fungus was observed with isolate P1 followed by P3
growth (38 mm) and % inhibition (60%), P2 growth (42 mm) and % inhibition
(53.6%), P10 growth (43 mm) and % inhibition (52.3%), P9 growth (44 mm) and
% inhibition (51.2%), P5 growth (45 mm) and % inhibition (50%), P7 growth (45
mm) and % inhibition (50%), P8 growth (48 mm) and % inhibition (46.7%), P4
growth (50 mm) and % inhibition (44.5%) and with highest mycelia growth isolate
P6 growth (65 mm) and % inhibition (27.8%).
The data revealed that all treatments except propineb were significantly
superior over control in checking the mycelial growth of the test fungus.
Tricyclazole, Carboxin+Thiram and Hexaconazole+Zineb completely inhibited the
mycelial growth (0 mm) term of inhibition % (100%) followed by Thiram (60.0
mm) % inhibition (33.4%)), Carbendazim (12.00 mm) % inhibition (86.66%), and
Propineb (90.0 mm)% inhibition (0%).
CONCLUSION
The fungus isolated was purified by repeated isolation from the culture plates. The
pathogen was identified on the basis of character of the mycelium and sclerotia.
The mycelium was septate and hyaline, the sclerotia were brown to black in colour,
rounded or oblong in shape.
4 entries i.e. KPU 17- 1, KPU 17- 2, KPU 17- 3, KPU 17- 4 were moderately
succeptible and 9 were shown as susceptible and 27 were shown as highly
susceptible.
All the P. fluorescenceisolates significantly reduced the mycelial growth of
Macrophomina phaseolina over untreated (control) treatment. All the 10 isolates of
P. fluorescenceshowed potential forreduction in mycelial growth. Percent
inhibitions of M. phaseolina ranged from 60 to 27.8 percent.
All the fungicides except propineb were significantly effective in reducing the
radial mycelial growth of Macrophomina phaseolina. Tricyclazole,
Carboxin+Thiram and Hexaconazole+Zineb completely controlled the mycelial
growth and Thiram and Carbendazim was also control the growth of fungus.
SUGGETIONS FOR FUTURE RESEARCH WORK
1) Regular survey works are need to be conducted to assess the severity at regular
intervals. This will help in devising management practices.
2) Need to be development of resistant cultivar against pathogen.
3) Variations within the pathogen also need to be identified for effective resistant
breeding programme.
4) The biological management’s aspects need to be thoroughly investigated.
5) Need based application of chemical fungicides i.e. Tricyclazole, Carboxin+Thiram
and Hexaconazole+Zineb may reduce the diesese when applied at timely interval.
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