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6
SUMMARY AND CONCLUSION
• The study deals with a rapid and reproducible protocol for the
micropropagation of selected explants (nodal and shoot tip) of A. javanica and
A. lanata by direct and indirect organogenesis.
• Optimal regeneration was achieved in MS medium containing 0.5 mg/L SPM
hormone (A. javanica) and 0.5 + 0.5 mg/L BAP + KIN (A. lanata). In vitro
flowering was obtained in the same hormonal concentration.
• Somatic embryos were developed in the 2, 4 D + SPM at 2.0 + 0.5 mg/L. The
embryos were transformed into mature plantlets or sprouts in 1.0 + 0.5 mg/L
BAP + GA3 (A. javanica) and 1.0 + 0.5 mg/L BAP + TDZ (A. lanata)
• Higher concentrations of IBA + IAA (1.0 + 1.0 mg/L) produced maximum
root length and more roots in A. javanica and A. lanata was produced highest
number of roots at 2.0 mg/L IBA.
• The rate of successful acclimatization was 90% for both A. javanica and
A. lanata species.
• About 113 and 126 scorable bands (using four random primers) were observed
in the PCR amplified DNA of A. javanica, and A. lanata.
• About 98 monomorphic and 15 polymorphic bands were reported in A.
javanica with lowest percentage of polymorphic variations (13.27 % in all
samples of in vivo, in vitro leaf and callus).
• Percentage of polymorphic changes in the samples of A. lanata were 10.56 %
(110 bands were monomorphic and 13 were polymorphic bands).
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• Jaccard’s similarity and distance coefficient values have shown similarity of
1.000 and distance of 0 scales among the samples.
• Based on the results of monomorphic bands occurred in the DNA samples, the
uniform distribution of secondary metabolites contents (for in vivo, in vitro
leaf and callus materials) is ensured.
• Little/few polymorphic variations occurred in gDNA within the samples
indicates the improved secondary metabolites contents in plants.
• The antimicrobial activity of A. javanica showed that most of the extracts
exhibited broad spectrum (higher to moderate range) of inhibition against the
tested bacterial and fungal pathogens.
• Highest inhibition was observed in the callus and in vitro leaf methanolic
extracts against S. aureus, E. faecalis, B. subtilis, and S. typhi followed by
chloroform, acetone and ethyl acetate extracts. A. niger and Fusarium was
found to be inhibited by the extracts tested.
• Callus and in vitro leaf extracts (ethyl acetate, methanol, and chloroform
solvents) of A. lanata was able to inhibit the tested pathogens i.e. S. aureus,
S. typhi, B. subtilis, E. coli, S. boydii, and P. vulgaris. Similarly, all the
extracts were found to have some inhibitory effects against the fungus namely
C. albicans and A. niger.
• The MIC values of methanolic extract of plant samples were found to be in the
range of 18 – 22 µg/mL (in vivo extracts), 15 – 22 µg/mL (in vitro extracts)
and 16 – 24 µg/mL (callus extracts) against the tested pathogens.
• MIC of methanolic extracts of A. lanata was reported in the range of 19 – 28
µg/mL (in vivo), 21 – 25 µg/mL (in vitro) and 12 – 17 µg/mL (callus). Colony
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count of MBC plate from the respective MIC tubes also show clear or
negligible amount of colonies.
• Fractions separated from A. javanica and A. lanata by TLC was tested against
both Gram positive and negative organisms, (S. aureus and S. typhi). Fractions
of AJ-IL-F1, AJ-FL-F1, AJ-CA-F1 were found to inhibit the growth of
organisms.
• The qualitative phytochemical analysis of results shows the presence of
flavonoids, phenolic contents, tannins and carbohydrates in all the extracts of
tested plants. Alkaloids, saponins, terpenoids and fats and oils were found to
be present in most of the extracts of both plants.
• The methanol and aqueous extracts of A. javanica were reported to contain
higher amount of phenolics (196.68 ± 10.45 mg GAE/g), tannins (116.08 ±
3.71 mg GAE/g), flavonoids (177.54 ± 1.8 mg RE/g) and carbohydrates (139.0
± 5.1 mg GE/g).
• The extracts of A. lanata extracts was reported to contain higher amount of
phenols (193.98 ± 10.58 mg GAE/g), tannins (109.41 ± 9.42 mg GAE/g),
flavonoids (151.45 ± 6.97 mg RE/g) and carbohydrates (58.60 ± 3.6 mg GE/g)
from in vitro leaf extracts followed by in vivo and callus extracts.
• Antioxidant activity was tested by different assays. The DPPH assay results
showed lower IC50 values (87.02, 113.45, 71.54 µg/mL) which were recorded
in the methanol extracts (in vivo, in vitro leaf and callus) followed by acetone
and aqueous extracts in A. javanica.
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• The antioxidant potential of A. lanata recorded least IC50 values (53.77 and
64.42 µg/mL) from in vivo and in vitro leaf methanol extracts followed by
aqueous extracts (DPPH assay).
• The total antioxidant potential of A. javanica was tested by
phosphomolybdenum, ferric reducing power and metal chelation assay. It was
observed that the methanolic extracts possessed higher antioxidant potential
followed by aqueous and ethyl acetate extracts.
• The in vitro leaf has higher antioxidant content based on FRAP and
phosphomolybdenum assay. In vivo leaf extracts has higher antioxidant
potential in metal chelation assay. Similarly, callus extracts also possessed
equal amount of total antioxidant content when compared with in vivo and in
vitro plants.
• Total antioxidant activity of A. lanata was tested by FRAP,
phosphomolybdenum and metal chelation assay which showed higher activity
in methanol extracts (in vitro and in vivo leaf extracts and callus extracts). Rest
of the solvent extracts (in vivo, in vitro leaf and callus) showed considerable
activity. Among them, the in vitro leaf and callus extracts were found to have
promising antioxidant property.
• The methanolic extract having higher antioxidant potential (by in vivo, in vitro
leaf and callus extracts) was further tested for anticancer activity on MCF – 7
cell line (cytotoxic potential) which showed better anticancer activity than the
commercial compound (5 - FU) with least IC50 values (AJ-FL = 114.43,
AJ-IL = 11.89, AJ-CA = 27.18 µg/mL) from A. javanica. The IC50 values of
A. lanata was recorded as AL-FL = 43.89, AL-IL = 167.77, and AL-CA =
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22.45 µg/mL. Further DNA fragmentation assay results showed the digestion
of DNA damaged during the process of apoptosis.
• Preparative TLC of in vivo, in vitro leaf and callus methanol extracts produced
6 fractions (in vivo – 03, in vitro – 02, and callus 01) in A. javanica and
9 fractions (4 - in vivo leaf, 3 - in vitro leaf and 2 - callus) from A. lanata
extracts.
• Based on the Rf value, fractions separated from the extracts is found to belong
to the polyphenolic group of compounds.
• The fractions also confirmed the presence of phenolic and flavonoids
compounds as interpreted by the peaks obtained in the HPLC chromatogram
along with the standards (gallic acid, epicatechin, catechin, quercetin, vanillin,
chlorogenic acid, ferulic acid and resveratrol). Contents of the fractions were
quantified and expressed as the amount of standard equivalents.
• During UV spectral analysis, the absorbance peak was noticed at 248 nm (for
in vivo fraction) and 264 nm (for in vitro and callus fractions) for A. javanica.
• A. lanata also showed the highest absorbance peak at 283 nm (in vivo
fraction), 240 nm (in vitro fractions) and 268 nm (callus fraction) by UV
spectral analysis.
• GC – MS analysis of A. javanica showed 19 compounds from in vivo leaf and
9 from in vitro and callus extracts. About 16 compounds were identified (GC –
MS) in A. lanata, in vivo leaf and 6 compounds from in vitro and callus
extracts.
• The chemical library analysis results showed that the stigmasterol compound
(a polyphenolic group) was found in all the extracts and Methyl-methyl
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deconate was the abundant compound present in the tested extracts, followed
by Hentriacontane, a major compound present in both plant extracts.
• Functional groups of the compounds were found and the presence of
C-H, C=O, O-N, -NH3, stretching, deformation and bending’s along with
aldehyde, ketone, amide and peptide linkages were found in the
identified compounds of A. javanica and A. lanata. Similar type of functional
group and chemical bonds were also reported in isolated fractions from both
plants.
• Based on the mass spectral analysis (by LC – MS) for the isolated
fractions, the molecular weight as follows (AJ FL F1 – 313.23, AJ IL F1 –
251.25, AJ CA F1 – 299.23, AL FL F1 – 343.22, AL IL F1 – 251.25, and
AL CA F1 – 439.18 kDa) from A. javanica and A. lanata respectively is
recorded.
• In a nutshell it is stated that micropropagation of A. javanica, anticancer
activity of in vivo, in vitro leaf and callus materials of both plants (A. javanica
and A. lanata) against human breast cancer cell line and LC – MS
spectral analysis of the fractions of both plants are reported for the first time.
• Based on the outcome of this study, it is confirmed that, the in vitro
leaf and callus materials from both the test plants can be efficiently used
for mass production of pharmaceutical drugs compared to the in vivo test
plant.
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SUGGESTIONS AND FUTURE RECOMMENDATIONS
• Purification and in vivo trails (with animal model) for the isolated fractions.
• Analysis of Nuclear Magnetic Resonance of the compounds for
identification of its molecular structure i.e., elemental analysis.
• Molecular docking of the isolated compounds for prediction with
commercial compounds.
• Pharmacological aspects of isolated compounds.
with (Soliman, infections, swelling (Garg et al., 1975) 5-methylmellein
matted 2006) 1980; Qasim et al., 2013). (Claydon et al., 1985),
hairs Leaves Kaempferol-3-O-β-D-
Aerva javanica (Burm.f.) Juss. (Khare, Ulcer in animals, cattle fodder glucopyranosyl-(1→2)-α-
ex Schult. 2007) (Qureshi and Bhatti, 2009) L-rhamnopyranoside-7-O-
Seeds α-L-rhamnopyranoside
1.1. Description of plants
1.9.1. Description of Aerva javanica
Plant name/ Taxonomic
Botanical description Ethnobotanical/ pharmacological
position Habit Leaves Flowers aspects Phytochemical nature
Kingdom : Plantae
Division : Magnoliophyta
Class : Magnoliopsida
Order : Caryophyllales
Family : Amaranthaceae
Genus : Aerva
Species : javanica (Burm. f.)
Juss. ex Schult
Perennial
herb,
Grows
up to
1.6m
(Khare,
2007)
Pale green,
20 – 40mm
long,
Alternate,
Lanceolate,
Shortly
petiolate,
Covered
White woody,
Raceme
inflorescence,
Flowering
period
January to
May
Roots and flowers
Rheumatism, Kidney problems,
tooth ache, wound bleeding, control
eye disease, (Mossa et al., 1987;
Abbas et al., 1992; Ghazanfar,
1994)
Whole plant
Dysentery, gonorrhoea, cutaneous Head ache, rheumatism (Srivivan
and Reddy, 2008)
Pharmacological properties
Antiplasmodial,
(Joanofarc and Vamsadhara, 2003;
Cytogentical and cytotoxic
(Soliman, 2006; Fatimi, 2007),
antihyperglycaemic (Reddy and
Reddy, 2009), antidiarrhoeal
properties (Ahmed et al., 2010)
Antimicrobial (Sharif et al., 2011).
Flavonoids,
steroids, triterpenoids and
carbohydrates (Reddy
and Reddy 2009; Sharif et
al., 2011)
Isolated compounds
Apigenin 7-O-glucuronide
(Kenneth and Lawrence (Kucukislamoglu et al.,
2000), Isoquercetrin
(Guvenalp, and Demirezer,
2005), 7-(1’hydroxyethyl)-
2-(2″-hydroxyethyl)-3,4-
dihydrobenzopyran
(Donnell et al., 2006),
2-hydroxy-3-O-β-
primeveroside naphthalene-
1,4-dione (Ozgen et al.,
2009),
1.9.2. Description of Aerva lanata Botanical description Ethno botanical/ pharmacological
Plant name/ Taxonomic position
Habit Leaves Flowers aspects Phytochemical nature
Aerva lanata (L.) A. L. Juss. ex
Schultes
Kingdom : Plantae
Division : Magnoliophyta
Class : Magnoliopsida
Order : Caryophyllales
Family : Amaranthaceae
Genus : Aerva
Species : lanata (L.) A. L. Juss. ex
Schultes
Prostrate,
decumbent,
sometimes
erect herb,
30-60 cm in
height,
woolly
tomentose
(Khare 2007;
Rajesh et al.,
2011)
Leaves are
simple,
alternate
and short,
petiolated,
densely
tomentose
Small and
hairy
white
colour,
sessile,
bisexual,
clustered
spikes
(Rajesh et
al., 2011)
Whole plant
Cough, sore throat,
wounds, cataract of
bladder, animal
constipation nasal
bleeding, fractures,
spermatohorrhoea,
cough, scorpion stings
(Mukerjee et al., 1984;
Sikarwar and Kaushik,
1993; Girach et al.,
1994), burning sensation
during urination
(Venkataramana, 2008).
Flowers and Roots
Headache, scabies,
jaundice, diarrhoea and
kidney disorders (Bedi
and Patel, 1978; Muthu
et al., 2006;
Soundararajan et al.,
2006; Khare, 2007).
Alkaloids, flavonoid, steroid,
terpenoids, phenolic compounds,
tannins and carbohydrates (Aiyar
et al., 1973; Chandra and Sastry
1990)
Isolated compounds
Alkaloids
Canthin-6-one beta-carboline,
aervine (10-hydroxycanthin- 6-
one), methylaervine (10-
methoxycanthin-6-one),
aervoside (10-β-
dglucopyranosyloxycanthin-6-
one) and aervolanine (3-(6-
methyoxy-β-carbolin-1-yl)
propionic acid) (Zapesochnaya,
1992),
Contd...
Leaves
Eye complaints, diarrhoea,
treatment against guinea worm
Roots
Snake bite, hepatitis, urinary
strangury (Yamunadevi et al.,
2011).
Pharmacological properties
Diuretic (Chopra et al., 1956;
lithiatic, urolithiasis (Rao,
1985), Udupihille and Jiffry,
1986), antitumor activities
(Zapesochnaya et al., 1992).
Diabetes, anthelmintic,
demulcent lithiasis (Kirtikar
and Basu, 1996), anti-
inflammatory (Vetrichelavan
et al., 2000), antimicrobial and
cytotoxic (Dulay, 2002)
hepatoprotective and nephro
protective (Shirwaikar et al.,
2004).
Glycosides
Kaempferol 3 –
rahamnogalactoside,
kaempferol 3 – 6” p –
coumaryl O – glucoside
(Mallabev et al., 1989)
Flavonoids
Tiliroside, coumaryl
tiliroside, isorhamnetin
(Mallabev et al., 1989).
Table 16: Shooting and in vitro flowering response of A. javanica in different concentrations of cytokinins and polyamines in direct
and indirect organogenesis
Node explants Regenerat Shoot tip explants Regenerat In vitro
Hormone
Concentration
(mg/L)
Shoot length
(cm)
No. of Multiple
shoot-1 explant
ion rate
(%) Shoot length
(cm)
No. of Multiple
shoot-1 explant
ion rate
(%)
Flowerin
g (%)
Basal 0 - - - - - - -
BAP 0.5 2.15 ± 0.36ab 3.0 ± 1.06 ab 57.14 2.27 ± 0.51 ab 2.14 ± 0.91 ab 92.8 -
1.0*
4.80 ± 0.92abc
5.71 ± 1.03abc
100 3.67 ± 0.61 ab
4.57 ± 2.09abc
92.85 -
1.5 2.83 ± 0.89a
2.78 ± 1.08a
85.71 2.83 ± 0.24a
2.50 ± 0.50a
92.85 -
2.0 1.30 ± 0.14cd
2.14 ± 0.14cd
85.71 1.51 ± 0.18cd
2.35 ± 0.61cd
82.71 -
BAP + KIN 0.5 + 0.5 1.95 ± 0.42cd 1.42 ± 0.49cd 35.17 1.51 ± 0.47cd 0.42 ± 0.49e 42.85 -
1.0 + 0.5 2.38 ± 0.21a 1.92 ± 0.96cd 35.17 2.47 ± 0.41ac 1.0 ± 0e 57.17 -
1.5 + 0.5 1.22 ± 0.15cd
2.28 ± 1.09ac
85.71 1.08 ± 0.46cd
0.14 ± 0.51e
85.75 -
2.0 + 0.5 1.24 ± 0.14cd
1.92 ± 1.16cd
85.71 1.53 ± 0.28cd
3.35 ± 1.44 85.71 -
2.5 + 1.0 2.50 ± 0.37a
2.35 ± 1.39ac
78.57 2.60 ± 0.25 ac
3.0 ± 0 ab
71.42 -
3.0 + 1.0 1.50 ± 0.70cd 1.78 ± 0.67cd 85.71 1.65 ± 0.23cd 2.42 ± 0.62ab 92.85 -
4.0 + 0.5 2.25 ± 0.13a 1.57 ± 0.49cd 85.71 2.49 ± 0.15ac 2.49 ± 0.15ab 92.85 -
BAP + SPM 1.0 + 0.5 3.81 ± 1.04ab
2.0 ± 0.65a
92.85 3.57 ± 0.15ab
3.0 ± 0ab
78.57 -
SPM 0.3#
2.76 ± 1.59a
2.42 ± 1.67ab
92.85 4.16 ± 0.54abc
3.0 ± 0ab
100 -
0.4 3.59 ± 1.22ab
3.42 ± 0.90abc
85.71 2.75 ± 0.16a
3.0 ± 0ac
85.71 -
0.5# 3.70 ± 1.77ab 3.0 ± 1.77ac 85.71 4.23 ± 0.27abc 3.57 ± 1.04ab 100 57.14
BAP + GA3^ 1.0 + 0.2 3.41 ± 1.04ab 3.81 ± 1.03ab 92.85 3.14 ± 0.55ac 2.82 ± 1.40ac 64.2 -
1.0 + 0.5 3.38 ± 0.37ab
3.2 ± 0.71ab
85.71 3.0 ± 0.19ac
2.70 ± 0.60c
64.2 -
BAP + TDZ^
1.0 + 0.2 3.40 ± 0.71ab
3.0 ± 0.10ac
78.57 3.0 ± 0.40ab
3.84 ± 1.28ab
85.71 -
1.0 + 0.5 3.01 ± 0.54ab 3.1 ± 1.06ac 78.57 3.0 ± 0.40ab 3.74 ± 1.81ab 85.71 -
Values in the table are the mean ± SE of triplicates, n=14 explants, Superscript in the each row carrying different letters are
significantly different at P<0.05 or 0.01, *-Significance at 0.05, # - Significance at 0.01 by Newman-Keuls Multiple Comparison Test.
^ - hormones used in shoot elongation of indirect organogenesis.
Figure 3: In vitro regeneration and embryogenesis of A. javanica in different concentration of cytokinins and spermidine, A. Regenerated
shoot tip explant, B. nodal explant, C. callus regeneration, D. multiple shooting, E. in vitro flowering, F. shoot elongation, G. in vitro roots,
H. matured plants (hardened).
Rate of callus Callus morphology
formation
Rate of
embryo
Frequency of
embryos
No. of shoots/embryo
cultures
++ Pale brown fragile - - -
++ Pale brown fragile - - -
++ Pale yellow fragile - - -
+++ Pale yellow fragile - - -
++ Yellow with pinkish
soft
+
10.33 ± 0.47ab
-
++ Pale yellow green soft + 11.33 ± 0.47ab
-
++++ Yellow green with -
pinkish soft +++ 88.40 ± 2.62cd
Table 17: Embryogenesis and indirect organogenesis from embryos of A. javanica callus
Hormone Concentrations
2, 4 - D 0.5
1.0
2.0
3.0
2, 4 - D +
NAA 2.0 + 0.5
2.0 + 1.0
2, 4 - D + SPM 2.0 + 0.5*
2, 4 – D +
ABA 0.5 + 0.2
* +++
Pale yellow green hard
++ 55.47 ± 1.04d
-
cd ab
0.5 + 0.5* +++ Pale yellow green with
pinkish hard
+++ 88.46 ± 2.62 1.2 ± 0.47
2, 4 - D + BAP # ++++ +++ 88.40 ± 2.62
cd 13.8 ± 1.20
a
+ SPM 2.0 + 2.0 + 0.5
Green
+ rate of callus and embryo development, - no embryo or shoot development. Values in the table are the mean ± SE of triplicates, n=14
explants, Superscript in the each row carrying different letters are significantly different at P<0.05 or 0.01, *-Significance at 0.05,
# - Significance at 0.01 by Newman-Keuls Multiple Comparison Test
Figure 4a: Embryogenesis of A. javanica at different concentrations of auxin and
spermidine, A. Greenish brown callus, B. embryo forming callus, C. matured
embryos, D. regeneration from embryo cells, E. sprouts/plantlets F. development of
multiple sprouts.
Figure 4b: Nature of different forms of callus in A. javanica, A. pale yellow green
callus, B. brown callus, C. yellow callus, D. greenish callus (Non embryo forming
cells)
Figure 6: In vitro regeneration and embryogenesis of A. lanata, A. regeneration of nodal explants, B. shoot tip explants, C. multiple
shooting, D. shoot elongation, E. in vitro flowering, F. in vitro rooting, G. acclimatized plants.
Table 19: Shooting and in vitro flowering response of A. lanata different concentrations of cytokinins and polyamines in direct and
indirect organogenesis
Hormones
Concentration
of hormones
L Shoot tip explants
Regeneration
Node explants Regener
ation
In vitro
Flowering
(mg/L) Shoot length
(cm)
No. of Multiple shoot
-1 explant
rate (%) Shoot length (cm)
No. of Multiple shoot
-1 explant
rate (%) (%)
Basal MS basal - - - - -
BAP 0.5 2.72 ± 0.41ab
3.10 ± 0.32bc
64.2 1.21 ± 0.31bc
1.33 ± 0.31bc
46.8 -
0.75 4.50 ± 0abc
3.94 ± 0.18b
64.2 1.25 ± 0.40bc
3.9 ± 0.10b
57.1 -
1.0* 5.33 ± 0.41abc
5.76 ± 0.91abc 85.7 4.83 ± 1.4abc
3.98 ± 1.19b 92.8 78.5
1.5 3.20 ± 0.55b 3.42 ± 0.6b
78.5 3.12 ± 1.4bc 1.24 ± 0.6bc
85.7 -
2.0 4.0 ± 0.63abc 4.0 ± 0.6ab
71.4 3.40 ± 1.2bc 2.81 ± 1.41bc
78.5 36.7
BAP + KIN 0.5 + 0.5# 5.76 ± 0.85
abc 4.91 ± 1.42
abc 92.8 4.93 ±0.84
abc 5.81 ± 0.1
abc 100 92.8
0.5 +0.75 2.72 ± 0.34ab
2.62 ± 0.11b
64.2 1.77 ± 0.33bc
3.26 ± 0.61bc
78.5 -
0.5 + 1.0 3.61 ± 0.24b 3.52 ± 0.6b
71.4 2.81 ± 0.22b 3.05 ± 0.14b
78.5 -
1.0 + 1.0 3.54 ± 0.61b 3.52 ± 0.6b
78.5 3.35 ± 2.51b 3.0 ± 0b
71.4 -
BAP + SPM 1.0 + 1.0 2.21 ± 1.04ab 2.0 ± 1.51ab
92.8 2.29 ± 1.5c 2.40 ± 0.47b
85.7 -
SPM 0.3 2.0 ± 0ab
2.42 ± 0.61ab
78.4 2.16 ± 3.58c 1.10 ± 0.21
bc 54.7 -
0.4 1.59 ± 1.53bc
1.28 ± 1.96bc
54.7 1.50 ± 2.16bc
1.8 ± 0bc
54.7 -
0.5 2.70 ± 1.27ab 2.90 ± 1.21ab
64.2 2.32 ± 2.71c 2.25 ± 0.44b
64.2 -
BAP + GA3^ 1.0 + 0.2 2.11 ± 2.0ab 2.81 ± 1.0ab
71.4 2.14 ± 1.55abc 2.32 ± 0.40bc
64.2 78.4
1.0 + 0.5* 2.33 ± 0.3
ab 2.42 ± 0.7
ab 64.2 1.96 ± 1.19
bc 1.75 ± 1.60
bc 64.2 -
BAP + TDZ^ 1.0 + 0.2 2.10 ± 1.71ab
2.20 ± 0.61ab
57.1 2.92 ± 0.65abc 2.84 ± 1.28
ac 64.2 -
1.0 + 0.5*
2.51 ± 1.54ab
2.81 ± 1.46ab
85.7 2.87 ± 1.40abc
2.74 ± 1.81ac
85.7 71.4
Values in the table are the mean ± SE of triplicates, n=14 explants, Superscript in the each row carrying different letters are
significantly different at P<0.05 or 0.01, *-Significance at 0.05, # - Significance at 0.01 by Newman-Keuls Multiple Comparison Test
^ - hormones used in shoot elongation of indirect organogenesis
Figure 7: Embryogenesis of A. lanata in different concentration of auxins and spermidine A. Embryo forming callus, B. greenish embryo
cells, C. yellow brown embryo cells, D. yellow embryo cells, E. initiation of sprouts from yellow brown embryo cells, F. sprout maturation.
abc
Table 20: Embryogenesis and indirect organogenesis from A. lanata No. of
Hormone Concentrations Rate of callus
formation Callus morphology
Rate of
embryo
Frequency of embryos
shoots/embryo
cultures
2, 4 – D 0.5 +++ Green soft - - -
1.0 +++ Greenish yellow soft - - -
1.5 +++ Yellow hard - - -
2.0 +++ Pale yellow soft - - -
2, 4 – D + NAA 0.5 + 0.5 ++ Pale yellow soft - - -
0.5 + 1.0 ++ Pale yellow soft - - -
0.5 + 1.5 +++ Pale yellow green hard - - -
0.5 + 2.0 ++++ Pale yellow green hard - - -
2, 4 - D + SPM 2.0 + 0.5*
+++
Yellow green with pinkish -
soft +++ 88.40 ± 2.62abc
ab
2,4 – D + ABA 0.5 + 0.2 +++ Pale yellow green hard ++ 45.74 ± 1.94 -
0.5 + 0.5#
++++ Pale yellow green with
pinkish hard
+++ 87.56 ± 1.28 -
2,4 – D + BAP + # +++ 92.40 ± 1.62
abc 58.82 ± 2.20
ab
SPM 2.0 + 2.0 + 0.5
++++ Green
+ rate of callus and embryo development, - no embryo or shoot development. Values in the table are the mean ± SE of triplicates, n=14
explants, Superscript in the each row carrying different letters are significantly different at P<0.05 or 0.01, * - Significance at 0.05,
# - Significance at 0.01 by Newman-Keuls Multiple Comparison Test
Table 21: In vitro rooting of A. lanata in different concentrations of auxins
Hormones
Concentration
of hormones
(mg/L)
Mean of root
length (cm)
Number of
multiple root/
stem
Rate of roots
formation (%)
IBA 0.5 2.7 ± 0.6c
2.5 ± 0.7c
50
1.0*
3.4 ± 0.9abc
3.3 ± 0.9ab
64.2
1.5 4.8 ± 1.1bc
5.0 ± 1.2abc
64.2
2.0#
6.8 ± 1.4abc
7.6 ± 0.4abc
100
IBA + IAA 0.5 + 0.5 4.1 ± 0.7bc
3.8 ± 0.6bc
64.2
0.5 + 1.0*
3.3 ± 0.5abc
2.9 ± 0.7bc
42.8
0.5 + 1.5*
3.2 ± 0.3abc
2.7 ± 0.5c
42.8
0.5 + 2.0*
3.2 ± 0.2abc
3.2 ± 0.5abc
42.3
IBA + NAA 0.5 + 0.5 2.6 ± 0.9c
2.5 ± 0.6c
57.14
0.5 + 1.0 3.0 ± 0.7bc
2.8 ± 0.7c
57.14
0.5 + 1.5 3.0 ± 0.8bc
3.0 ± 0.7bc
42.8
0.5 + 2.0 3.3 ± 0.4ab
3.2 ± 0.4ab
42.8
Values in the table are the mean ± SE of triplicates, n=14 explants, Superscript in
the each row carrying different letters are significantly different at P<0.05 or 0.01,
*-Significance at 0.05, # - Significance at 0.01 by Newman-Keuls Multiple
Comparison Test
Figure 8: In vitro regenerated plants in polycups (after 4 weeks of acclimatization)
Plants
Primer
Total no. of
scorable
bands
No. of
monomorphic
bands
No. of
polymorphic
bands
Oligo 01 27 23 4
Oligo 02 28 25 3
A. Oligo 03 30 26 4
javanica Oligo 04 28 24 4
Table 24: RAPD-PCR amplification products of DNA extracted from different
samples (in vivo, in vitro leaf and callus) of A. javanica
% of
polymorphism
Overall 98
total 113
14.81
10.71
13.33
14.28 15 13.27
% of polymorphism = no. of polymorphic bands/ total no. of scorable bands X 100.
Table 25: Distribution and size of polymorphic bands of different samples (in vivo,
in vitro leaf and callus) materials of A. javanica
Primer Size of polymorphic
band (kb)
Distribution of polymorphic bands
In vivo leaf In vitro leaf callus
Oligo 01
Oligo 02
Oligo 03
Oligo 04
5.0 - + +
2.5 + - +
2.0 - - -
1.5 - - -
1.0 - - -
500 bp - - -
5.0 + - -
2.5 - - +
2.0 - - +
1.5 - - -
1.0 - - -
500 bp - - -
5.0 - - +
2.5 - - -
2.0 - - -
1.5 - - -
1.0 + - +
500 bp - + -
5.0 - - -
2.5 - - +
2.0 - - -
1.5 - - -
1.0 - + +
500 bp + - -
+ Present, – Absent of polymorphic band
Figure 10: RAPD Profile of A. javanica DNA using different primers (Oligo 01, Oligo 02, Oligo 03, and Oligo 04), Sample I – in vivo
leaf, sample II – in vitro leaf, sample III – callus, L – Marker
Table 26: Similarity Matrix computed with Jaccard’s coefficient of A. javanica
Sample 1 Sample 2 Sample 3 Sample 1R Sample 2R Sample 3R
Sample 1 1 0.204 0.416 1.000 0.204 0.416 Sample2 1 0.400 0.204 1.000 0.400
Sample 3 1 0.416 0.400 1.000
Sample 1R 1 0.204 0.416
Sample 2R 1 0.400
Sample 3R 1
Table 27: Distance Matrix based on Jaccard’s coefficient of A. javanica
Sample 1 Sample 2 Sample 3 Sample 1R Sample 2R Sample3R
Sample 1 0 0.216 0.190 0.000 0.216 0.190
Sample 2 0 0.200 0.216 0.000 0.200
Sample 3 0 0.190 0.200 0.000
Sample 1R 0 0.216 0.190
Sample 2R 0 0.200
Sample 3R 0
Table 28: Distance matrix based on RMSD coefficient of A. javanica
Sample 1 Sample 2 Sample 3 Sample 1R Sample 2R Sample3R
Sample 1 0 0.313 0.207 0.000 0.313 0.207
Sample 2 0 0.192 0.313 0.000 0.192
Sample 3 0 0.207 0.192 0.000
Sample 1R 0 0.313 0.207
Sample 2R 0 0.192
Sample 3R 0
Sample 1R - 3R – Repeat of Sample 1 – 3, Sample 1 = in vivo leaf;
Sample 2 = in vitro leaf; Sample 3 = in vitro callus
Figure 11: Phylogenetic Tree Analysis (Phenogram) of A. Javanica, Sample 1 = in
vivo plant leaf, Sample 2 = in vitro plant leaf, Sample 3 = in vitro callus.
band (bp) In vivo plants In vitro plants callus
600 - + -
RBA 10 300 - - +
200 - + -
600 + - -
RBA 13 300 - + +
200 - - +
OPS 01 600 - + -
300 + - +
200 - - -
OPS 12 600 - - -
300 + - -
200 - + +
Table 29: RAPD-PCR amplification products of DNA extracted from different
samples (in vivo, in vitro leaf and callus) of A. lanata using four random primers
Plants Primer
Total no. of
scorable
bands
No. of
monomorphic
bands
No. of
polymorphic
bands
% of
polymorphism
A. lanata
RBA 10 28 25 3 10.71
RBA 13 34 30 4 11.76
OPS – 01 33 30 3 9.09
OPS – 12 28 25 3 10.71
Overall total 123 110 13 10.56
% of polymorphism = no. of polymorphic bands/ total no. of scorable bands X 100.
Table 30: Distribution and size of polymorphic bands of different samples from A. lanata
Primer Size of polymorphic Distribution of polymorphic bands
+ Present – Absent of polymorphic band
Figure 12: RAPD Profile of A. lanata DNA using different primers, S1 – in vivo leaf, S2 – in vitro leaf, S3 – callus, M – Marker
Table 31: Similarity matrix computed with Jaccard’s coefficient of A. lanata
Sample 1 Sample 2 Sample 3 Sample 1R Sample 2R Sample3R
Sample 1 1 0.414 0.237 1.000 0.414 0.237
Sample 2 1 0.200 0.414 1.000 0.200
Sample 3 1 0.237 0.200 1.000
Sample 1R 1 0.414 0.237
Sample 2R 1 0.200
Sample 3R 1
Table 32: Distance matrix based on Jaccard’s coefficient of A. lanata
Sample 1 Sample 2 Sample 3 Sample 1R Sample 2R Sample3R
Sample 1 0 0.586 0.763 0.000 0.586 0.763
Sample 2 0 0.800 0.586 0.000 0.800
Sample 3 0 0.763 0.800 0.000
Sample 1R 0 0.586 0.763
Sample 2R 0 0.800
Sample 3R 0
Table 33: Distance matrix based on RMSD coefficient of A. lanata
Sample 1 Sample 2 Sample 3 Sample 1R Sample 2R Sample 3R
Sample 1 0 0.636 0.831 0.000 0.636 0.831
Sample 2 0 0.816 0.636 0.000 0.816
Sample 3 0 0.831 0.816 0.000
Sample 1R 0 0.636 0.831
Sample 2R 0 0.816
Sample 3R 0
Sample 1R - 3R – Repeat of Sample 1 – 3, Sample 1 = in vivo plant leaf; Sample 2 = in vitro
plant leaf; Sample 3 = in vitro callus
Figure 13: Phylogenetic Tree Analysis (Phenogram) of A. lanata, Sample 1 = in vivo
leaf, Sample 2 = in vitro leaf, Sample 3 = in vitro callus.
Table 34: Antibacterial activity of different solvents extracts A. javanica (in vitro, field leaf and callus)
Org name Method Hexane Chloroform Ethyl acetate
IL FL CA C IL FL CA C IL FL CA C S. aureus WELL 11.0±0 12.0±0 10.0±0 21.0±1.0 9.0±0 8.0±0 9.0±0 21.0±1.0 12.0±0 12.0±0 12.0±0 26.0±1.0
DISC 9.0±0 8.0±0 8.0±0 10.0±1.0 9.0±1.0 9.0±1.0 9.0±1.0 10.0±1.0 7.0±0 6.0±0 12.0±0 14.0±1.0
S. typhi WELL - 11.0±1.0 12.0±1.0 21.0±1.0 - - - 11.0±1.0 8.0±0 9.0±0 9.0±0 26.0±1.0
DISC - - - 21.0±1.0 9.0±1.0 9.0±1.0 9.0±0 13.0±1.0 9.0±1.0 9.0±0 9.0±0 14.0±1.0
C. diphtheriae WELL 10.0±1.0 8.0±1.0 8.0±1.0 26.0±1.0 12.0±1.0 12.0±0 14.0±1.0 26.0±1.0 11.0±0 11.0±1.0 11.0±1.0 26.0±1.0
DISC 9.0±0 6.0±0 8.0±1.0 21.0±1.0 6.0±0 8.0±0 8.0±0 17.0±1.0 8.0±0 9.0±0 12.0±1.0 14.0±1.0
P. vulgaris WELL 11.0±0 9.0±1.0 8.0±0 26.0±1.0 8.0±1.0 12.0±0 12.0±0 16.0±1.0 12.0±0 12.0±0 11.0±1.0 26.0±1.0
DISC 7.0±0 7.0±0 9.0±1.0 12.0±1.0 9.0±1.0 10.0±0 9.0±1.0 15.0±1.0 8.0±0 8.0±0 8.0±0 14.0±1.0
V. vulnificus WELL 9.0±1.0 9.0±1.0 - 26.0±1.0 8.0±1.0 8.0±0 9.0±1.0 26.0±1.0 - - - 26.0±1.0
DISC 7.0±1.0 7.0±0 8.0±0 12.0±1.0 8.0±0 8.0±0 8.0±0 15.0±1.0 - - - 10.0±1.0
S. dysenteriae WELL 8.0±0 12.±0 7.0±0 15.0±1.0 9.0±1.0 13.0±1.0 8.0±0 15.0±1.0 12.0±0 9.0±1.0 9.0±0 26.0±1.0
DISC - - - 17.0±1.0 8.0±0 8.0±0 7.0±0 15.0±1.0 10.0±0 9.0±1.0 10.0±0 12.0±1.0
E. coli WELL 8.0±0 13.0±1.0 8.0±0 21.0±1.0 9.0±1.0 13.0±0 9.0±1.0 21.0±1.0 - - - 21.0±1.0
DISC 6.0±0 8.0±0 12.0±1.0 16.0±1.0 7.0±0 7.0±0 7.0±0 15.0±1.0 - - - 16.0±1.0
E. faecalis WELL 11.0±1.0 12.0±0 - 26.0±1.0 10.0±1.0 12.0±0 12.0±0 26.0±1.0 14.0±0 13.0±1.0 12.0±0 26.0±1.0
DISC 6.0±0 6.0±0 6.0±0 12.0±1.0 - - - 15.0±1.0 9.0±1.0 8.0±1.0 12.0±0 12.0±1.0
L. acidophilus WELL 8.0±0 11.0±0 12.0±0 16.0±1.0 11±1.0 12.0±0 12.0±0 26.0±1.0 8.0±0 10.0±1.0 12.0±0 16.0±1.0
DISC 7.0±0 8.0±0 8.0±0 12.0±1.0 6.0±0 8.0±0 12.0±0 12.0±1.0 9.0±0 7.0±0 7.0±0 25.0±1.0
P. aeruginosa WELL 12.0±1.0 12.0±0 8.0±0 26.0±1.0 14.0±1.0 13.0±1.0 12.0±0 16.0±1.0 9.0±0 10.0±0 10.0±1.0 26.0±1.0
DISC 8.0±0 7.0±0 6.0±0 14.0±1.0 6.0±0 8.0±0 12.0±0 12.0±1.0 8.0±0 8.0±0 8.0±0 21.0±1.0
S. flexneri WELL 11.0±1.0 11.0±1.0 12.0±0 26.0±1.0 11±1.0 10.0±0 15.0±1.0 16.0±1.0 11.0±1.0 10.0±1.0 12.0±1.0 26.0±1.0
DISC 8.0±0 8.0±0 8.0±0 14.0±1.0 8.0±0 8.0±0 13.0±1.0 12.0±1.0 9.0±0 8.0±0 9.0±0 22.0±1.0
B. subtilis WELL 7.0±0 11.0±0 12.0±0 21.0±1.0 7.0±0 8.0±0 6.0±0 21.0±1.0 7.0±0 11.0±1.0 12.0±0 21.0±1.0
DISC 8.0±0 8.0±0 8.0±0 14.0±1.0 9.0±1.0 9.0±0 10.0±1.0 12.0±1.0 9.0±0 9.0±0 8.0±0 12.0±1.0
S. boydii WELL 12.0±1.0 12.0±0 12.0±0 21.0±1.0 14.0±1.0 15.0±1.0 14.0±1.0 26.0±1.0 14.0±0 12.0±0 12.0±0 21.0±1.0
DISC 9.0±1.0 8.0±0 7.0±0 16.0±1.0 8.0±0 8.0±0 8.0±0 12.0±1.0 9.0±0 9.0±0 9.0±0 12.0±1.0
– Nil activity, IL – in vitro leaf, FL – field leaf, CA – callus and C – Control. Contd...
Org name Method Acetone Methanol Aqueous IL FL CA C IL FL CA C IL FL CA C
S. aureus WELL 8.0±0 8.0±0 8.0±0 26.0±1.0 11.0±1.0 12.0±1.0 12.0±1.0 16.0±1.0 12.0±0 10.0±0 10.0±0 23.0±1.0
DISC 10.0±1.0 11.0±1.0 11.0±0 14.0±1.0 12.0±1.0 16.0±1.0 14.0±1.0 16.0±1.0 6.0±0 6.0±0 6.0±0 12.0±1.0
S. typhi WELL 8.0±0 12.0±1.0 12±0 26.0±1.0 12.0±1.0 11.0±1.0 11.0±1.0 21.0±1.0 - - - 23.0±1.0
DISC - - - 25.0±1.0 13.0±1.0 12.0±1.0 11.0±1.0 14.0±1.0 8.0±0 7.0±0 8.0±0 12.0±1.0
C. diphtheriae WELL 11.0±0 12.0±1.0 10.0±0 26.0±1.0 10.0±0 8.0±0 8.0±0 21.0±1.0 11.0±0 12.0±0 11.0±0 23.0±1.0
DISC 8.0±0 9.0±0 10.0±0 12.0±1.0 14.0±1.0 14.0±0 10.0±1.0 15.0±1.0 8.0±0 10.0±0 12.0±0 11.0±1.0
P. vulgaris WELL - 9.0±0 11.0±0 21.0±1.0 11.0±1.0 9.0±0 9.0±0 21.0±1.0 10.0±0 10.0±0 13.0±0 23.0±1.0
DISC - 7.0±0 7.0±0 10.0±1.0 16.0±1.0 16.0±0 12.0±0 16.0±1.0 9.0±0 10.0±0 11.0±0 11.0±1.0
V. vulnificus WELL 7.0±0 7.0±0 9.0±0 21.0±1.0 9.0±0 9.0±0 9.0±0 21.0±1.0 8.0±0 9.0±0 10.0±0 16.0±1.0
DISC 6.0±0 6.0±0 - 15.0±1.0 21.0±0 20.0±0 20.0±0 20.0±1.0 8.0±0 9.0±0 9.0±0 11.0±1.0
S. dysenteriae WELL 10±1.0 9.0±0 8.0±0 21.0±1.0 9.0±0 11.0±0 12.0±0 18.0±1.0 9.0±0 12.0±0 11.0±0 16.0±1.0
DISC 6.0±0 6.0±0 - 12.0±1.0 8.0±0 7.0±0 8.0±0 12.0±1.0 - - - 25.0±1.0
E. coli WELL - - - 21.0±1.0 10.0±0 8.0±0 13.0±0 26.0±1.0 8.0±0 9.0±0 12.0±0 16.0±1.0
DISC 9.0±0 10.0±1.0 10.0±0 12.0±1.0 8.0±0 8.0±0 8.0±0 11.0±1.0 9.0±0 8.0±0 8.0±0 10.0±1.0
E. faecalis WELL 10.0±0 10.0±1.0 10.0±0 26.0±1.0 16.0±0 13.0±0 15.0±0 26.0±1.0 - - - 16.0±1.0
DISC - - - 14.0±1.0 14.0±1.0 17.0±1.0 11.0±0 19.0±1.0 - 9.0±0 10.0±0 10.0±1.0
L. acidophilus WELL 8.0±0 8.0±0 7.0±0 26.0±1.0 10.0±1.0 12.0±0 15.0±0 21.0±1.0 9.0±0 9.0±0 9.0±0 23.0±1.0
DISC 9.0±0 8.0±0 8.0±0 10.0±1.0 13.0±1.0 12.0±0 12.0±0 12.0±1.0 12±0 8.0±0 12.0±0 10.0±1.0
P. aeruginosa WELL 9.0±0 10.0±1.0 11.0±1.0 26.0±1.0 17.0±1.0 16.0±1.0 13.0±1.0 26.0±1.0 8.0±0 12.0±0 11.0±0 23.0±1.0
DISC 6.0±0 6.0±0 6.0±0 14.0±1.0 15.0±1.0 15.0±1.0 13.0±1.0 10.0±1.0 10.0±0 8.0±0 12.0±0 10.0±1.0
S. flexneri WELL 8.0±0 12.0±1.0 11.0±1.0 16.0±1.0 10.0±1.0 9.0±1.0 11.0±1.0 21.0±1.0 - - - 26.0±1.0
DISC 7.0±0 7.0±0 6.0±0 14.0±1.0 9.0±1.0 9.0±1.0 13.0±1.0 20.0±1.0 12.0±1.0 8.0±0 12.0±0 10.0±1.0
B. subtilis WELL 9.0±0 12.0±0 11.0±0 16.0±1.0 14.0±1.0 13.0±1.0 11.0±1.0 21.0±1.0 - - - 21.0±1.0
DISC 9.0±0 7.0±0 7.0±0 14.0±1.0 10.0±0 9.0±0 9.0±0 15.0±1.0 10.0±0 8.0±0 8.0±0 12.0±1.0
S. boydii WELL 8.0±0 8.0±0 8.0±0 16.0±1.0 9.0±0 8.0±0 7.0±0 21.0±1.0 - - - 23.0±1.0
DISC 9.0±0 8.0±0 8.0±0 14.0±1.0 9.0±0 8.0±0 9.0±0 15.0±1.0 8.0±0 8.0±0 10.0±0 12.0±1.0
– Nil activity, IL – in vitro leaf, FL – field leaf, CA – callus and C – Control.
Figure 15a: Antibacterial activity of different solvent extracts of A. javanica (well
diffusion method C – control, IL – in vitro leaf, FL – field leaf, CA – callus),
A. C. diphtheriae, B. P. vulgaris – hexane extract, C. S. dysenteriae – chloroform
extract, D. S. boydii, E. P. aeruginosa – ethyl acetate extract, F. C. diphtheriae –
acetone extract, G. P. aeruginosa, H. E. faecalis – methanol extract, and
I. S. flexneri – aqueous extract.
Figure 15b: Antibacterial activity of different solvent extracts of A. javanica (disc
diffusion method C – control, IL – in vitro leaf, FL – field leaf, CA – callus),
A. S. dysenteriae – chloroform extract, B. P. vulgaris – hexane extract,
C. E. coli – ethyl acetate extract, D. L. acidophilus – acetone extract, E. E. faecalis
- methanol extract, F. P. aeruginosa – aqueous extract. G. S. flexneri,
H. L. acidophilus, and I. S. typhi - methanol extracts
Table 35: Antifungal activity of the extracts of A. javanica
Org name Method Hexane Chloroform Ethyl acetate
IL FL CA C IL FL CA C IL FL CA C
C. albicans WELL 8.1±0 11.0±0 13.3±1.0 16.0±1.0 9.0±0 8.0±0 9.0±0 21.0±1.0 12.0±0 8.0±0 8.0±0 16.0±1.0
DISC - - - 16.0±1.0 - - - 14.0±1.0 - - - 14.0±1.0
A. niger WELL - 11.0±0 12.2±1.0 15.0±1.0 - - - 14.0±1.0 12.0±0 8.0±0 8.0±0 16.0±1.0
DISC - - - 14.0±1.0 - - - 13.0±1.0 9.0±0 9.0±0 8.0±0 14.0±1.0
M. racemosus WELL 9.0±1.0 11.0±1.0 11.4±1.0 16.0±1.0 - - - 16.0±1.0 9.0±0 10.0±0 8.0±0 16.0±1.0
DISC - 9.0±0 - 21.0±1.0 6.0±0 8.0±0 8.0±0 17.0±1.0 8.0±0 9.0±0 - 14.0±1.0
Fusarium sp WELL - - - 14.0±1.0 8.0±0 7.0±0 12.0±1.0 16.0±1.0 - - - 16.0±1.0
DISC 7.0±0 7.0±0 9.2±0 14.0±1.0 9.0±0 7.0±0 9.0±1.0 15.0±1.0 8.0±0 8.0±0 8.0±0 14.0±1.0
Org name Method Acetone Methanol Aqueous
IL FL CA C IL FL CA C IL FL CA C
C. albicans
A.niger
WELL
DISC
WELL
-
-
8.0±0
-
-
8.0±0
-
-
8.0±0
16.0±1.0
16.0±1.0
16.0±1.0
8.0±0
12.0±0
-
8.0±0
13.0±1.0
-
8.0±0
8.0±0
-
14.0±1.0
16.0±1.0
14.0±1.0
8.0±0
-
8.0±0
8.0±0
-
8.0±0
8.0±0
-
8.0±0
26.0±1.0
24.0±1.0
26.0±1.0
DISC - - - 16.0±1.0 - - - 14.0±1.0 9.0±0 9.0±0 8.0±0 24.0±1.0
M. racemosus WELL 8.0±0 9.0±0 - 16.0±1.0 6.0±0 8.0±0 8.0±0 16.0±1.0 9.0±0 10.0±0 8.0±0 26.0±1.0
DISC - 9.0±0 - 16.0±1.0 - - - 16.0±1.0 - - - 24.0±1.0
Fusarium sp WELL - - - 14.0±1.0 8.0±0 7.0±0 12±0 16.0±1.0 - - - 26.0±1.0
DISC - - - 14.0±1.0 9.0±0 7.0±0 9.0±1.0 15.0±1.0 8.0±0 8.0±0 8.0±0 24.0±1.0
– Nil activity, IL – in vitro leaf, FL – field leaf, CA – callus and C – Control.
Figure 16: Antifungal activity of A. javanica well and disc diffusion method,
C – control, IL – in vitro leaf, FL – field leaf, CA – callus, A. C. albicans,
B. Fusarium sp, C. A. niger well diffusion method, D. C. albicans, E. Fusarium sp,
F. A. niger disc diffusion method
Figure 17: Minimum Bactericidal Concentration of A. javanica extract, A – Positive
control, B – in vivo leaf, C – in vitro leaf extract and D – callus extract against
S. aureus.
Table 37: Antibacterial activity of different solvent extracts of A. lanata extracts (in vitro, field leaf and callus)
Org name Method Hexane Chloroform Ethyl acetate
IL FL CA C IL FL CA C IL FL CA C
S. aureus WELL 8.0±0 8.0±0 9.0±0.1 13.0±1.0 9.0±0 8.0±0 8.0.0±0 14.0±1.0 12.0±0 11.0±0 12.0±0 13.0±1.0
DISC 8.0±0 8.0±0 8.0±0 13.0±1.0 8.0±0 8.0±0 8.0±0 13.0±1.0 11.0±0 11.0±0 8.0±0 14.0±1.0
S. typhi WELL 7.0±0 8.0±0 6.0±0 21.0±1.0 7.0±0 8.0±0 6.0±0 11.0±1.0 8.0±0 8.0±0 6.0±0 26.0±1.0
DISC 9.0±0 9.0±0 - 21.0±1.0 9.0±0 9.0±0 9.0±0 13.0±1.0 7.0±0 7.0±0 8.0±0 14.0±1.0
C. diphtheriae WELL 8.0±0 7.0±0 8.0±0 26.0±1.0 11.0±0 11.0±0 11.0±0 26.0±1.0 12.0±0 12.0±0 11.0±0 26.0±1.0
DISC 8.0±0 8.0±0 10.0±0 21.0±1.0 8.0±0 8.0±0 13.0±0 17.0±1.0 7.0±0 9.0±0 8.0±0 14.0±1.0
P. vulgaris WELL 10.0±0 - 11.0±0 26.0±1.0 7.0±0 - - 17.0±1.0 11.0±0 11.0±0 11.0±0 26.0±1.0
DISC 9.0±0 9.0±0 9.0±0 12.0±1.0 8.0±0 8.0±0 9.0±0 15.0±1.0 10.0±0 10.0±0 8.0±0 14.0±1.0
V. vulnificus WELL 10.0±0 10.0±0 - 26.0±1.0 10.0±0 11.0±0 9.0±0 26.0±1.0 10.0±0 7.0±0 - 26.0±1.0
DISC 8.0±0 8.0±0 8.0±0 12.0±1.0 11.0±0 10.0±0 8.0±0 16.0±1.0 12.0±0 14.0±0. - 20.0±1.0
S. dysenteriae WELL 9.0±0 9.0±0 7.0±0 15.0±1.0 8.0±0 6.0±0 8.0±0 15.0±1.0 10.0±0 10.0±0 9.0±0 26.0±1.0
DISC 6.0±0 6.0±0 6.0±0 17.0±1.0 8.0±0 8.0±0 7.0±0 15.0±1.0 7.0±0 7.0±0 9.0±0 12.0±1.0
E. coli WELL 7.0±0 8.0±0 10.0±0 21.0±1.0 8.0±0 8.0±0 10.0±0 21.0±1.0 12.0±0 12.0±0 11.0±0 21.0±1.0
DISC 9.0±0 9.0±0 10.0±0 16.0±1.0 8.0±0 8.0±0 - 15.0±1.0 - - - 16.0±1.0
E. faecalis WELL 6.0±0 6.0±0 - 26.0±1.0 8.0±0 8.0±0 - 26.0±1.0 11.0±0 12.0±0 - 26.0±1.0
DISC 7.0± 12.0±0 10.0±0 12.0±1.0 8.0±0 8.0±0 9.0±0 15.0±1.0 11.0±0 11.0±0 11.0±0 12.0±1.0
L. acidophilus WELL - 11.0±0 7.0±0 18.0±1.0 9.0±0 - 13.0±0 26.0±1.0 10.0±0 8.0±0 8.0±0 16.0±1.0
DISC 9.0±0 8.0±0 8.0±0 11.0±1.0 10.0±0 9.0±0 12.0±0 12.0±1.0 11.0±0 8.0±0 12.0±0 25.0±1.0
P. aeruginosa WELL 8.0±0 8.0±0 10.0±0 26.0±1.0 12.0±0 11.0±0 10.0±0 16.0±1.0 8.0±0 10.0±0 12.0±0 26.0±1.0
DISC 8.0±0 8.0±0 9.0±0 14.0±1.0 9.0±0 9.0±0 10.0±0 12.0±1.0 7.0±0 8.0±0 6.0±0 21.0±1.0
S. flexneri WELL 9.0±0 8.0±0 8.0±0 26.0±1.0 - - 8.0±0 16.0±1.0 8.0±0 - - 26.0±1.0
DISC 12.0±0 12.0±0 11.0±0 14.0±1.0 12.0±0 11.0±0 8.0±0 12.0±1.0 7.0±0 9.0±0 9.0±0 22.0±1.0
B. subtilis WELL 7.0±0 8.0±0 9.0±0 21.0±1.0 10.0±0 9.0±0 11.0±0 21.0±1.0 8.0±0 12.0±0 12.0±0 21.0±1.0
DISC 9.0±0 9.0±0 10.0±0 14.0±1.0 12.0±0 12.0±0 11.0±0 12.0±1.0 8.0±0 8.0±0 7.0±0 12.0±1.0
S. boydii WELL 9.0±0 10.0±0 11.0±0 21.0±1.0 10.0±0 8.0±0 10.0±0 26.0±1.0 15.0±0 10.0±0 12.0±0 21.0±1.0
DISC 10.0±0 9.0±0 11.0±0 16.0±1.0 8.0±0 9.0±0 10.0±0 12.0±1.0 10.0±0 10.0±0 13.0±0 12.0±1.0
– Nil activity, IL – in vitro leaf, FL – field leaf, CA – callus and C – Control. Contd…
Org name Method Acetone
IL FL
CA
C
Methanol
IL FL
CA
C
Aqueous
IL FL
CA
C
S. aureus WELL 10.0±0 10.0±0 9.0±0 26.0±1.0 12.0±0 11.0±0 14.0±0 16.0±1.0 - - - 23.0±1.0
DISC 10.0±0 8.0±0 9.0±0 14.0±1.0 7.0±0 7.0±0 6.0±0 16.0±1.0 8.0±0 8.0±0 7.0±0 12.0±1.0
S. typhi WELL 7.0±0 7.0±0 6.0±0 26.0±1.0 12.0±0 12.0±0 11.0±0 21.0±1.0 7.0±0 7.0±0 7.0±0 23.0±1.0
DISC 9.0±0 9.0±0 9.0±0 25.0±1.0 11.0±0 11.0±0 11.0±0 14.0±1.0 6.0±0 7.0±0 8.0±0 12.0±1.0
C. diphtheriae WELL 7.0±0 7.0±0 8.0±0 26.0±1.0 7.0±0 7.0±0 8.0±0 21.0±1.0 8.0±0 8.0±0 11.0±0 23.0±1.0
DISC 8.0±0 8.0±0 7.0±0 12.0±1.0 9.0±0 9.0±0 8.0±0 10.0±1.0 12.0±0 12.0±0 10.0±0 11.0±1.0
P. vulgaris WELL 11.0±0 - 8.0±0 11.0±1.0 16.0±0 14.0±0 12.0±0 21.0±1.0 8.0±0 - 8.0±0 23.0±1.0
DISC 11.0±0 10.0±0 - 10.0±1.0 10.0±0 10.0±0 10.0±0 16.0±1.0 12.0±0 10.0±0 11.0±0 11.0±1.0
V. vulnificus WELL 8.0±0 11.0±0 9.0±0 21.0±1.0 - 8.0±0 9.0±0 21.0±1.0 16.0±0 13.0±0 10.0±0 16.0±1.0
DISC - - - 15.0±1.0 - - - 20.0±1.0 - - - 11.0±1.0
S. dysenteriae WELL 9.0±0 - 8.0±0 21.0±1.0 12.0±0 8.0±0 12.0±0 18.0±1.0 - - 11.0±0 16.0±1.0
DISC 8.0±0 8.0±0 7.0±0 21.0±1.0 9.0±0 9.0±0 8.0±0 12.0±1.0 12.0±0 12.0±0 11.0±0 25.0±1.0
E. coli WELL 8.0±0 8.0±0 11.0±0 21.0±1.0 7.0±0 7.0±0 8.0±0 26.0±1.0 6.0±0 6.0±0 7.0±0 16.0±1.0
DISC 9.0±0 9.0±0 11.0±0 12.0±1.0 11.0±0 11.0±0 9.0±0 11.0±1.0 12.0±0 11.0±0 9.0±0 10.0±1.0
E. faecalis WELL 6.0±0 6.0±0 - 26.0±1.0 14.0±0 16.0±0 - 26.0±1.0 6.0±0 6.0±0 6.0±0 16.0±1.0
DISC - - - 14.0±1.0 - - - 19.0±1.0 - - - 10.0±1.0
L. acidophilus WELL 13.0±0 11.0±0 12.0±0 26.0±1.0 16.0±0 15.0±0 18.0±0 21.0±1.0 10.0±0 10.0±0 10.0±0 23.0±1.0
DISC 10.0±0 12.0±0 11.0±0 10.0±1.0 18.0±0 11.0±0 8.0±0 12.0±1.0 8.0±0 8.0±0 8.0±0 10.0±1.0
P. aeruginosa WELL 8.0±0 8.0±0 13.0±0 26.0±1.0 8.0±0 8.0±0 8.0±0 26.0±1.0 10.0±0 11.0±0 11.0±0 23.0±1.0
DISC 9.0±0 9.0±0 9.0±0 14.0±1.0 8.0±0 9.0±0 8.0±0 10.0±1.0 7.0±0 8.0±0 8.0±0 10.0±1.0
S. flexneri WELL - - 11.0±0 16.0±1.0 - - - 21.0±10 - - - 26.0±1.0
DISC 13.0±0 13.0±0 12.0±0 16.0±1.0 14.0±0 13.0±0 12.0±0 20.0±1.0 13.0±0 14.0±0 10.0±0 10.0±1.0
B. subtilis WELL - 6.0±0 8.0±0 16.0±1.0 - - 8.0±0 21.0±1.0 8.0±0 8.0±0 11.0±0 21.0±1.0
DISC 12.0±0 8.0±0 8.0±0 14.0±1.0 8.0±0 9.0±0 9.0±0 15.0±1.0 12.0±0 9.0±0 9.0±0 12.0±1.0
S. boydii WELL 6.0±0 6.0±0 8.0±0 16.0±1.0 8.0±0 8.0±0 7.0±0 21.0±1.0 9.0±0 7.0±0 7.0±0 23.0±1.0
DISC 11.0±0 11.0±0 12.0±0 14.0±1.0 12.0±0 11.0±0 11.0±0 15.0±1.0 10.0±0 9.0±0 9.0±0 12.0±1.0
– Nil activity, IL – in vitro leaf, FL – field leaf, CA – callus and C – Control.
Figure 18a: Antibacterial activity of A. lanata (well diffusion method C – control,
IL – in vitro leaf, FL – field leaf, CA – callus), A. C. diphtheriae, B. S. flexneri,
C. L. acidophilus – methanol extract, D. S. boydii, E. S. typhi, F. S. dysenteriae –
ethyl acetate extract, G. B. subtilis, H. S. aureus, I. S. dysenteriae – acetone extract
Figure 18b: Antibacterial activity of A. lanata (disc diffusion method C – control,
IL – in vitro leaf, FL – field leaf and CA – callus), A. S. flexneri, B. L. acidophilus,
C. S. typhi – methanol extract, D. S. aureus, E. S. dysenteriae, F. B. subtilis –
acetone extract, G. S. typhi – ethyl acetate extract, H. C. diphtheriae – aqueous
extract, I. P. aeruginosa – methanol extract.
Table 38: Antifungal activity of the extracts of A. lanata
Org name Method Hexane Chloroform Ethyl acetate
IL FL CA C IL FL CA C IL FL CA C
C. albicans WELL 8.0±0 8.0±0 8.0±0 21.0±1.0 8±1.0 8.0±0 8±1.0 21.0±1.0 9.0±0 9.0±0 9.0±0 26.0±1.0
DISC 9.0±0 9.0±0 9.0±0 10.0±1.0 9.0±0 9.0±0 9.0±0 10.0±1.0 8.0±0 9.0±0 9.0±0 14.0±1.0
A. niger WELL 10.0±0 11±0 11.0±0 21.5±1.0 12.0±0 11.0±0 11.0±0 15.0±1.0 8.0±0 8.0±0 8.0±0 26.0±1.0
DISC 9.0±0 9.0±0 9.0±0 21.0±1.0 9.0±1.
0
9±1.0 9±1.0 13.0±1.0 9.0±0 9.0±0 9.0±0 14.0±1.0
M.
racemosus
WELL - - - 26.0±1.0 - - - 26.0±1.0 - - - 26.0±1.0
DISC - - - 21.0±1.0 - - - 17.0±1.0 - - - 14.0±1.0
Fusarium
sp.
WELL - - - 26.0±1.0 - - - 10.0±1.0 - - - 26.0±1.0
DISC - - - 12.0±1.0 - - - 15.0±1.0 - - - 14.0±1.0
Org name Method Acetone Methanol Aqueous
IL FL CA C IL FL CA C IL FL CA C
C. albicans WELL 8.0±0 8.0±0 - 21.0±1.0 8.0±0 8.0±0 12.0±0 21.0±1.0 - - - 26.0±1.0
DISC 11.0±0 10.0±0 10.0±0 10.0±1.0 10±1.0 10.0±0 10±1.0 10.0±1.0 9.0±0 9.0±0 10.0±0 14.0±1.0
A. niger WELL - - - 21.0±1.0 9±0.1 9.0±0 9.0±0 15.0±1.0 12.0±0 8.0±0 8.0±0 26.0±1.0
DISC 8.0±0 8.0±0 8.0±0 21.0±1.0 7±0.1 7.0±0 7.0±0 13.0±1.0 8.0±0 8.0±0 8.0±0 14.0±1.0
M.
racemosus
WELL 8.0±0 7.0±0 7.0±0 26.0±1.0 8±1.0 9.0±0 7.0±0 26.0±1.0 8.0±0 7.0±0 7.0±0 26.0±1.0
DISC - - - 21.0±1.0 - - - 17.0±1.0 - - - 14.0±1.0
Fusarium
sp.
WELL 8.0±0 8.0±0 7.0±0 26.0±1.0 9±1.0 9.0±0 9.0±0 16.0±1.0 9.0±0 8.0±0 8.0±0 26.0±1.0
DISC - - - 12.0±1.0 - - - 15.0±1.0 - - - 14.0±1.0
– Nil activity, IL – in vitro leaf, FL – field leaf, CA – callus and C – Control.
Figure 19: Antifungal activity of A. lanata (C – control, IL – in vitro leaf, FL – field
leaf, CA – callus) A. A. niger, B. Fusarium sp, C. M. racemosus well diffusion
method D. A. niger, E. Fusarium sp, F. C. albicans disc diffusion method
Figure 20: Minimum Bactericidal Concentration of A. lanata extracts, A – Positive
control, B – in vivo leaf, C – in vitro leaf extract and D – callus extract against E.coli
Figure 21: Bioautography of TLC fractions against S. aureus & S. typhi, A – AJ IL
F1, B – AJ FL F1, C – AJ CA F1 against S. aureus, D – AJ FL F1, E – AJ IL F1,
and F – AJ CA F1 against S. typhi
Figure 22: Bioautography of TLC fractions of A. lanata, A – AL FL F1, B – AL IL
F1, C – AL CA F1 (S. aureus), and D – AL CA F1 (S. typhi)
Figure 23: Preliminary phytochemical of A. javanica extracts
Table 40: Preliminary phytochemical analysis of extracts of A. javanica
HX CL EA AC MT AQ
Tests FL IL CA FL IL CA FL IL CA FL IL CA FL IL CA FL IL CA
Alkaloids + + - + + - + + + + + + - + - + + +
Flavonoids + + + + + + + + + + + + + + + + + +
Phenolic + + + + + + + + + + + + + + + + + +
Terpenoids + + - - - - - - - - - - - - - - - -
Tannins + + + + + + + + + + + + + + + + + +
Saponins + + + + + - - - - + + + + + + + + +
Carbohydrates + + + + + + + + + + + + + + + + + +
Amino acids - - - - - - + + - + - - + + + + - -
Glycosides + + - + + + + + - + + - + + + + + +
Fixed oils - - - - - - - - - + + + + + + + - -
` HX – hexane; CL – chloroform; EA – ethyl acetate; AC – acetone; MT – methanol; AQ – aqueous; FL – in vivo leaf; IL – in vitro
leaf; CA – callus; + = indicates presence; – = indicates absence.
Table 41: Quantification of phytochemicals (total phenols, tannins) from different extracts of A. javanica
Total phenolic(mg GAE/g) Tannins (mg GAE/g)
Solvents In vivo leaf In vitro leaf Callus In vivo leaf In vitro leaf Callus
Hexane 105.64 ± 1.79c 115.32 ± 14.21
c 83.44 ± 2.40
b 15.7 ± 4.11
f 16.32 ± 6.30
f 8.23 ± 1.37
f
Chloroform 54.19 ± 1.98d 41.21 ± 0.50
e 61.44 ± 4.05
c 29.03 ± 6.69
e 31.98 ± 4.31
e 4.9 ± 1.57
f
Ethyl acetate 30.21 ± 0.82e 65.39 ± 4.64
d 55.24 ± 1.32
c 36.17 ± 4.74
e 30.63 ± 2.86
e 10.51 ± 0.76
f
Acetone 33.24 ± 2.91e 73.95 ± 7.90
d 111.54 ± 6.93
c 34.37 ± 5.98
e 31.38 ± 4.09
e 13.65 ± 4.05
f
Methanol*
150.25 ± 5.29a 155.51 ± 4.72
a 145.23 ± 1.68
a 82.01 ± 2.76
b 87.60 ± 4.24
b 30.84 ± 4.9
a
Aqueous*
156.68 ± 1.45a 139.20 ± 5.44
a 124.65 ± 4.16
a 76.08 ± 3.71
b 78.23 ± 7.02
b 34.71 ± 5.7
a
GAE – Gallic acid equivalents, values in the table are the mean ± SE of triplicates, Superscript in the each row carrying different letters
are significantly different at P<0.05 or 0.01, *-Significance at 0.05, by Newman-Keuls Multiple Comparison Test
Table 42: Quantification of phytochemicals (flavonoids and carbohydrates) from different extracts of A. javanica
Flavonoids (mg RE/g) CHO (mg GE/g)
Solvents In vivo leaf In vitro leaf Callus In vivo leaf In vitro leaf Callus
Hexane 61.59 ± 0.7d 30.58 ± 1.8
e 41.01 ± 2.2
e 126.95 ± 4.4
a 110.54 ± 2.3
a 88.66 ± 3.7
b
Chloroform 57.25 ± 6.8c 44.49 ± 4.9
e 40.14 ± 5.3
e 120.00 ± 3.4
a 113.59 ± 4.3
a 75.16 ± 4.8
b
Ethyl acetate 88.84 ± 2.2b 93.77 ± 2.4
b 83.62 ± 6.4
b 116.87 ± 0.6
a 116.29 ± 5.0
a 80.15 ± 4.6
b
Acetone*
105.94 ± 2.3a 67.97 ± 7.2
d 145.65 ± 0.9
ab 117.23 ± 3.0
a 121.43 ± 0.3
a 88.99 ± 3.6
b
Methanol#
141.01 ± 2.1ab
153.19 ± 2.0abc
177.54 ± 1.8abc
139.00 ± 2.1ab
127.28 ± 4.3a 107.72 ± 1.6
a
Aqueous*
109.42 ± 3.6a 104.49 ± 3.8
a 104.20 ± 2.0
a 127.77 ± 1.1
a 114.12 ± 1.3
a 102.85 ± 1.2
a
RE – Rutin equivalents, GE – glucose equivalents, values in the table are the mean ± SE of triplicates, Superscript in the each row
carrying different letters are significantly different at P<0.05 or 0.01, *-Significance at 0.05, # - Significance at 0.01 by Newman-Keuls
Multiple Comparison Test
Figure 24: Preliminary phytochemicals of A. lanata extracts
HX CL EA AC MT AQ FL IL CA FL IL CA FL IL CA FL IL CA FL IL CA FL IL CA
+ + - + + - + + - + + - + + + + + -
+ + + + + + + + + + + + + + + + + +
+ + + + + + + + + + + + + + + + + +
- + - - - - + - - + + + - - - - - -
+ + + + + + + + + + + + + + + + + +
- - - - - - - - - + + - + + + + - -
+ + + + + + + + + + + + + + + + + +
+ - - + - - + + - + + - + + - + + -
+ - - + - - - - - + + - + + + + + +
- - - - - - - - - - - - - - - - - -
Table 43: Preliminary Phytochemical analysis of A. lanata extracts
Tests
Alkaloids
Flavonoids
Phenolic
Terpenoids
Tannins
Saponins
Carbohydrates
Amino acids
Glycosides
Fixed oils
HX – hexane; CL – chloroform; EA – ethyl acetate; AC – acetone; MT – methanol; AQ – aqueous; FL – in vivo leaf; IL – in vitro leaf;
CA – callus; + = indicates presence; – = indicates absence.
Table 44: Quantification of phytochemicals (total phenols, tannins) from different extracts of A. lanata
Total phenolic (mg GAE/g extract) Tannins (mg GAE/g extract)
Solvents Field leaf In vitro leaf Callus Field Leaf In vitro leaf Callus
Hexane 23.23 ± 1.14d 27.91 ± 2.14
d 24.81 ± 1.66
d 8.71 ± 1.39
d 20.32 ± 4.31
d 8.23 ± 1.37
d
Chloroform 20.00 ± 1.89 d 34.10 ± 2.25
d 17.76 ± 0.69
d 5.69 ± 2.33
d 15.31 ± 2.77
d 4.90 ± 1.57
d
Ethyl acetate 28.23 ± 0.82 d 55.18 ± 1.37
b 15.72 ± 1.32
d 9.50 ± 3.21
d 13.30 ± 4.57
d 10.51 ± 0.76
d
Acetone 28.63 ± 2.50 d 54.19 ± 2.90
b 34.62 ± 2.24
e 11.04 ± 1.61
d 26.71 ± 3.42
d 13.65 ± 4.05
d
Methanol#
193.98 ± 0.44abc
137.85 ± 2.44abc
69.37 ± 3.59b 109.41 ± 3.59
abc 82.60 ± 2.24
b 36.84 ± 2.12
b
Aqueous*
120.72 ± 0.58abc
137.65 ± 2.60abc
60.67 ± 2.97b 87.01 ± 1.42
b 78.23 ± 2.02
b 36.05 ± 2.13
b
GAE – gallic acid equivalents, values in the table are the mean ± SE of triplicates, Superscript in the each row carrying different letters
are significantly different at P<0.05 or 0.01, *-Significance at 0.05, # - Significance at 0.01 by Newman-Keuls Multiple Comparison
Test
Table 45: Quantification of phytochemicals (flavonoids and carbohydrates) from different extracts of A. lanata
Flavonoids (mg RE/g) Carbohydrates (mg GE/g)
Solvents In vivo Leaf In vitro leaf Callus in vivo leaf In vitro leaf Callus
Hexane 29.71 ± 3.05c 21.88 ± 1.81
c 23.62 ± 2.19
c 31.07 ± 4.8
b 14.73 ± 6.3
b 5.70 ± 0.6
d
Chloroform 28.26 ± 2.30c 23.91 ± 0.87
c 25.65 ± 1.74
c 36.14 ± 4.1
c 9.66 ± 2.9
c 6.03 ± 0.1
c
Ethyl acetate 54.06 ± 3.53b 35.22 ± 2.30
c 41.30 ± 3.14
b 26.92 ± 0.4
c 15.65 ± 4.4
c 5.18 ± 0.2
c
Acetone*
92.03 ± 2.53a 88.26 ± 3.48
b 150.58 ± 4.29
abc 27.31 ± 0.5
c 14.66 ± 4.9
c 6.10 ± 0.2
c
Methanol#
135.80 ± 4.46ab
118.41 ± 3.92ab
151.45 ± 4.97abc
46.28 ± 3.6b 58.60 ± 3.2
ab 9.57 ± 0.5
c
Aqueous*
110.00 ± 1.74ab
96.38 ± 4.10ab
90.00 ± 2.70ab
20.92 ± 0.3c 17.96 ± 3.7
c 7.09 ± 1.0
c
RE – rutin equivalents, GE – Glucose equivalents, values in the table are the mean ± SE of triplicates, Superscript in the each row
carrying different letters are significantly different at P<0.05 or 0.01, *-Significance at 0.05, # - Significance at 0.01 by Newman-Keuls
Multiple Comparison Test
Table 47: FRAP, phosphomolybdenum and metal chelation assay of A. javanica extracts
Solvents
FRAP (mM (Fe)II/g extract) Phosphomolybdenum (mg AAE/g) Metal chelation (mg EDTAE/g)
In vivo leaf In vitro leaf Callus In vivo leaf In vitro leaf Callus In vivo leaf In vitro leaf Callus
Hexane 43.90 ± 1.9b
15.22 ± 1.0c
10.66 ± 6.4c
86.95 ± 2.4c
100.54 ± 2.3bc
78.66 ± 3.7c
89.32 ± 3.0c
93.16 ± 3.7b
98.38 ± 1.5bc
Chloroform 23.61 ± 4.6d
09.20 ± 0.7c
17.97 ± 0.7c
90.00 ± 3.4b
103.59 ± 2.3bc
85.16 ± 2.8c
110.36 ± 4.4bc
118.81 ± 2.0bc
95.35 ± 1.6b
Ethyl acetate 27.58 ± 4.1c
16.72 ± 1.7c
14.70 ± 0.5c
76.87 ± 0.6c
106.29 ± 2.0bc
83.15 ± 4.6c
133.70 ± 0.8a
112.69 ± 4.9bc
99.39 ± 0.5bc
Acetone 20.93 ± 2.7c
33.22 ± 3.3b
15.92 ± 0.4c
217.23 ± 2.0ab
121.43 ± 0.3b
87.99 ± 3.6c
167.27 ± 3.6a
43.27 ± 1.2c
97.71 ± 0.5b
Methanol* 124.26 ± 1.7a 157.01 ± 2.7a 130.18 ± 0.1a 379.00 ± 1.1a 377.28 ± 2.3a 167.72 ± 1.6b 178.96 ± 1.8a 126.75 ± 5.3b 98.63 ± 2.2bc
Aqueous 92.00 ± 6.7b
71.39 ± 0.3b
78.53 ± 2.2a
227.77 ± 1.1ab
344.12 ± 3.3a
142.85 ± 1.2b
127.22 ± 1.1ab
114.18 ± 1.0bc
17.32 ± 2.0c
AAE – ascorbic acid equivalents, EDTAE – ethylene diamine tetra acetic acid equivalents, values in the table are the mean ± SE of
triplicates, Superscript in the each row carrying different letters are significantly different at P<0.05 *-Significance at 0.05, Newman-
Keuls Multiple Comparison Test
Table 49: Total antioxidant (FRAP, phosphomolybdenum, metal chelation) potential of the extracts of the A. lanata extracts
Samples
FRAP (mM (Fe)II/g) Metal chelation (mg EDTAE/g) Phosphomolybdenum (mg AAE/g)
In vivo Leaf In vitro leaf Callus In vivo Leaf In vitro leaf Callus In vivo Leaf In vitro leaf Callus
Hexane 5.58 ± 4.04c
11.74 ± 4.15c
3.70 ± 0.47c
87.10 ± 1.01c
8.73 ± 3.88c
14.21 ± 1.52c
40.89 ± 3.11cd
33.18 ± 5.25cd
26.85 ± 3.69c
Chloroform 9.69 ± 2.06c 11.29 ± 3.41c 10.32 ± 4.10c 112.69 ± 6.36b 34.63 ± 7.04c 11.18 ± 1.59c 107.27± 3.40bc 74.80 ± 4.02c 77.10 ± 4.01c
Ethyl acetate 5.10 ± 0.22c
7.74 ± 1.76c
4.88 ± 0.58c
193.27 ±1.05ab
28.52 ± 4.93c
15.22 ± 0.54c
80.90 ± 2.22c
94.78 ± 5.00c
42.85 ± 2.40cd
Acetone 3.53 ± 0.30c
28.00 ± 1.89c
13.38 ± 1.91c
186.85 ± 4.41ab
127.45 ± 1.18b
13.53 ± 0.46c
356.95 ± 0.78a
111.07 ± 5.68bc
70.71 ± 3.64c
Methanol# 112.88 ±2.80a 73.48 ± 2.75b 33.29 ± 0.12b 282.19 ± 6.35a 242.57 ± 5.28a 114.46 ± 2.15a 400.48 ± 4.10a 345.90 ± 4.32a 102.55 ± 0.80b
Aqueous*
64.02 ± 3.82b
67.22 ± 3.30b
25.77 ± 0.78b
186.68 ± 3.37ab
185.56 ± 1.01ab
94.02 ± 3.95ab
206.29 ± 1.12b
204.57 ± 6.74b
91.50 ± 3.20c
AAE – ascorbic acid equivalents, EDTAE – ethylene diamine tetra acetic acid equivalents, values in the table are the mean ± SE of
triplicates, Superscript in the each row carrying different letters are significantly different at P<0.05 or 0.01, *-Significance at 0.05, # -
Significance at 0.01 by Newman-Keuls Multiple Comparison Test
Figure 25a: Different concentrations of A. javanica in vivo leaf extract in MTT assay
Figure 25b: Different concentrations of A. javanica in vitro leaf extract
Figure 25c: Different concentrations of A. javanica callus leaf extract
Figure 26: DNA fragmentation of MCF – 7 cell line, M – Marker, L1 Control, L2 -
in vivo sample, L3 - in vitro sample and L4 - callus sample
Figure 27a: Different concentrations of A. lanata in vivo leaf extract
Figure 27b: Different concentrations of A. lanata in vitro leaf extract
Figure 27c: Different concentrations of A. lanata callus leaf extract
Figure 28: DNA fragmentation of MCF – 7 cell lines by A. Lanata, M – Marker,
C – control, L1 – in vivo, L2 – in vitro, L3 – callus (IC50 concentration).
Table 52: Rf values of the compounds separated from A. javanica extracts
Extract No. of Rf value Interaction Fractions
AJ – IL – MT 02 0.888, 0.944 polyphenols and polysaccharides (Martin, et.al, 1986)
AJ – FL – MT 03 0.866, 0.877, 0.9 polyphenols and proteins, (Bate-Smith, 1981)
AJ – CA – MT 01 0.877 Polyphenol (Bate-Smith, 1981)
AJ – IL – MT = A. javanica in vitro leaf Methanol extract, AJ – FL – MT = A. javanica
in vivo leaf Methanol extract, AJ – CA – MT = A. javanica callus Methanol extract
Table 53: Rf values of the compounds separated from A. lanata extracts
Extract No. of Rf value Interaction Fractions
AL – IL – MT 03 0.800, 0.811, 0.854 polyphenols (Martin, et.al, 1986)
AL – FL – MT 04 0.245, 0.249, 0.414,
0.870,
Fatty acids, amino acids, and polyphenols (Bate-
Smith, 1981)
AL – CA – MT 02 0.214, 0.865 polysaccharides and Polyphenol (Bate-Smith,
1981)
AL – IL – MT = A. lanata in vitro leaf Methanol extract, AL – FL – MT = A. lanata in
vivo leaf Methanol extract, AL – CA – MT = A. lanata callus Methanol extract
Figure 29a: TLC fraction of A. javanica extracts, A – In vivo leaf, B – In vitro leaf,
C – Callus extract
Figure 29b: TLC fraction of A. lanata extracts, A – In vivo leaf, B – In vitro leaf, C
– Callus extract
Figure 30a: HPLC chromatogram of AJ-FL-F1 (A. javanica field leaf fraction 1)
Figure 30b: HPLC chromatogram of AJ-IL-F1 (A. javanica in vitro leaf fraction 1)
Figure 30c: HPLC chromatogram of AJ-CA-F1 (A. javanica callus fraction 1)
Figure 30d: HPLC chromatogram of standards 1 – Gallic acid, 2 – Catechin, 3 – Epicatechin, 4 – Quercetin
Figure 31a: HPLC chromatogram of AL– FL– F1 (A. lanata field leaf fraction 1)
Figure 31b: HPLC chromatogram of AL–IL–F1 (A. lanata in vitro leaf fraction 1)
Figure 31c: HPLC chromatogram of AL–CA–F1 (A. lanata callus fraction 1)
Figure 31d: HPLC chromatogram of standards 1 – Chlorogenic acid, 2 – Vanillin, 3 – Ferulic acid, 4 – Resveratrol
Table 56: Bioactive compounds detected in A. javanica extracts by using GC – MS
Plant Sample RT Compound MW Formula Reported property Author
A. In vivo 15.73 MYO-INOSITOL, 4-C-METHYL- 194 C7H14O6 Antitumor Hudlicky et al., 2002
javanica leaf 28.704 STIGMASTEROL 83-48-7 412 C29H48O Antioxidant, anti-inflammatory,
immunomodulatory
Hassanein et al, 2012,
Norman et al., 1972
29.359 3BETA-HYDROXY-5-CHOLEN-24-
OIC ACID 5255-17-4
374 C24H38O3 Antibacterial Batte et al., 1990
30.37
HENTRIACONTANE
436
C31H64
Antimicrobial
Milen georgiev et al.,
2010
In vitro
15.839
MYO-INOSITOL, 4-C-METHYL-
194
C7H14O6
Antitumor
Hudlicky et al., 20002
leaf 16.424 .BETA.-D-GLUCOPYRANOSIDE,
METHYL
194 C7H14O6 Antiallergic, Cytotoxic,
antidiabetic
Kim et al., 2000, Lee &
Sohn 2008
28.679 STIGMASTEROL 412 C29H48O Antioxidant, anti-inflammatory,
immunomodulatory
Hassanein et al, 2012,
Norman et al., 1972
Callus 15.34 .BETA.-D-GLUCOPYRANOSIDE,
METHYL
194 C7H14O6 Antiallergic, Cytotoxic,
antidiabetic
Kim et al., 2000, Lee &
Sohn 2008
19.810 Olyl alcohol 268 C18H36O Antifungal Dawn et al., 1962
20.025 Olic acid 282 C18H34O2 Antioxidant Visioli & Galli 2002
28.67 STIGMASTEROL 83-48-7 412 C29H48O Antioxidant, anti-inflammatory,
immunomodulatory
Hassanein et al, 2012,
Norman et al., 1972,
Figure 32a: GC – MS chromatogram of A. javanica in vivo leaf methanol extract
Figure 32b: GC – MS chromatogram of A. javanica in vitro leaf methanol extract
Figure 32c: GC – MS chromatogram of A. javanica callus methanol extract
Table 57: Bioactive compounds detected in A. lanata extracts by using GC – MS
Plant Sample RT Compound MW Formula Reported property Author
A. In vivo 17.85 METHYL 11-METHYL-DODECANOATE 228 C14H28O2 Antibacterial Ntougias et al., 2000
lanata leaf 20.32
3-METHYL-2-(2-OXOPROPYL)FURAN
138
C8H10O2
Anti-inflammatory
Berger et al., 2002
30.37
HENTRIACONTANE
436 C31H64
Antimicrobial
Milen georgiev et al., 2010
In vitro
29.37
10,12,14-NONACOSATRIYNOIC ACID
426 C29H46O2
Antioxidant
Yang et al., 2011
leaf 16.85
Z,Z-6,28-HEPTATRIACTONTADIEN-2-ONE
530
C37H70O
Antimicrobial
Milengeorgiev et al., 2010
23.11
1,2-BENZENEDICARBOXYLIC ACID,
278
C16H22O4
Antianderogenic property,
Koch et al., 2003,
MONO(2-ETHYLHEXYL) ESTER Antidiabetic Maruthupandian and Mohan,
2011
Callus
16.94
3,7,11,15-TETRAMETHYL-2-HEXADECEN-1-
296
C20H40O
Antibacterial, antifungal
Ogunlein et al., 2009
OL
23.54
PHENOL, 3,5-BIS(1,1-DIMETHYLETHYL)-
206
C14H22O
Antiinflammatory
Unangst et al., 1992
25.05
CYCLOTRISILOXANE, HEXAMETHYL-
222
C6H18O3Si3
Biohealth function
Xie and Li 2010
Figure 33a: GC – MS chromatogram of A. lanata methanol in vivo leaf extract
Figure 33b: GC – MS chromatogram of A. lanata methanol in vitro leaf extract
Figure 33c: GC – MS chromatogram of A. lanata methanol callus leaf extract
Table 58: FT – IR analysis of A. javanica samples Vibrations
O-H stretch.
N-H stretch (amines)
cm-1
AJ-FL-MT AJ –FL-F1 AJ-IL-MT AJ-IL-F1 AJ-CA-MT AJ-CAF1
3396 3411 3371 3395 3396 3415
C-H stretch. 2946 2948 2946 2947 2943 2969
-O-CH3 asym. stretch.
C-H stretch (aldehydes)
2834 2838 2834 2839 2828 2842
S-H stretch. 2531 2524 2528 2523 2522 2523
C=C stretch.
C=N stretch
N-H defor.
O=C-O asym. stretch.
NO2 stretch.
C=C stretch.
C=N stretch.
-N=N stretch.
C=O stretch. (phenols, ketones, aldehydes, carboxylic
acids, carboxylic ester, lactones,amides)
C-H defor.
O-H bending in plane
C=O stretch.
2048 2048 2046 2074 2053 2075 1567 1649 1570 1648 1571 1647 1404 1412 1404 1414 1406 1412
Contd...
R-O-R stretch.
C-OH stretch.
C-H stretch. (amine)
1118 1112 1117 1112 1118 1112
C-OH stretch.
P-O strerch.
C-OC asym. Stretch (ether)
1027 1052 1026 1053 1027 1031
O-H deform.
C-H stretch.
- - - - 953 1015
C-N vib.
N-O stretch.
C-H bending (out of plane). (aromatic hydrocarbones)
- - - - 829 -
O-H bending (out of plane)
C-O stretch.
C-N stretch.
C-H bending (out of plane), (aromatic hydrocarbones)
751 - 740 - 752 -
C-H bending. (alkynes)
N-H bending out of plane (amide, lactone)
696 - 696 - - -
C-H bending (out of plane). (aromatic hydrocarbones) - - - - 702 691
O-N=O defor. 652 660 653 659 653 -
AJ – IL – MT = A. javanica in vitro leaf Methanol extract, AJ – FL – MT = A. javanica in vivo leaf Methanol extract, AJ – CA – MT =
A. javanica callus Methanol extract; AJFLF1 – A. javanica field leaf fraction 1, AJILF1 – A. javanica in vitro leaf fraction 1, AJCAF1 –
A. javanica callus fraction 1.
Figure 34a: FT – IR spectrum for crude sample of A. javanica (AJ MT FL – A. javanica methanol field leaf)
Figure 34b: FT – IR spectrum for fraction of A. javanica (AJ FL F1 – A. javanica field leaf fraction 1)
Figure 35a: FT – IR spectrum for crude sample of A. javanica (AJ MT IL A. javanica methanol in vitro leaf)
Figure 35b: FT – IR spectrum for fraction of A. javanica (AJ IL F1 – A. javanica in vitro leaf fraction 1)
Figure 36a: FT – IR spectrum for crude sample of A. javanica (AJ MT CA A. javanica methanol callus)
Figure 36b: FT – IR spectrum for fraction of A. javanica (AJ CA F1 – A. javanica callus fraction 1)
Table 59: FT – IR analysis of A. lanata samples
Vibrations cm-1
AL FL MT AL FL F1 AL IL MT AL IL F1 AL CA MT AL CA F1
O-H3 Trans stretch (amines) 3433 3372 3370 3368 3408 3412
C-H stretch. 2923 2946 2947 2946 2922 2948
-O-CH3 asym 2858 2866 - 2867 - 2836
C-H stretch (aldehydes) 2360 2221 2213 2221 2360 -
C=C stretch, C=O stretch 2098 2046 - 2047 - 2074
NO2 stretch, O=C-O asym 1717 - - - 1717 -
O-H bending in plane, C=O stretch. (Ketones, aldehydes, carboxylic acids, carboxylic ester, lactones,amides)
1629 1659 1571 1662 1616 1650
C-H defor. O-H bending in plane C=O stretch.
1455 1453 1477 1453 1457 1454
C-H stretch. (amine) 1368 1415 1403 1414 1373 1411
R-O-R stretch. 1274 1113 - 1113 1219 1113
C-OH stretch. 1219 - - - - -
P-O stretch 1088 - 1120 - 1092 -
S=O Str - 1027 1026 1028 - 1026
C-H def (benzene ring with 1 free H) - - 927 - - -
C-H def (Benzene ring) - - 830 - - -
C-H bending (out of plane), (aromatic hydrocarbones) 770 - 753 - 771 -
C-H def (R1CH=CHR2) - - 696 - - 690
N-H bending out of plane (amide, lactone) 663 660 651 658 666 -
C-H - - 620 - - -
O-N=O defor. 598 - - - 595 -
O-N stretch 459 - - - 457 -
AL – IL – MT = A. lanata in vitro leaf Methanol extract, AL – FL – MT = A. lanata in vivo leaf Methanol extract, AL – CA – MT = A. lanata
callus Methanol extract, ALFLF1 – A. lanata field leaf fraction 1, ALILF1 – A. lanata in vitro leaf fraction 1, ALCAF1 – A. lanata callus fraction 1.
Figure 37a: FT – IR spectrum for crude sample of A. lanata (AL MT FL – A. lanata methanol field leaf)
Figure 37b: FT – IR spectrum for fraction of A. lanata (AL FL F1 – A. lanata field leaf fraction 1)
Figure 38a: FT – IR spectrum for crude sample of A. lanata (AL MT IL – A. lanata methanol in vitro leaf)
Figure 38b: FT – IR spectrum for fraction of A. lanata (AL IL F1 – A. lanata in vitro leaf fraction 1)
Figure 39a: FT – IR spectrum for crude sample of A. lanata (AL MT CA – A. lanata methanol callus)
Figure 39b: FT – IR spectrum for fraction of A. lanata (AL CA F1 – A. lanata callus fraction 1)
Re
lative A
bu
nd
an
ce
100 313.23
90
80
70
60
50
40 212.08
30
20
251.28
10 198.08 359.19
180.17
0
298.26
387.36
456.72 512.02 56 0.17 637.60 670.74 751.67
824.77
885.67 960.52
100 200 300 400 500 600 700 800 900 1000
m/z
Figure 42a: LC - MS spectrum of fractions of A. javanica (AJ-FL-F1 – A. javanica field leaf fraction 1)
Re
lative A
bu
nd
an
ce
100 251.26
90
80
70
60
50
40
30
208.23 20
180.26 10
313.22
149.09 276.48 334.99 0
404.58 462.69
560.96
652.34 707.92
752.18 827.88
917.04
981.35
100 200 300 400 500 600 700 800 900 1000
m/z
Figure 42b: LC - MS spectrum of fractions of A. javanica (AJ-IL-F1 – A. javanica in vitro leaf fraction 1)
Re
lative A
bu
nd
an
ce
100 299.23
90
80
70
60
50
40
30
269.23
20
10
139.09 0
241.22
221.04
371.60
447.48
489.28
615.37
664.76
756.99
837.93
910.26 961.53
100 200 300 400 500 600 700 800 900 1000
m/z
Figure 42c: LC - MS spectrum of fractions of A. javanica (AJ-CA-F1 – A. javanica callus fraction 1)
Re
lative A
bu
nd
an
ce
100 251.25
90
80
70
60
50
40
30
20
10
149.11
0
208.24
236.19 180.28
283.21 343.20
391.09
447.34 469.15
523.17
566.56 5 88.00 6 26.48
100 150 200 250 300 350 400 450 500 550 600
m/z
Figure 43a: LC - MS spectrum of fractions of A. lanata (AL-FL-F1 – A. lanata field leaf fraction 1)
Re
lative A
bu
nd
an
ce
100 343.22
90
80
70
60
50
40
30
20
10
192.13 0
313.22
285.21
256.31
234.84
391.12 430.95
513.69 579.33
649.31
697.69
759.82
831.30 916.06
985.99
100 200 300 400 500 600 700 800 900 1000
m/z
Figure 43b: LC - MS spectrum of fractions of A. lanata (AL-IL-F1 – A. lanata in vitro leaf fraction 1)
Re
lative A
bu
nd
an
ce
100 439.18
90
80
70
60
50
40
30
20
212.09 10
198.09 274.36 313.26 424.23
489.14 712.96
755.05
560.94 0
622.40 788.89 907.73 999.17
100 200 300 400 500 600 700 800 900 1000
m/z
Figure 43c: LC - MS spectrum of fractions of A. lanata (AL-CA-F1 – A. lanata callus fraction 1)
REFERENCES
Abbas JA, El-Oqlah AA, Mahasne AM (1992). Herbal plants in the traditional
medicine of Bahrain. Econ Bot 46: 158 – 163.
Abdin MZ, Ilah A (2007). Plant regeneration and somatic embryogenesis from stem
and petiole explants of Indian chicory (Cichorium intybus L.). Ind J Biotech
6: 250 – 255.
Abhishek B, Rohini S, Satnam S, Malleshappa NN (2012). Antioxidant Activity of
Various Fractions of Ethanol Extract on Aerva lanata Linn. Planta Activa 1:
86 – 89.
Adam M, Tasarz P, Szypula E (2008). Antioxidant activity of herb extracts from
five medicinal plants from Lamiaceae, subfamily Lamioideae. J Medicinal
Plants Res 2: 321 – 330.
Agostino CB, MJ Salvador; DA Dias, MD Baruffi, LS Pereira Crott (2008)
Evaluation of immunomodulatory and anti-inflammatory effects and
phytochemical screening of Alternanthera tenella Colla (Amaranthaceae)
aqueous extracts. Mem INST Oswaldo Cruz, Rio De Janeiro, 103 569 – 577.
Ahmad N, Fazal H, Zamir R, Khalil SA, Abbasi BH (2011). Callogenesis and shoot
organogenesis from flowers of Stevia rebaudiana (Bert.) Sugar Tech 13: 174
– 177.
Ahmed E, Imran M, Malik A, Ashraf M (2006). Antioxidant activity with
flavonoidal constituents from Aerva persica. Arch Pharm Res 29: 343 – 347.
Ahmed EHM, Bakri YMN, Yousif G. Mohammed, Hassan SK (2010).
Antiplasmodial Activity of Some Medicinal Plants Used in Sudanese Folk-
medicine. Environ Health Insights 4: 1 – 6.
Aiyar V, Narayanan V, Seshadri TR, Vydeeswaran S (1973). Chemical components
of some Indian medicinal plants. Ind J Chem 11: 89 – 90.
Ajila CM, Jaganmohan Rao L, Prasada Rao UJS (2010). Characterization of
bioactive compounds from raw and ripe Mangifera indica L. peel extracts.
Food Chem Toxicol 48: 3406 – 3411.
Alagesan K, Prabhakar T, Vinoth K, Sadasivam S, Thayumanavan B (2012).
Identification of α-Glucosidase inhibitors from Psidium guajava Leaves and
Syzygium cumini Linn. Seeds. Int J Pharma Sci Res 3: 316 – 322.
i
ii
Ali Mansoori G, Kenneth S, Brandenburg, Ali Shakeri Z (2010). A Comparative
Study of Two Folate-Conjugated Gold Nanoparticles for Cancer
Nanotechnology Application. Cancers 2: 1911 – 1928.
Ali P, Elmira S, Katayoun J (2010). Comparative study of the antibacterial,
antifungal and antioxidant activity and total content of phenolic compounds
of cell cultures and wild plants of three endemic species of Ephedra.
Molecules 15: 1668 – 1678.
Almeida C, Azevedo NF, Bento F et al., (2013). Rapid detection of urinary tract
infections caused by Proteus spp. using PNA-FISH. Eur J Clinical Microbiol
Infectious Dis 32: 781 – 786.
Alveera S, Kandasamy T, Bharti O (2009). In vitro propagation of Alternanthera
sessilis (sessile joyweed), a famine food plant. Afr J Biotech 8: 5691 – 5695.
Amoo SO, Aremu AO, Van Staden J (2012). In vitro plant regeneration, secondary
metabolite production and antioxidant activity of micropropagated Aloe
arborescens Mill. Plant Cell Tiss Org 111: 345 – 358.
Anand SP, Velmurugan G, Keerthiga M, Nandagopalan V (2014). Comparative
study on Antimicrobial Activity of Argemone mexicana Linn. and Aerva
javanica (Burm.F.) Juss. Ex. Schult. Am J Pharm Tech Res 4: 530 – 540.
Ang Lee MK, Moss EJ, Yuan CSP (2001) Herbal medicines and preoperative care.
J Am Medical Association 286: 208 – 216.
Anita A, Malar Retna A (2013). Review on the medicinal plant - Aerva Lanata.
Asian J Biochem Pharma Res 3: 215 – 224.
Antoaneta I, Ivajla D, Iva T, Atanas K, Ivanka K (2002). GC-MS analysis and anti-
microbial activity of acidic fractions obtained from Paeonia peregrine and
Paeonia tenuifolia roots. Z. Naturforsch 57: 624-628.
Aravindaram K, Yang NS (2010) Anti-inflammatory plant natural products for
cancer therapy. Planta Medica 76: 1103 – 1117.
Ariffin SHZ, Wan Omar WH, Ariffin ZZ, Safian MF, Senafi S, Abdul Wahab RM
(2009). Intrinsic anticarcinogenic effects of Piper sarmentosum ethanolic
extract on a human hepatoma cell line. Cancer Cell Int 9: 6.
3
Arikat NA, Jawad FM, Karam NS, Shibli RA (2004). Micropropagation and
accumulation of essential oils in wild sage (Salvia fruticosa Mill.). Scientia
Hort 100: 193 – 202.
Arockiasamy DI, Muthukumar B, Natarajan E, John Britto S (2002). Plant
regeneration from node and internode explants of Solanum trilobatum L.
Plant Tissue Cult 12: 93 – 97.
Arumugam M, Ramachandra PU, Gomathinayagam M, Murugan G, Lakshmanan A,
Panneerselvam R (2012). Antibacterial and antioxidant activity between
micropropagated and field grown plants of Excoecaria agallocha L. Int Res J
Pharm 3: 235 – 241.
Arun M, Subramanyam K, Theboral J, Ganapathi A, Manickavasagam M (2014).
Optimized shoot regeneration for Indian soybean: the influence of exogenous
polyamines. Plant Cell Tiss Org 117: 305 – 309.
Ashraf K, Mujeeb M, Altaf A, Mohd Amir, Nasar M, Deepak S (2012). Validated
HPTLC analysis method for quantification of variability in content of
curcumin in Curcuma longa L (turmeric) collected from different
geographical region of India. Asian Pacific J Trop Biomed 584-588.
Awaad SA, Al-Jaber AN (2010). Antioxidant natural plant. In: Govil JN, Singh VK.
(Eds.) In: RPMP Ethnomedicine: Source and Mechanism. Studium Press,
India. 7: 1–35.
Ayyanar M, Sankarasivaram K, Ignacimuthu S, Sekar T (2008). Plant species with
ethanobotanical importance other than medicinal in Theni District of Tamil
Nadu, Southern India. Asian J Exp Biol 1: 765 – 771.
Baati L, Fabre-Gea C, Auriol D, Blanc PJ (2000). Study of the cryotolerance of
Lactobacillus acidophilus: effect of culture and freezing conditions on the
viability and cellular protein levels. Int J Food Microbiol 59:241-247.
Bairu MW, Amoo SO, Van Staden J (2010). Comparative phytochemical analysis of
wild and in vitro derived greenhouse grown tubers, in vitro shoots and
callus-like basal tissues of Harpagophytum procumbens. South Afr J Botany
77: 479 – 484.
Balcht, Aldona, Smith, Raymond (1994). Pseudomonas aeruginosa: Infections and
Treatment. Informa Health Care 83 – 84.
4
Basha SD, Sujatha M (2007). Inter and intra population variability of Jatropha
curcus L. characterizaed by RAPD and ISSR markers and development of
population – specific SCAR markers. Euphytica 156: 375 – 386.
Baskaran P, Jayabalan N (2005). An Efficient Micro propagation of system for
Eclipta alba – a valuable medicinal herb, In vitro Cell Dev Biol Plant 41:
532 – 539.
Battu GR, Kumar BM (2012). In vitro Antioxidant Activity of Leaf Extract of Aerva
lanata Linn. Int J Pharm Sci 4: 74 – 78.
Battu PR, Reddy MS (2009). Isolation of secondary metabolites from Pseudomonas
fluorescens and its characterization. Asian J Res Chem 2: 26 – 29.
Baum BR, Mechanda SM, Livesey JF, Binns SE, Arnason JT (2001). Predicting
quantitative phytochemical markers in single Echinacea plants or clones
from their DNA fingerprints. Phytochem 56: 543 – 549.
Bedi SJ, Patel (1978). Ethanobotany of the Ratan Mahal Hills, Gujarat, India. Econ
Bot 15: 145 – 152.
Bekada AMA, Benakriche B, Hamadi K, Bensoltane A (2008). Modelling of effect
of water activity, pH and temperature on the growth rate of Mucor
racemousus isolated from soft camembert cheese. World J Agri Sci 4: 790 –
794.
Besser RE, Lett SM, Weber JT, Doyle MP, Barrett TJ, Wells JG, Griffin PM (1993).
An outbreak of diarrhoea and haemolytic uremic syndrome from Escherichia
coli O157:H7 in fresh-pressed apple cider. J Am Medical Association 269:
2217 – 2220
Bhanot A, Sharma R, Singh S, Noolvi MN, Singh S (2013). In vitro anticancer
activity of ethanol extract fractions of Aerva lanata L. Pak J Biol Sci 16:
1612 – 1617.
Bhatia A, Manju A, Rupali S, Sharma A (2013). Antioxidant activity of native and
micropropagated Tylophora indica leaves extract: A comparative study.
J Nat Prod Plant Resour 3: 1 – 7.
Bhatia R, Singh KP, Sharma TR, Jhang T (2011). Evaluation of the genetic fidelity
of in vitro propagated gerbera (Gerbera jamesonii Bolus) using DNA-based
markers. Plant Cell Tiss Org 104: 131-135.
5
Bhojwani SS, Dantu PK (2013). Plant Tissue Culture: An Introductory Text.
Springer, New York.
Bibi Y, Zia M, Nisa S, Habib H, Waheed A, Chaudhary FM (2011). Regeneration of
Centella asiatica plants from non- embryogenic cell lines and evaluation of
antibacterial and antifungal properties of regenerated calli and plants. J Biol
Eng 5:13 – 18.
Blios MS (1958). Antioxidant’s determination by the use of a stable free radical.
Nature 4617: 1199-1200.
Boros B, Farkas A, Jakabova S, Bacskay I, Kilar F, Felinger A (2010). LC-MS
Quantitative Determination of Atropine and Scopolamine in the Floral
Nectar of Datura Species. Chromatographia 71: 43-49.
Botstein B, White RL, Skolnick M, Davis RW, Hum AJ (1980). Construction of a
genetic linkage map in man using restriction fragment length
polymorphisms. Genetics 32: 314 – 331.
Bougard F, Gravot A, Milesi S, Gontier E (2001). Production of plant secondary
metabolites: A Historical perspective. Plant Sci 161: 839 – 851.
Bravo L (1998). Polyphenols: Chemistry, dietary sources, metabolism, and
nutritional significance. Nutr Rev 56: 317-333.
Brett D, Shepard, Gilmore MS (2002). Differential expression of virulence related
genes in Enterococcus faecalis in response to biological cues in serum and
urine. Infect Immun 70: 4344 – 4352.
Brusotti G, Cesari I, Dentamaro A, Caccialanza G, Massolini G (2014). Isolation
and characterization of bioactive compounds from plant resources: The role
of analysis in the ethnopharmacological approach. J Pharma Biomed
Analysis 87: 218 – 228.
Bui van Le, Do my NT, Jeanneau M, Sadik S, Shanjun T et al., (1998). Rapid plant
regeneration of a C4 dicot species: Amaranthus edulis. Plant Sci 132: 45 – 54.
Cai Y, Luo Q, Sun M, Corke H (2004). Antioxidant activity and phenolic
compounds of 112 traditional Chinese medicinal plants associated with
anticancer. Life Sci 74: 2157 – 2184.
6
Cai YZ, Sun M, Xing J, Luo Q, Corke H (2006). Structure radical scavenging
activity relationships of phenolic compounds from traditional Chinese
medicinal plants. Life Sci 78: 2872 – 2888.
Cai Z, Lee FSC, Wang XR, Yu WJ (2002). A capsule review of recent studies on the
application of mass spectrometry in the analysis of Chinese medicinal herbs.
J Mass Spect 37: 1013 – 1024.
Cannell RJP (1998). Natural Products Isolation. New Jersey: Human Press Inc.
165 – 208.
Carra A , Maurizio S, Loredana A, Mirko S, Francesco S, Francesco C (2012). In vitro
plant regeneration of caper (Capparisspinosa L.) from floral explants and
genetic stability of regenerants. Plant Cell Tiss Org 109: 373 – 381.
Castello M, Phatak A, Chandra N, Sharon M (2002). Antimicrobial activity of crude
extracts from plant parts and corresponding calli of Bixa orellana L. Indian J
Exp Biol 40: 1378 – 1381.
Ceasar SA, Maxwell SL, Prasad KB, Karthigan M, Ignacimuthu S (2010). Highly
efficient shoot regeneration of Bacopa monnieri L. using a two-stage culture
procedure and assessment of genetic integrity of micropropagated plants by
RAPD. Acta Physiol Plant 32: 443 – 452.
Center for Disease control and Prevention (CDC) Staphylococcus aureus Resistant
to Vancomycin - United States, 2002. Morbidity Mortality Weekly Rep 51:
565 – 567.
Chaleshtori SB, Behrouz S, Masoomeh K et al., (2012). Assessment of genetic
diversity and structure of Imperial Crown (Fritillaria imperialis L.)
populations in the Zagros region of Iran using AFLP, ISSR and RAPD
markers and implications for its conservation. Biochemical Systematics and
Ecology 42: 35 – 48.
Chan K (2003). Some aspects of toxic contaminants in herbal medicines.
Chemosphere 52: 1361 – 1371.
Chandra S, Sastry MS (1990). Chemical constituents of Aerva lanata. Fitoterapia
61: 188.
vii
Chapman MR, Robinson LS, Pinkner JS, Roth R, Heuser J, Hammar M, Normark S,
Hultgren SJ (2002). Role of Escherichia coli curli operons in directing
amyloid fiber formation. Science 295: 851 – 855.
Chelli CN, Tir Touil Meddah A, Mullie C, Aoues A, Meddah B (2012). In vitro and
in vivo antimicrobial activity of Algerian Hoggar Salvadora persica L.
extracts against microbial strains from children's oral cavity.
J Ethnopharmacol 144: 57 – 66.
Chetrisiva PK, Sharma K, Veena A (2013). Genetic diversity analysis and screening
of high psolralen yielding chemotype of Psoralea corylifolia from
different regions of India employing HPLC and RAPD marker. Int J Plant
Res 26: 88 – 95.
Choma IM, Grzelak EM (2011). Bio autography detection in thin-layer
chromatography. J Chromatography 1218: 2684 – 2691.
Chopra RN, Nayar SL, Chopra IC (1956). Glossary of Indian Medicinal Plants.
CSIR, New Delhi: 194.
Christen Y, Olano-Martin E, Packer L (2002). EGb-761 in the postgenomic era: new
tools from molecular biology for the study of complex products such as
Ginkgo biloba extract. Cell Mol Biol 48: 593 – 539.
Claydon N, Grove JF Pople M (1985). Elm bark beetle boring and feeding
detergents from Phomopsis oblanga. Phytochem 24: 937 – 943.
Coldren CD, Hashim P, Ali JM, Oh SK, Sinskey AJ, Rha C (2003). Gene expression
changes in the human fibroblast induced by Centella asiatica triterpenoids.
Planta Medica 69: 725 – 732.
Cole RB (1997). Electron spray Ionization Mass Spectrometry: Fundamentals,
Instrumentation and Applications. Hoboken, NJ USA.
Cook NC, Samman S (1996). Flavonoids- chemistry, metabolism, cardioprotective
effects, and dietary sources. Nutritional Biochem 7: 66 – 76.
Costa P, Goncalves S, Valentao P, Andrade PB, Coelho N, Romano A (2012).
Thymus lotocephalus wild plants and in vitro cultures produce different
profiles of phenolic compounds with antioxidant activity, Food Chemistry
135: 1253 – 1260.
8
Costa PS, Valentao GP, Paula B, Coelho AN, Roman A (2012) Thymus lotocephalus
wild plants and in vitro cultures produce different profiles of
phenolic compounds with antioxidant activity. Food Chem
135: 1253 – 1260.
Cragg GM, Newman DJ (2005) Plants as a source of anti-cancer agents.
J Ethnopharmacol 100: 72 – 79.
Dai J, Mumper RJ (2010). Plant phenolics: Extraction, analysis and their antioxidant
and anticancer properties. Molecules 15: 7313 – 7352.
Dakah AK, Salim Z, Mohamad S, Sami A, Wink M (2014). In vitro propagation of
the medicinal plant Ziziphora tenuior L. and evaluation of its antioxidant
activity. Saudi J Biol Sci 21: 317 – 323.
Das SS, Gauri SS, Misra BB, Biswas M, Dey S (2013). Purification and
characterization of a betanidin glucosyl transferase from Amaranthus tricolor
L catalyzing non-specific biotransformation of flavonoids. Plant Sci 211:
61– 69.
Devarumath RM, Nandy S, Rani V, Marimuthu S, Muraleedharan N (2002). RAPD,
ISSR and RFLP fingerprints as useful markers to evaluate genetic integrity
of micropropagated plants of three diploid and triploid elite tea clones
representing Camellia sinensis (China type) and C. assamica ssp. assamica
(Assam- India type). Plant Cell Rep 21:166 – 173.
Devi JR, Thangam EB (2010). Extraction and separation of glucosinolates from
Brassica oleraceae var rubra. Advanced Biomed Res 4: 309 – 313.
Dias ACP, Seabra RM, Andrade PB, Fernandes FM (1999). The development and
evaluation of an HPLC-DAD method for the analysis of the phenolic
fractions from in vivo and in vitro biomass of Hypericum species. J Liquid
Chromatogr 22: 215 – 227.
Dinis TCP, Madeira VMC, Almeida LM (2004). Action of phenolic derivatives
(acetoaminophen, salycilate and 5-aminosalycilate) as inhibitors of
membrane lipid peroxidation and as peroxyl radical scavengers. Arch
Biochem Biophy 315: 161 – 169.
9
Dolendro S, Lingaraj S, Neera BS, Pawan KJ (2003). The effect of TDZ on
organogenesis and somatic embryogenesis in pigeon pea (Cajanus cajan L.
Mill). Plant Sci 164: 341 – 347.
Donnell GO, Bucar F, Gibbons S (2006). Phytochemistry and antimycobacterial
activity of Chlorophytum incornatum. Phytochem 67: 178 – 182.
Dulay C, Sayeeda AI Alam Bhuiyan MS, Astaq MK (2002). Antimicrobial activity
and cytotoxicity of Aerva lanata. Fitoterapia 73: 92 – 94.
Duraipandiyan V, Ayyanar M, Ignacimuthu S (2006). Antimicrobial activity of
some ethnomedicinal plants used by Palyar tribe from Tamil Nadu, India.
BMC Complement. Altern Med 6: 35 – 41.
Early Breast Cancer Trialists Collaborative Group (EBCTCG) (2005). Effects of
chemotherapy and hormonal therapy for early breast cancer on recurrence
and 15-year survival: An overview of the randomised trials. Lancet 365:
1687 – 1717.
Eberhardt TL, Li X, Shupe TF, Hse CY (2007). Chinese Tallow Tree (Sapium
sebiferum) utilization: Characterization of extractives and cell wall
chemistry. Wood Fiber Sci 39: 319 – 324.
Eemay L, Laikeng C, Boey PL (2003). Tropane alkaloids in micropropagated plants
and callus of Hyoscyamus niger. J Trop Med Plants 4: 103 – 108.
Egwaikhide PA and Gimba CE (2007). Analysis of the Phytochemical Content and
Anti-microbial Activity of Plectranthus glandulosis Whole Plant. Middle-
East J Sci Res 2: 135 – 138.
Egwaikhide PA, Okeniyi SO, Gimba CE (2007). Screening for antimicrobial activity
and phytochemical constituents of some Nigerian medicinal plants. Adv Biol
Res 1: 155 – 158.
Ehsan O, Norhani A, Syahida A, Wan ZS, Abdul RO, Yin WH (2011). Bioactive
Compounds and Biological Activities of Jatropha curcas L. Kernel Meal
Extract. Int J Mol Sci 12: 5955 – 5970.
El-Hadi MA, Barki YMN, Yousif GM, Hassan SK (2010). Antiplasmodial activity
of some medicinal plants used in Sudanese folk-medicine. Environ Health
Insight 4: 1 – 6.
10
Eloff JN (1998). A sensitive and quick microplate method to determine the
minimum inhibitory concentration of plant extract for bacteria. Planta Med
64: 711 – 714.
Emam SS (1999). Phytochemical studies on the herb Aerva javanica growing in
Egypt. Cairo University Faculty of Agriculture Bulletin 50: 488 – 514. Ennajar M,
Bouajila J, Lebrihi A, Mathieu F, Abderraba M, Raies A, Romdhane M
(2009). Chemical composition and antimicrobial and antioxidant activities of
essential oils and various extracts of Juniperus phoenicea L. (Cupressacees)
J Food Sci 74: 364 – 371.
Entaz B, Joushan A, Mofasser H, Raihan O (2013). Antioxidant (In vitro) Effect of
Methanol and Petroleum Ether Extracts of the Aerva lanata. Pharma
Innovation J 2: 36 – 43.
Espin JC, Conesa GMT, Barberan FA (2007). Nutraceuticals: facts and fiction.
Phytochem 68: 2986 – 3008.
Farees UDM, Hanif U, Asia B et al., (2012). Antimicrobial activities of Aerva
javanica and Paeonia emodi plants. Pak. J Pharm Sci 25: 565 – 569.
Fatimi ALM, Wurster M, Schroder G, Lindequist U (2007). Antioxidant,
antimicrobial and cytotoxic activities of selected medicinal plants from
Yemen. J Ethnopharmacol. 111: 657 – 666.
Fatinah AA, Arumingtyas EL, Mastuti R (2012). Genetic diversity study among six
genera of Amaranth family found in Malang based on RAPD Marker.
J Tropical Life Sci 2: 81 – 86.
Fico G, Spada A, Braca A, Agradi E, Morelli I, Tome F (2003). RAPD analysis and
flavonoid composition of Aconitum as an aid for taxonomic discrimination.
Biochem Syst Ecol 31: 293 – 301.
Fink SL, Cookson BT (2005) Apoptosis, pyroptosis, and necrosis: mechanistic
description of dead and dying eukaryotic cells. Infection Immu 73: 1907 –
1916.
Fulda S (2010) Modulation of apoptosis by natural products for cancer therapy.
Planta Medica 76: 1075 – 1079.
11
Gantet P, Memelink J (2002). Transcription factors: tools to engineer the
production of pharmacologically active plant metabolites. Trends Pharma
Sci 23: 563 – 569.
Garg SP, Bhushan R, Mehta R, Jain VM, Dutta BK, Indrani J (1980). A survey for
alkaloids in Rajasthan desert plants. Trans Indian Soc Desert Tecnol Univ
Cent Desert Stud 5: 62 – 64.
Gebauer M (2004). Microarray applications: Emerging technologies and
perspectives. Drug Discovery Today 9: 915 – 917.
Ghazanfar SA (1994). Hand book of Arabian Medicinal Plants. – M. Sc. Thesis. Bot
Dept Fac Sci Al-Azhar Univ.
Gil MI, Barberan FAT, Pierce BH, Holcroft DM, Kader AA (2000). Antioxidant
Activity of Pomegranate Juice and Its Relationship with Phenolic
Composition and Processing. J Agric Food Chem 48: 4581 – 4589.
Gilbert RJ, Turnbull PCB, Parry JM, Kramer JM (1981). Bacillus cereus and other
Bacillus species: Their part in food poisoning and other clinical infections,
In: R.C.W. Berkeley and M. Goodfellow (ed.), The Aerobic Endospore
forming Bacteria. Academic Press Inc., London. 297 – 314.
Girach RD, Aminuddin SPA, Khan SA, (1994). Traditional plant remedies among
the Kondh of district Dhenkal Orissa. Int J Pharma 32: 274 – 283.
Girija K, Kuruba L, Nagaraj P, Pulla UC (2011). Antihyperglycemic and
hypolipidemic activity of methanolic extract of Amaranthus viridis leaves in
experimental diabetes. Indian J Pharmacol 43: 450 – 454.
Girish HV, Sudarshana MS, Rao ER (2008). In vitro evaluation of the efficacy of
leaf and its callus extracts of Cardiospermum halicacabum L. on important
human pathogenic bacteria. Adv Biol Res 2: 34-38.
Goel MK, Kukreja AK, Bisht NS (2009). In vitro manipulations in St. John’s wort
(Hypericum perforatum L.) for incessant and scale up micropropagation
using adventitious roots in liquid medium and assessment of clonal fidelity
using RAPD analysis. Plant Cell Tiss Org 96: 1 – 9.
Gopalakrishnan S Vadivel E (2011). GC-MS analysis of some bioactive constituents
of Mussaenda frondosa Linn. Int J Pharma Bio Sci 2: 313 – 320.
xii
Goto CE, Barbosa EP, Kistner LC, Moreira FG, Lenartovicz V, Peralta RM (1998).
Production of amylase from Aspergillus fumigatus utilizing alpha-methyl-D-
glycoside, a synthetic analogue of maltose, as substrate. Microbiol Lett 167:
139 – 143.
Goyal M, Anil P, Nagori BP, Sasmal D (2011). Aerva lanata: A review on
phytochemistry and pharmacological aspects. Phcog Rev 5: 195 – 198.
Griffith DWT, Deutscher NM, Caldow C, Kettlewell G, Riggenbach M, Hammer S
(2012). A Fourier transform infrared trace gas and isotope analyser for
atmospheric applications. Atmos Meas Tech 5: 2481 – 2498.
Gross JG (2004). Mass Spectrometry. Springer-Verlag, New York, NY USA.
Gujjeti RP, Estari M (2013). Anti-HIV activity and cytotoxic effects of Aerva lanata
root extracts. Am J Phytomed Clinical Therapeutic 2: 894 – 900.
Gulcin I, Oktay MO, et al., (2002). Determination of antioxidant activity of lichen
Cetraria islandica (L) Ach. J Ethnopharmacol 79: 325 – 329.
Gulshan C, Darshna C, Madan V, Manish S, Pawan KJ (2008). TDZ-induced direct
shoot organogenesis and somatic embryogenesis on cotyledonary node
explants of lentil (Lens culinaris Medik.). Physiology Molecular Biol Plants
14: 347-353.
Guvenalp Z, Demirezer LO (2005). Flavonol glycosides from Asperula arvensis.
Turkish J Chem 29: 163 – 169.
Hale TL, Formal SB (1981). Protein synthesis in HeLa or Henle 407 cells infected
with Shigella dysenteriae 1, Shigella flexneri 2a, or Salmonella typhimurium
W118. Infect. Immun 32: 137-144.
Hanumantharaju N, Shashidhara S, Rajasekharan PE, Rajendra CE (2010).
Comparative evaluation of antimicrobial and antioxidant activities of
Kaempferia galanga for natural and micropropagated plant. Int J Pharm
Pharm Sci 2: 72 – 75.
Harborne JB (1984). A Guide to Modern Techniques of Plant Analysis. Chapman
and Hall, London. 4 – 80.
Harborne JB (1989). General procedures and measurement of total phenolics.
Methods in plant biochemistry: Volume 1 Plant Phenolics, Academic Press,
London. 1 – 28.
13
Harborne JB (1999). Phytochemical dictionary: Handbook of bioactive compounds
from plants 2nd (Edn.). Taylor and Francis, London. 221-234.
Harrison AG (1992). Chemical Ionization Mass Spectrometry. 2nd ed., CRC Press,
Boca Raton, FL USA.
Hashimoto K, Higuchi M, Makino B, Sakakibara I, Kubo M, Komatsu Y, Maruno M,
Okada M (1999). Quantitative analysis of aristolochic acids, toxic compounds,
contained in some medicinal plants. J Ethanopharmacol 64: 185 – 189.
Hassanein RA, Hashem HA, Khalil RR (2012) Stigmasterol treatment increases salt
stress tolerance of faba bean plants by enhancing antioxidant systems. Plant
Omics J 5:476-485.
Hazra K, Roy RN, Sen SK, Laska S (2007). Isolation of antibacterial pentahydroxy
flavones from the seeds of Mimusops elengi Linn. Afr J Biotech. 6: 1446 –
1449.
He XG (2000). On-line identification of phytochemical constituents in botanical
extracts by combined high-performance liquid chromatographic-diode array
detection mass spectrometric techniques. J Chromatography A 880: 203 – 232.
Hedge JE, Hofreiter BT (1962). In: Carbohydrate Chemistry, 17 (Eds. Whistler R.L.
and Be Miller JN), Academic Press, New York.
Hsin Ling Y, Senthil Kumar KJ, You-Cheng H (2012). Multiple molecular targets of
Antrodia camphorata: A suitable candidate for breast cancer
chemoprevention, targeting new pathways and cell death in breast cancer,
Dr. Rebecca Aft (Ed.) 98.
Hvattum E (2002). Determination of phenolic compounds in rose hip (Rosa canina)
using liquid chromatography coupled to electrospray ionisation tandem
mass spectrometry and diode-array detection. Rapid Communic Mass Spec
16: 655 – 662.
Iizuka N, Oka M, Yamamoto K, Tangoku A, Miyamoto K et al., (2003).
Identification of common or distinct genes related to antitumor activities of a
medicinal herb and its major component by oligonucleotide microarray. Int J
Cancer 107: 666 – 672.
Ingale AG, Hivrale AU (2010). Pharmacological studies of Passiflora sp and their
bioactive compounds. Afr J Plant Sci 4: 417 – 426.
14
Ito Y (2005). Golden rules and pitfalls in selecting optimum conditions for high-
speed counter current chromatography. J Chromatogr A 1065: 145 – 168.
Jacob A, Malpathak N (2004). Green hairy roots of Solanum khasianum Clarke: A
new route to in vitro solasodine production. Current Science 87: 1442 –
1447.
Jain SM (2001). Tissue culture derived variation in crop improvement. Euphytica
118: 153 – 166.
Jayasri MA, Mathew L, Radha A (2009). A report on the antioxidant activities of
leaves and rhizomes of Costus pictus D. Don. Int J Integr Biol 5: 20 – 26.
Jemal A, Bray F, Center MM, Ferlay J (2011). Global Cancer Statistics. CA: Cancer
J Clin 61: 69 – 90.
Joanofarc J, Vamsadhara C (2003). Evaluation of antidiarrhoeal activity of Aerva
species. Nat Prod Sci 9: 177 – 179.
Johnson M, Petchiammal E, Janakiraman N, Babu A, Renisheya JJ, Malar T,
Sivaraman A (2012). Phytochemical characterization of brown seaweed
Sargassum wightii. Asian Pacific J Trop Dis 2: 109 – 113.
Johnson M, Wesely EG, Zahir Hussain MI, Selvan N (2010). In vivo and in vitro
phytochemical and antibacterial efficacy of Baliospermum montanum
(Willd.) Muell. Arg. Asian Pacific J Tropical Med 3: 894 – 897.
Jothy SL, Zakaria Z, Chen Y, Lau YL, Latha LY, Shin LN, et al., (2011). Bioassay
directed isolation of active compounds with antiyeast active from a Cassia
fistula seed extracts. Molecules 16: 7583 – 7592.
Judd WS, Campbell CS, Kellogg E, Stevens A et al., (2008). Plant Systematics: A
Phylogenetic Approach. Third Edition. Sinauer Associates, Inc. Sunderland,
MA 168.
Justesen U, Knuthsen P (2001). Composition of flavonoids in fresh herbs and
calculation of flavonoid intake by use of herbs in traditional Danish dishes.
Food Chem 73: 245 – 250.
Justesen U, Knuthsen P, Leth T (1998). Quantitative analysis of flavonols, flavones
and flavanones in fruits, vegetables and beverages by high performance
liquid chromatography with photo-diode array and mass spectrometric
detection. J Chromatography 799: 101–110.
15
Kalirajan A, Narayanan KR, Ranjitsingh AJA, Ramalakshmi C, Parvathiraj P
(2013). Bioprospecting medicinal plant Aerva lana Juss ex. Schult flowers
for potential antimicrobial activity against clinical and fish borne pathogens.
Ind J Nat Prod Res 4: 306 – 311.
Kalpana J, Chavan P, Warude D, Patwardhan B (2004). Molecular markers in herbal
drug technology. Curr Sci 87: 159 – 166.
Kamalanathan D, Natarajan D (2013). Micro propagation of Shoot tip Explants of
Aerva lanata (L). A.L. Juss. Ex.Schultes. a Resourceful Medicinal Plant,
Scientific Basis of Herbal Medicine, Daya Publishers, New Delhi 141 – 148.
Kamalanathan D, Ragavendran C, Natarajan D (2014). Antioxidant and HPLC
profile of wild and micropropagated Aerva lanata (Linn.) Juss. Ex. Schult. -
A Comparative Study. Indo Am J Pharm Res 4: 2628 – 2636.
Keivani M, Ramezanpour SS, Soltanloo H, Choukan R, Naghavi M, Ranjbar M
(2010). Genetic diversity assessment of alfalfa (Medicago sativa L.)
populations using AFLP markers. Aust J Crop Sci 4: 491 – 497.
Kenneth RM, Lawrence JP (1975). Evidence of biosynthetic simplicity in the
flavonoid chemistry of the ricciaceae. Phytochem 14: 199 – 201.
Kevers C, Le Gal N, Monteiro M, Dammers J, Gasper T (2000). Somatic
embryogenesis of Panax ginseng in liquid cultures: A role for amino acids
and their metabolic pathways. J Plant Growth Regul 31: 209 – 214.
Khachik F, Beecher GR, Whittaker NF (1986). Separation, identification, and
quantification of the major carotenoid and chlorophyll constituents in
extracts of several green vegetables by liquid chromatography. J Agri Food
Chem 34: 603 – 616.
Khalil SA, Zamir R, Ahmad N (2014). Selection of suitable propagation method for
consistent plantlets production in Stevia rebaudiana (Bertoni). Saudi J Biol
Sci, http://dx.doi.org/10.1016/j.sjbs.2014.02.005.
Khalil SA, Zamir R, Ahmad N, Sajid M, Fazal H, Khan MA, Seema N, Alam R
(2011). In vitro regeneration of plantlets from unpollinated ovary
culture in sweet orange (Citrus sinensis L. Osbeck). Afr J Biotech 10:
15130 – 15134.
16
Khan AW, Saleem Jan, Shaista P, Rahmat AK, Asma Saeed, Abdul JT, Anwar Ali
Shad (2012). Phytochemical analysis and Enzyme Inhibition Assay of Aerva
javanica for Ulcer. Chemistry Central J 6: 76 – 82.
Khan UG, Nazir T, Ahmed VU (1982). Chemical Constituents of Aerva javanica.
Fitoterapia 53: 75 – 77.
Khare CP (2007). Indian medicinal plants: An Illustrated Dictionary. Springer, New
York, 22 – 24.
Kidgella C, Reichard U, Waina J et al., (2002). Salmonella typhi, the causative agent
of typhoid fever is approximately 50,000 years old. Infection Genetics Evol
2: 39 – 45.
Kishor R, Devi HS (2009). Induction of multiple shoots in a monopodial orchid
hybrid (Aerides vandarum Reichb.f × Vanda stangeana Reichb.f) using
thidiazuron and analysis of their genetic stability. Plant Cell Tiss Org 97:
121 – 129.
Koch HM, Gonzalez-Reche LM, Angerer J (2003). On-line cleanup by
multidimensional liquid chromatography-electrospray ionization tandem
mass spectrometry for high throughput quantification of primary and
secondary phthalate metabolites in human urine. J Chromatogr B Analyt
Technol Biomed Life Sci 784:169 – 182.
Kolb CA, Kaser MA, Kopecky J, Zotz G, Riederer M, Pfundel EE (2001). Effects of
Natural Intensities of Visible and Ultraviolet Radiation on Epidermal
Ultraviolet Screening and Photosynthesis in Grape Leaves. Plant Physiol
127: 863 – 875.
Kozyrenko MM, Artyukova EV, Lauve LS, Boltenkov EV (2002). Genetic
Variability of Callus Cultures of Some Iris Species. Biotech Russia 4:
30 – 37.
Krishnamurthi A (2003) The wealth of India. Vol. I, CSIR, New Delhi. 92.
Kritikar KR, Basu BD (1996). Indian Medicinal Plants. International book
distributors. Dehradun, India 2064 – 2065.
Kucukislamoglu M, Yayli N, Senturk HB, Genc H (2000). Flavonol glycosides from
Consolida armeniaca. Turk J Chem 2: 191 – 197.
xvii
Kumar S, Pandey S, Pandey AK (2014). In vitro antibacterial, antioxidant, and
cytotoxic Activities of Parthenium hysterophorus and characterization of
extracts by LC-MS Analysis. BioMed Research Int 2014: 1 – 10.
Kumpulainen JT, Salonen JT (1999). Natural Antioxidants and Anticarcinogens in
Nutrition, Health and Disease, Royal Society Chem UK 178 – 187.
Kuzel S, Vydra J, Triska J, Vrchotova N, Hruby M, Cigler P (2009). Elicitation of
pharmacologically active substances in an intact medicinal plant. J Agri
Food Chem 57: 7907 – 7911.
Lacroix M, Leclercq G (2004) Relevance of breast cancer cell lines as models for
breast tumours: an update. Breast Research Treatment 83: 249 – 289.
Lalee A, Pal P, Bhattacharaya B, Samanta A (2012). Evaluation of Anticancer
activity of Aerva sanguinolenta (L.) (Amaranthaceae) on Ehrlich’s Ascites
cell induced Swiss Mice. Int J Drug Dev Res 4: 203 – 209.
Larkin PJ, Scowcroft WR (1981). Somaclonal variation - a novel source of
variability from cell cultures for plant improvement. Theor Ap Genet 60: 197
– 214.
Lata H, Moraes RM, Douglas A, Scheffler BE (2002). Assessment of genetic
diversity in Podophyllum peltatum by molecular markers. In: J. Janick
and A. Whipkey (eds.), Trends in new crops and new uses. ASHS Press,
537 – 544.
Leathers TD, Sypherd PS (1985). Antimicrobial agents inducible phenotypic
multidrug resistance in the fungus Mucor racemosus. Chemother. 27:
892 – 896.
Leclerc H, Schwartzbrod L, Dei Cas E (2002). Microbial agents associated with
waterborne diseases. Crit Rev Microbiol 28: 371 – 409.
Lee KH, Xiao Z (2005). Podophyllotoxins and analogs. In: Cragg, GM, Kingston
DGI Newman DJ (Eds.) Anticancer Agents from Natural Products.
Brunner-Routledge Psychology Press, Taylor Francis Group, Boca Raton,
FL, 71 – 88.
Levieille G, Wilson G (2002). In vitro propagation and iridoid analysis of the
medicinal species Harpagophytum procumbens and H. zeyheri. Plant Cell
Rep 21: 220 – 225.
18
Li HB, Chen F (2005). Isolation and purification of baicalein, wogonin and oroxylin
A from the medicinal plant Scutellaria baicalensis by high-speed counter-
current chromatography. J Chromatograph A 1074: 107 – 110.
Li ZK, Yu SB, Lafitte HR et al., (2003). QTL environment interactions in rice. I.
Heading date and plant height. Theoret. Appl. Genet 108: 141–153.
Liao YH, Tsong MC, Wen YH (2012). Identification of two licorice species,
Glycyrrhiza uralensis and Glycyrrhiza glabra, based on separation and
identification of their bioactive components WC. Liaoa, 1. Food Chemistry
132: 2188 – 2193.
Lilia AM, Rodriguez JD, Cosio SR (2004). Analysis of essential oils from wild and
micropropagated plants of damiana (Turnera diffusa). Fitoterapia 75: 696 –
701.
Lima SGD, Jose MM, Neto, Antonia MG, Lopes Cito, Jose GM Da Costa and
Francisco A. M. Reis (2009) Monoterpenes, Sesquiterpenes and Fatty Acids
from Julocroton triqueter (Euphorbiaceae) From Ceara – Brazil J Chil Chem
Soc 54: 55-57.
Liu RH (2004). Potential synergy of phytochemicals in cancer prevention:
mechanism of action. J Nutr 134: 3479 – 3485.
Lugato D, Simao MJ, Garcia R, Mansur E, Pacheco G (2014). Determination of
antioxidant activity and phenolic content of extracts from in vivo plants and
in vitro materials of Passiflora alata Curtis. Plant Cell Tiss Org DOI
10.1007/s11240-014-0486-4.
Luisa CC, Luis G, Cristina O, Jose CG, Sara A (2003). RAPD assessment for
identification of clonal identity and genetic stability of in vitro propagated
chestnut hybrids. Plant Cell Tiss Org Cult 1 – 5.
Lyne RL, Mulheirn LJ, Leworthy DP (1976). New pterocarpinoid phytoalexins of
soybean. J Chem Soc Chem Commun 497 – 498.
Ma G, Jaime A, Teixeira DS, Xinhua Z, Jietang Z (2011). Shoot organogenesis and
somatic embryogenesis from leaf and shoot explants of Ochna integerrima
(Lour). Plant Cell, Tissue Organ Cult 104: 157 – 162.
19
Makkar HPS (2003). Quantification of tannins in tree and shrub foliage: A
laboratory mannual. Dondrecht, the Netherlands: Kluwer Academic
Publishers.
Malik CP (2007). Applications of Biotechnology innovations in pharmaceutics and
nutraceutics in multi therapeutic medicinal and special plants. ed Karan
Singh, ML Jahdon and D Singh. Aavishkar publishers Jaipur 2: 243 – 265.
Malik CP, Poonam G, Yaksha Singh, Staffi G (2012). Medicinal uses, chemical
constituents and micropropagation of three potential medicinal plants.
Int J life Sci Pharm Res 2: 57 – 76.
Mallabev A, Rakhimov A, Murdakhaev YM (1989). Carbohydrates of Aerva lanata.
Chem Nat Comp 25: 369 – 370.
Mander M, Mckenzie M (2005). Southern African Trade Directory of Indigenous
Natural Products. Commercial Products from the Wild Group, Matieland. 3 – 8.
Manikandan A, Victor ADA (2010). Evaluation of biochemical contents, nutritional
value, trace elements, SDS-PAGE and HPTLC profiling in the leaves of Ruellia
tuberosa L. and Dipteracanthus patulus (Jacq.). J Chem Pharm Res 2: 295 –
303.
Maruthupandian A, Mohan VR (2011) GC-MS analysis of some bioactive
constituents of Pterocarpus marsupium Roxb. Int J Chemtech Res 3: 1652.
Marzouk M, Moharram F, Mohamed M, Gamal Eldeen A, Aboutabl E (2007).
Anticancer and antioxidant tannins from Pimenta dioica leaves. Z.
Naturforsch 62: 526 – 536.
Mattson MP, Cheng A (2006). Neurohormetic phytochemicals: low dose toxins that
induce adaptive neuronal stress responses. Trends in Neurosci 29: 632 – 639.
Meirinhos J, Silva BM, Valentao P, Seabra RM, Pereira JA, Dias A, Andrade PB,
Ferreres F. Analysis and quantification of flavonoidic compounds from
Portuguese olive (Olea Europaea L.) leaf cultivars. Natural Prod Res 19:
189 – 195.
Melendez A, Rodriguez DJ, Cosio RS (2004). Analysis of essential oils from wild
and micropropagated plants of damiana (Turnera diffusa). Fitoterapia 75:
696 – 701.
20
Miliauskasa G, Venskutonis PR, Van Beek TA (2004). Screening of radical
scavenging activity of some medicinal and aromatic plant extracts. Food
Chem 85: 231 – 237.
Minocha R, Shortle WC, Long S, Minocha SC (1999). Polyamine levels during the
development of zygotic and somatic embryos of Pinus radiate. Physiol Plant
105: 155 – 164.
Modupe O, Wesley O, Edith OF, Elizabeth OA (2009). Analysis of the essential oil
from the dried leaves of Euphorbia hirta Linn (Euphorbiaceae), a potential
medication for asthma. Afr J Biotechn 8: 7042 – 7050.
Mohan N, Jassal PS, Kumar V, Singh RP (2011). Comparative in vitro and in vivo
study of antioxidants and phytochemical content in Bacopa monnieri. Recent
Res Sci Tech 3: 78 – 83.
Mohanty S, Panda MK, Subudhi E, Acharya L, Nayak S (2008). Genetic stability of
micropropagated ginger derived from axillary bud through cytophotometric
and RAPD analysis. Z Naturforsch 63:747 – 754.
Monagas M, Suarez R, Cordoves CG, Bartolome B (2005). Simultaneous
determination of nonanthocyanin phenolic compounds in red wines by
HPLC-DAD/ESI-MS. Am J Enol Vitic 56: 139 – 147.
Monga J, Pandit S, Singh Chauhan C, Sharma M (2013). Cytotoxicity and apoptosis
induction in human breast adenocarcinoma MCF-7 cells by (+)-cyanidan-3-
ol. Experimental Toxicol Pathol 65: 1091 – 1100.
Monga J, Saurabh P, Rajinder SC, Chetan SC, Shailender SC, Manu S (2013a).
Growth Inhibition and Apoptosis Induction by (+)-Cyanidan-3-ol in
Hepatocellular Carcinoma. Plos one 8: 1 – 18.
Monteiro M, Kevers C, Dommes J, Gaspar T (2002). A specific role for spermidine
in the initiation phase of somatic embryogenesis in Panax ginseng CA
Meyer. Plant Cell Tiss Org Cult 68: 225-232.
Moore CB, Sayers N, Mosquera J, Slaven J, Denning DW (2000). Antifungal drug
resistance in Aspergillus. J Infec.41: 203-220.
Moquammel SH, Biswajit G (2013). Micropropagation, in vitro flowering and
cytological studies of Bacopa chamaedryoides, an ethno-medicinal plant,
Environment Experimental Biol 11: 59 – 68.
21
Mossa JS, Al Yahya MA, Al Meshal IA (1987). Medicinal plants of Saudi Arabia,
Kingdom of Saudi Arabia. King Saud University Press. Riyadh.
Mossman T (1983). Rapid colorimetric assay for cellular growth and survival: an
application to proliferation and cytotoxicity assays. J Immunol Methods 65:
55 – 63.
Mousumi D, Malik CP, Bisen PS (2006). Micropropagation: A Tool for the
Production of High Quality Plant based Medicines. Current Pharm Biotech
7: 33 – 49.
Mukerjee T, Bhalla N, Singh AG, Jain HC (1984). Herbal drugs for urinary stones.
Indian Drugs 21: 224 – 228.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays
with tobacco tissue cultures. Physiol Planta 15: 473 – 497.
Murch S, Krishna J, Raj S, Saxena PK (2000). Tryptophan is a precursor
for melatonin and serotonin biosynthesis in vitro regenerated St. John's
wort (Hypericum perforatum L. cv. Anthos) plants. Plant Cell Rep 19:
698 – 704.
Murugai P (1959). In vitro culture of the inflorescence, flowers and ovaries of an
apomitic Aerva tomentosa Forest. Nature (Lond) 184: 72 – 73.
Murugan M, Mohan VR (2014). Phytochemical, FT-IR and antibacterial activity of
whole plant extract of Aerva lanata (L.) Juss. Ex. Schult. J Medicinal Plants
Studies 2: 51 – 57.
Muthu C, Ayyanar M, Raja N, Ignacimuthu S (2006). Medicinal plants used by
traditional healers in Kancheepuram District of Tamil Nadu. Ind J Ethnobio
Ethnomed 2:43.
Muthukumaran P, Shanmuganathan P, Malathi C (2011). Antioxidative and
antimicrobial study of Aerva Lanata. Asian J Biochemical Pharma
Res 1: 265 – 271.
Natarajan D, Shivakumar MS, Srinivasan R (2010). Antibacterial activity
of leaf extracts of Biophytum sensitivum (L.) DC. J Pharm Sci Res 2:
717 – 720.
xxii
National Committee for Clinical Laboratory Standards (NCCLS) (1998). Reference
method for broth dilution antifungal susceptibility testing of yeasts. approved
standard. NCCLS document M27-A. Wayne, Pa.
Natural Health Products Directorate (NHPD) (2000). An exploration of current
issues in botanical quality: A discussion paper. Health products and food
branch Canada 21 – 26.
Ncube B, Ngunge VNP, Finnie FJ, Van Staden J (2011). A comparative study of
the antimicrobial and phytochemical properties between outdoor grown and
micropropagated Tulbaghia violacea Harv. Plants. J Ethnopharmacol. 134:
775 – 780.
Nevin KG, Vijayammal PL (2003). Effect of Aerva lanata on solid tumor induced
by DLA cells in mice. Fitoterapia 74: 578 – 582.
Nikiforova VJ, Kopka J, Tolstikov V, Fiehn O, Hopkins L, Hawkesford MJ, Hesse
H, Hoefgen R (2005). Systems Rebalancing of Metabolism in Response to
Sulfur Deprivation, as Revealed by Metabolome Analysis of Arabidopsis
Plants. Plant Physiol 138: 304 – 318.
Niyogi SK (2005). Shigellosis. J Microbiol 43:133 – 143.
Norman AW, Demel R A, Kruyff BDE, Geurts Van KWSM, Van Deenen LLM
(1972). Studies on the biological properties of polyene antibiotics:
Comparison of other polyenes WITH filipin in their ability to interact
specifically with sterol. Biochem Biophys Acta 29: 1 – 14.
Ohyama K, Shinohara H, Ohnishi MO, Matsubayashi Y (2009). A glycopeptide
regulating stem cell fate in Arabidopsis thaliana. Nature Chemical Biol 5:
578 – 580.
Oktay M, Gulcin I, Kufrevioglu OI (2003). Determination of in vitro antioxidant
activity of fennel (Foeniculum vulgare) seed extract. Lebensm Wiss Technol.
36: 263 – 271.
Oluk EA, Ali C, Yasa I, Capanlar S, Kirmizigul S (2013). Comparison of the
antimicrobial activity and essential oil content of wild and micropropagated
Origanum sipyleum L.: A medicinal herb native to Turkey. J Medicinal
Plants Res 7: 230 – 233.
23
Ozgen U, Kazaz C, Secen H, Calis I, Coskun M, Houghton PJ (2009). A novel
Naphtaquinone glycoside from Rubia peregrina L. Turk J Chem 33: 561 –
568.
Pandey K, Sharma PK Dudhe R (2010). Anticancer Activity of Parthenium
hysterophorus Linn and Oldenlandia corymbosa Lam by Srb Method.
Scientific reports 1: 325 – 331.
Panlilio AL, Culver DH, Gaynes RP, Banerjee S, Henderson TS, Tolson JS et al.,
(1992). Methicillin-resistant Staphylococcus aureus in U.S. hospitals, 1975-
1991. Infection Control Hospital Epidemiol 13: 582 – 586.
Parani M, Anand A, Parida A (1997). Application of RAPD finger printing in
selection of micropropagated plants of Piper longum for conservation.
Current Sci 73: 81–83.
Park MK Park JH, Kim NY, Shin YG, Choi YS, Lee JG, KimKH, Lee SK (1998).
Analysis of 13 phenolic compounds in Aloe species by high performance
liquid chromatography. Phytochem Anal 9: 186 – 191.
Patra JK, Gouda S, Sahoo SK, Thatoi HN (2012). Chromatography separation, 1H
NMR analysis and bioautography screening of methanol extract of
Excoecaria agallocha L. from Bhitarkanika, Orissa, India. Asian Pacific
J Trop Biomed 2: 50 – 56.
Patricia Costa, Sandra G, Valentao P, Paula B, Andrade, Coelho N, Romano A
(2012). Thymus lotocephalus wild plants and in vitro cultures produce
different profiles of phenolic compounds with antioxidant activity. Food
Chem 135: 1253 – 1260.
Payal Singh S, Tribhuwan S, Rekha V, Jayabaskaranb C (2013). Liquid
chromatography-mass spectrometry based profile of bioactive compounds of
Cucumis callosus. European J Experiment Biol 3: 316 – 326.
Periaswamy B, Maier L, Vishwakarma V, Slack E, Kremer M, Andrews HL,
McClelland M, Grant AJ, Suar M, Hardt WD (2012). Live Attenuated
S. typhimurium Vaccine with Improved Safety in Immuno-Compromised
Mice. Plos one 7: 1 – 9.
Perumal Samy R, Ignacimuthu S, Patric Raja D (1999). Preliminary screening of
ethnomedicinal plants from India. J Ethnopharmacol 66: 235 – 240.
24
Peter KV (2004). Handbook of herbs and spices. Vol III CRC Press, Wood head
Publishing Limited, Cambridge.
Pierik RLM (1987). In vitro Culture of Higher Plants. Martinus Nijhoff, Dordrecht.
Pillai PP, Sajan JS, Kuttapetty M, Kuttanappilly S, Panicker J, Apian S (2012). ISSR
analysis reveals high intra specific variation in Rauvolfia serpentina L. A
high value medicinal plant. Biochemical Systematics and Ecology 40:
192 – 197.
Pinto G, Santos C, Neves L, Araujo C (2002). Somatic embryogenesis and plant
regeneration in Eucalyptus globulus Labill. Plant Cell Rep 21: 208 – 213.
Pizzale L, Bortolomeazzi R, Vichi S, Uberegger E, Conte LS (2002). Antioxidant
activity of sage (Salvia officinalis and S. fruticosa) and oregano (Origanum
onites and O. indercedens) extracts related to their phenolic compound
content. J Sci Food Agri 82: 1645 – 1651.
Plastina P et al., (2012). Identification of bioactive constituents of Ziziphus jujube
fruit extracts exerting antiproliferative and apoptotic effects in human breast
cancer cells. J Ethnopharmacol 140: 325-332.
Popa G, Cornea CP, Ciuca M, Babeanu N, Popa O, Marin D (2010). Studies on
genetic diversity in Amaranthus species using the RAPD Markers. Fascicula
Biologie 2: 280 – 285.
Pourmorad F, Hosseinimehr SJ, Shahabimajd N (2006). Antioxidant activity, phenol
and flavonoid contents of some selected Iranian medicinal plants. Afr J
Biotech 5: 1142 – 1145.
Powell W, Morgante M, Andre C, Hanafey M, Vogel J, Tingey S Rafalski A (1996).
The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers
for gemplasm analysis. Mol Breed 2: 225 – 238.
Prabodh S, Padmini S, Gopalkrishna B (2012). Isolation and Characterization of
Polyphenolic Compound Quercitin from Phyllanthus emblica. Int J Pharm
Sci Res 1520 – 1522.
Prakasha K, Anand R, Kabbali B, Gangadaraiah PL (2013) Regeneration of plantlets
from leaf derived callus of Aerva lanata Juss: A medicinal plant. Asian J
Biotech 4: 143 – 146.
25
Prasanna R, Harish CC, Pichai R, Sakthisekaran D, Gunasekaran P (2009). Anti-
cancer effect of Cassia auriculata leaf extract in vitro through cell cycle
arrest and induction of apoptosis in human breast and larynx cancer cell
lines. Cell Biology International 33:127 – 134.
Prieto P, Pineda M, Aguilar M (1999). Spectophotometric quantitative of
antioxidant capacity through the formation of a phosphomolybdenum
complex: Specific application to the determination of vitamin E. Analytical
Biochem 269: 337 – 341.
Priya CL, Kumar G, Karthik L, Bhaskara Rao KV (2012). Phytochemical
composition and in vitro antioxidant activity of Achyranthes aspera Linn
(Amaranthaceae) leaf extracts. J Agri Tech 8: 143 – 156.
Priya D, Alka C (2012). In vitro shoot culture of Aerva lanata (L.) A. L. Juss An
important medicinal plant. Innovative J Med Health Sci 2: 44 – 46.
Proestos C, Chorianopoulos N, Nychas GJE, Komatis M (2005). RP – HPLC
analysis of the phenolic compounds of plant extracts investigation of their
antioxidant capacity and antimicrobial activity. J Agri Food Chem 53:
1190 – 1195.
Pulido R, Bravo L, Sauro Calixto R (2000). Antioxidant activity of dietary
polyphenols as determined by a modified ferric reducing/antioxidant power
assay. J Agri Food Chem 48: 3396 – 3402.
Purohit SS, Mathur SK (1999). Drugs in Biotechnology fundamentals and
applications. Maximillan publishers, India. 576.
Qasim Samejo M, Memon S, Bhanger MI, Khan KM (2013). Comparison of
chemical composition of Aerva javanica seed essential oils obtained by
different extraction methods. Pak J Pharm Sci 26: 757 – 760.
Qureshi R, Bhatti GR (2009). Folklore uses of Amaranthaceae family from Nara
desert, Pakistan. Pak J Bot 41: 1565 – 1572.
Rady MR, Naglaa M Nazif (2005). Rosmarinic acid content and RAPD analysis of
in vitro regenerated basil (Ocimum americanum) plants. Fitoterapia 76:
525 – 533.
26
Ragavendran P, Sophia D, Arul Raj C, Gopalakrishnan VK (2011a) Functional
group analysis of various extracts of Aerva lanata (l.,) by FT-IR spectrum.
Pharmacology online 1: 358 – 364.
Ragavendran P, Sophia D, Arul Raj C, Starlin T, Gopalakrishnan VK (2012).
Phytochemical screening, antioxidant activity of Aerva lanata (L) – an in
vitro study. Asian J Pharm Clinical Res 41: 77 – 81.
Rahim Malek M (2012). Genetic relationships among Achillea tenuifolia accessions
using molecular and morphological markers. Plant Omics J 5: 128 – 135
Rahul C, Parimelazhagan T, Saravanan S, Sajeesh T, Arunachalam K (2012).
Antioxidant and Anti-inflammatory potential of Monochoria vaginalis
(Burm. F.) C. Presl.: A wild edible plant. J Food Biochem 36: 421 – 431.
Rai MK, Asthana P, Jaiswal VS, Jaiswal U (2010). Biotechnological advances in
guava (Psidium guajava L.): recent developments and prospects for further
research. Trees Structure Function 24: 1 – 12.
Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan AK (2011). Developing stress
tolerant plants through in vitro selection – an overview of the recent
progress. Environ Exp Bot 71: 89 – 98.
Raihan O, Brishti A Entaz B, Forhadul I, Mominur R, Syed MT, Aslam HM (2012).
Antioxidant and anticancer effect of methanolic extract of Aerva lanata
Linn. Against Ehrlich Ascites Carcinoma (EAC) in vivo. Orient Pharm Exp
Med 12: 219 – 225.
Rajanna L, Nagaveni C, Ramakrishnan M (2011). In vitro shoot multiplication of a
seasonal and vulnearable medicinal plant Aerva lanata L. Ind J Bot 3: 255 –
259.
Rajesh R, Chitra K, Padma M Paarakh (2011). Aerva lanata (Linn.) Juss. Ex Schult.
An overview. Ind J Nat Products Res 2: 5 – 9.
Rajyalakshmi P, Venkatalaxmi K, Venkatalakshmamma K, Jyothsna Y, Devi KB,
Suneetha V (2001). Total carotenoid and beta-carotene contents of forest
green leafy vegetables consumed by Tribals of south India. Plant Foods Hum
Nutr. 56: 225 – 238.
xxvii
Ramya V, Dheena Dhayalan V, Umamaheswari S (2010). In vitro studies on
antibacterial activity and separation of active compounds of selected flower
extracts by HPTLC. J Chem Pharm Res 2: 86 – 91.
Rao SG (1985). Evaluation of an experimental model for studying urolithiasis effect
of Aerva lanata on urinary stones. Indian Drugs 22: 640 – 643.
Rao SR, Ravishankar GA (2002). Plant cell cultures: Chemical factories of
secondary metabolites. Biotech Adv 20: 101 – 53.
Ratanasanobon K, Seaton KA (2010). Development of in vitro plant regeneration of
Australian native wax flowers (Chamela ucium spp.) via somatic
embryogenesis. Plant Cell, Tissue Organ Cult 100: 59 – 64.
Rauha JP, Remes S, Heinonen M, Hopia A, Kahkonen M, Kujala T, Pihlaja K,
Vuorela H, Vuorela P (2000). Antimicrobial effects of Finnish plant extracts
containing flavonoids and other phenolic compounds. Int J Food Microbiol
56: 3 – 12.
Raveesha KA, Prashanth KB, Lokesh G (2012).Regeneration of plantlets from leaf
derived callus of Aerva lanata, Juss: A medicinal plant. Asian J Biotech 4:
143 – 146.
Re N, Pellegrini A, Proteggente A, Pannala M, Yang, Evans CR (1999). Antioxidant
activity applying an improved ABTS radial cation declourization assay. Free
Radicals Biol Med 26: 1231 – 1237.
Rehman, RU, Israr M, Srivastava PS, Bansal KC, Abdin MZ (2003). In vitro
regeneration of witloof chicory (Cichorium intybus L.) from leaf explants
and accumulation of esculin. In vitro Cell Dev Biol 39: 142 – 146.
Ricca SM Cutting (2004). Display of heterologous antigens on the Bacillus subtilis
spore coat using CotC as a fusion partner. Vaccine 22: 1177 – 1187.
Rohlf FJ (2000). NTSYS-PC Numerical Taxonomy and Multivariate Analysis
System: Manual Applied of Biostatistics. Exeter Software, Setauket, NY,
USA.
Ryan KJ, Ray CG (2004). Sherris Medical Microbiology (4th Ed.) McGraw Hill.
1– 9.
Sahu AR, Chandra Rath S, Panigrahi (2013). In vitro propagation of Aerva lanata
(L.) Juss. Ex. Schult. Through organogenesis. Ind J Biotech 12: 260 – 264.
28
Sajeesh T, Arunachalam K, Parimelazhagan T (2011). Antioxidant and antipyretic
studies on Pothos scandens L. Asian Pacific J Trop Med 4: 889 – 899.
Sakpakdeejaroen I, Arun I (2002). Cytotoxic compounds against Breast
Adenocarcinoma Cells (MCF-7) from Pikutbenjakulj. Health Res 23:
71 – 76.
Sallah S, Semelka RC, Wehbie R, Sallah W, Nguyen NP, Vos P (1999).
Hepatosplenic candidiasis in patients with acute leukaemia. British
J Haematology 106: 697 – 701.
Salvador MJ, Pereira PS, Franca S, Regina C et al., (2003). Comparative study of
antibacterial and antifungal activity of callus culture and adult plants
extracts from Alternanthera maritima (Amaranthaceae). Braz J Microbiol 34:
131 – 136.
Samarghandian S, Shabestari MM (2013). DNA fragmentation and apoptosis
induced by safranal in human prostate cancer cell line. Indian J Urology 29:
177 – 183.
Sambrook J, Russell DW (2001). Molecular Cloning: A Laboratory Manual. Third
edition. Cold Spring Harbor Laboratory Press, Plainview, NY.
Sanches NR, Cortez DAG, Schiavini MS, Nakamura CV, Dias Filho BP (2005). An
evaluation of antibacterial activities of Psidium guajava (L.). Braz Arch Biol
Tech Int J 48: 429 – 436.
Sangwan RS, Sangwan NS, Jain DC, Kumar S, Ranade SA. RAPD profile based
genetic characterization of chemotypic variants of Artemisia annua L.
Biochem Mol Biol Int 47: 935 – 944.
Santanen A, Simola LK (1992). Changes in polyamine metabolism during somatic
embryogenesis in Picea abies. J Plant Physiol 140: 475 – 480.
Santos PRV, Oliveira ACX, Tomassini TCB (1995). Control microbiogicode
productos. Fitoterapicos Rev Farm Bioquim 31: 35 – 38.
Sara M, Naheed R, Muhammad S, Muhammad A, Tayaba I, Abdul J (2013) New
acylated flavonoid glycosides from flowers of Aerva javanica. J Asian Nat
Prod Res 15: 708 – 716.
Saraf A (2010). Phytochemical and antimicrobial studies of medicinal plant Costus
speciosus (Koen.). E J Chem 7: 405 – 413.
29
Sarlangue J, Brissaud O Labreze C (2006). Clinical features of Pseudomonas
aeruginosa infections. Arch Pediat 1: 13 – 16.
Sasidharan S, Chen Y, Saravanan D, Sundram KM, Yoga L (2011). Extraction,
isolation and characterization of bioactive compounds from plants extracts
Afr J Tradit Complement Altern Med 8: 1 – 10.
Schmourla G, Filho RRM, Alviano CS, Costa SS (2005). Screening of antifungal
agents using ethanol precipitation and bioautograhy of medicinal and food
plants. J Ethnopharmacol 96: 563 – 568.
Sethi A, Sharma SA (2011). Antioxidant activity with Total Phenolic constituents
from Aerva tomentosa Forsk. Int J Pharm Bio Sci 2: 596 – 603.
Shalini S, Prema S (2012). Phytochemical screening and antimicrobial activity of
plant extracts for disease management. Int J Current Sci 209 – 218.
Sharif A, Ahmed E, Abdul Malik, Mukhtar U Hassan, Ali Munawar M, Aleeza F,
Saeed Ahmad N, Jamil Anwar, Muhammad A, Zaid M (2011). Antimicrobial
Constituents from Aerva javanica. J Chem Society Pakistan 33: 439 – 444.
Shariff N, Sudarshana MS, Umesha S Hariprasad P (2006). Antimicrobial activity of
Rauvolfia tetraphylla and Physalis minima leaf and callus extracts. Afr
J Biotech 5: 946 – 950.
Sharma DK, Nandini Sharma, Rajni J, Vinod KJ (2013). Pharmacological and
phytochemical properties of Amaranthus (Amaranthaceae). Indian J Plant
Sci 2: 52 – 57.
Sharmin SA, Alam MJ, Sheikh MMI, Zaman R, Khalekuzzaman M, Mondal SC,
Haque MA, Alam MF, Alam I (2013). Micropropagation and antimicrobial
activity of Curcuma aromatica Salis a threatened aromatic medicinal plant.
Turk J Biol 37: 698 – 708
Shasany AK, Aruna V, Darokar MP, Kalra A, Bahl JR, Bansal RP, Khanuja SPS
(2002). RAPD marking of three Pelargonium graveolens genotypes with
chemotypic differences in oil quality. J Med Aromatic Plant Sci 24:
729 – 732.
Sher H (2011). Ethnoecological evaluation of some medicinal and aromatic plants of
Kot Malakand Agency. Pak Sci Res Essays 6: 2164 – 2173.
30
Shikha Singh, Ananya K, Sujata M, Enketeswara S, Sanghamitra N (2011).
Evaluation of phytomedicinal yield potential and molecular profiling of
micropropagated and conventionally grown turmeric (Curcuma longa L.).
Plant Cell Tiss Org 104:263 – 269.
Shilpha J, Silambarasan T, Pandian SK, Ramesh M (2013). Assessment of genetic
diversity in Solanum trilobatum L., an important medicinal plant from South
India using RAPD and ISSR markers. Genetic Resources and Crop
Evolution 60: 807 – 818.
Shilpha J, Silambarasan T, Virgin Largia MJ, Ramesh M (2014). Improved in vitro
propagation, solasodine accumulation and assessment of clonal fidelity in
regenerants of Solanum trilobatum L. by flow cytometry and SPAR methods.
Plant Cell Tiss Org 117:125 – 129.
Shirwaikar A, Issac D, Malini S (2004). Effect of Aerva lanata on cisplatin and
gentamycin models of acute renal failure. J Ethnopharmacol 90: 81 – 86.
Siddhuraju P, Manian S (2007). The antioxidant activity and free radical scavenging
capacity of dietary phenolic extracts from horse gram (Macrotyloma
uniflorum (Lam.) Verdc.) seeds. Food Chem 105: 950 – 958.
Siddiqui MA, Ismail Z, Saidan NH (2011). Simultaneous determination of
secondary metabolites from Vinca rosea plant extractives by reverse phase
high performance liquid chromatography. Phcog Mag 7: 92 – 96.
Sidhu Y (2010). In vitro micropropagation of medicinal plants by tissue culture.
Plymouth Student Sci 4: 432 – 449.
Sikarwar RLS, Kaushik JP (1993). Folk medicines of the Morena district, Madhya
Pradesh, India. Ind J Pharm 31: 283 – 287.
Singh AK (2004). Endangered economic species of Indian desert. Genet Resour
Crop Evol 51: 371 – 380.
Singh G, Rawat P, Maurya R (2007). Constituents of Cissus quadrangularis. Nat
Prod Res 21: 522 – 528.
Singh J, Singh AK, Khanuja SPS (2003). Medicinal plants: India’s opportunities,
Pharma Bio World 1: 59 – 66.
Singh KV, Nallapareddy SR, Murray BE (2007). Importance of the ebp
(Enterocarditis and Biofilm-Associated Pilus) locus in the Pathogenesis of
31
Enterococcus faecalis Ascending Urinary Tract Infection. J Clin Dis 195:
1671 – 1677.
Sivaram L, Mukundan U (2003). In vitro culture studies on Stevia rebaudiana. In
vitro Cell Dev Biol 39: 520 – 523.
Siveen KS, Girija K (2011). Immunomodulatory and antitumor activity of Aerva
lanata ethanolic extract. Immunopharm. Immunotoxi 33: 423 – 432.
Slusarenko AJ, Longland AC, Whitehead IM (1989). A convenient sensitive and
rapid assay for antibacterial activity of phytoalexins. Bot Helv 99: 203 – 207.
Sneath PH, Sokal RR (1973). Numerical Taxonomy: The Principles and Practice of
Numerical Classifi cation. Freeman, San Francisco, CA, USA.
Soam PS, Tribhuwan S, Rekha V, Jayabaskaran C (2013). Liquid chromatography-
mass spectrometry based profile of bioactive compounds of
Cucumiscallosus. European J Experiment Biol 3: 316 – 326.
Sohn HY, Son KH, Kwon CS, Kwon GS, Kang SS (2004). Antimicrobial and
cytotoxic activity of 18 prenylated flavonoids isolated form medicinal plants:
Morus alba L. Morus mongolica Schneider, Broussnetia papyrifera (L.)
Vent, Sophora flavescens Ait and Echinosophora koreensis Nakai.
Phytomedicine, 11: 666 – 672.
Soliman M (2006). A Cytogenetical studies on Aerva javanica (Amaranthaceae).
Flora Mediterranea 16:333 – 339.
Soundararajan P, Mahesh R, Ramesh T, Beguum VH (2006). Effect of Aerva lanata
on calcium oxalate urolithiasis in rats. Ind J Exp Biol 44: 981 – 986.
Srinivas P, Ram Reddy S (2012). Screening for antibacterial principle and activity of
Aerva javanica (Burm .f) Juss. ex Schult. Asian Pacific J Trop Biomed 2:
838 – 845.
Srinivas Reddy K, Reddy VM (2009). Antihyperglycaemic activity of ethanol
extract of Aerva javanica leaves in Alloxan- induced diabetic mice. J Pharm
Res 2: 1259 – 1261.
Srivivan K, Reddy V (2008). Antimicrobial studies on the leaves of Aerva javanica.
J Pharma Allied Sci 5: 495 – 499.
Stajner D, Boris M. Popovi T, Dusica Sali T, Stajner M (2014). Comparative study
of antioxidant status in androgenic embryos of Aesculus hippocastanum and
xxxii
Aesculus flava. Scientific World J. 2014:1 – 7. http://dx.doi.org/10.1155/20
14/767392.
Sturm S, Stuppner H (2000). Analysis of cucurbitacins in medicinal plants by High
Pressure Liquid Chromatography – Mass Spectrometry. Phytochemical
Analysis 11: 121 – 127.
Sua X, Kong L, Li X, Chen X, Guo M, Zou H (2005). Screening and analysis of
bioactive compounds with biofingerprinting chromatogram analysis of
traditional Chinese medicines targeting DNA by microdialysis/HPLC. J
Chromatograph A. 1076:118 – 126.
Sudhir K, Prasad KV, Choudahry ML (2006). Detection of genetic variability
among chrysanthemum radiomutants using RAPD markers. Current Sci 90:
1108 – 1112.
Sugimoto N, Kiuchi F, Mikage M, Mori M, Mizukami H, Tsuda Y (1999). DNA
profiling of Acorus calamus chemo types differing in essential oil
composition. Biol Pharm Bull 22: 481 – 485.
Sukanya SL, Sudisha J, Hariprasad P, Niranjana SR, Prakash HS, Fathima SK
(2009). Antimicrobial activity of leaf extracts of Indian medicinal plants
against clinical and phytopathogenic bacteria. AfrJ Biotech 8: 6677 – 6682.
Sulaiman CT, Gopalakrishnan VK, Indira B (2012) Reverse Phase Liquid
Chromatography Coupled with Quadra pole- Time of Flight Mass
Spectrometry for the Characterisation of Phenolics from Acacia catechu
(L.f.) Willd. Int J Phytomed 403 – 408.
Suleiman A, Abdul MA, Choudhary MI, Mustafa G (2010). Volatile Constituents of
Aerial Parts of Euphorbia aellenii Rech and E. microsciadea Boiss. from
Iran. Iranian J Pharma Sci 6: 63 – 66.
Suman PSK, Ajit K, Shasany MP, Darokar, Sushil Kumar (1999). Rapid isolation of
DNA from dry and fresh samples of plants producing large amounts of
secondary metabolites and essential oils. Plant Mol Bio Report 17: 1 – 7.
Sun YF, Song CK, Viernstein H, Unger F, Liang ZS (2013). Apoptosis of human
breast cancer cells induced by microencapsulated betulinic acid from sour
jujube fruits through the mitochondria transduction pathway. Food Chem
138: 1998 – 2007.
33
Surya MS, Ashiq M, Jayachandran K (2012). In vitro production of Vanillin from
suspension culture of Aerva lanata (L.) Juss. Ex. Shultes. Ind J Life Sci 2:
9 – 15.
Tadhani MB, Patel VH, Subhash R (2007). In vitro antioxidant activities of Stevia
rebaudiana leaves and callus. J Food Composit Anal 20: 323 – 329.
Taware AS, Mukadam DS, Chavan AM, Taware SD (2010). Comparative studies of
in vitro and in vivo grown plants and callus of Stevia rebaudiana (Bertoni).
Int J Integrative Biol 9: 10 – 15.
Thangavel A, Senthilkumar B, AarthyA, Senbagam D and Sureshkumar M (2014).
Phytochemical screening, gas chromatography-mass spectrometry (GC-MS)
analysis of phytochemical constituents and anti-bacterial activity of Aerva
lanata (L.) leaves. Afr J Pharm Pharmacol. 8: 126 – 135.
Thiyagarajan M, Venkatachalam P (2012). Large scale in vitro propagation of Stevia
rebaudiana (Bert.) for commercial application: pharmaceutically important
and antidiabetic medicinal herb. Industrial Crop Prod 37: 111 – 117.
Tosetti F, Noonan DM, Albini A (2009). Metabolic regulation and redox activity as
mechanisms for angioprevention by dietary phytochemicals. Int J Cancer
125: 1997 – 2003.
Trease GE, Evans CW (1984). Pharmacognosy. 12th Edn Balliere Tindall, London
257.
Trease GE, Evans CW (1989). Pharmacognosy. 13th (ed). ELBS/Bailliere Tindall,
London 345 – 773.
Trivedi, PC (2006). Herbal medicine: Traditional practices (Ed); Aavishkar
Publishers, Jaipur 322.
Uddin SN, Akond MA, Mubassara S, Yesmin MN (2008). Antioxidant and
antibacterial activities of Trema cannabina. Middle East J Sci Res 3:
105 – 108.
Udupihille M, Jiffry MTM (1986). Diuretic effect of Aerva lanata with water,
normal saline and coriander as controls. Indian J Physiol Pharma 30:
91 – 97.
Umadevi M, Sampath Kumar KP, Debjit B, Duraivel S (2013). Traditionally Used
Anticancer Herbs In India. Journal of Medicinal Plants Studies 1: 56 – 74.
34
Vadawale AV, Arve DM, Dave AM (2006). In vitro flowering and rapid
propagation of Vitex negundo L – a medicinal plant. Ind J Biotech 5:
112 – 116.
Valsaraj R, Pushpangadan P, Smit UW, Andersen A, Nyman U (1997).
Antimicrobial screening of selected medicinal plants from India.
J Ethnopharmacol 58: 75 – 83.
Van Acker S, Van Den WJF, Bast F (1996). Structural aspects of antioxidant
activity of flavonoids. Free Rad Bio Med 20: 331 – 342.
Van Wyk BE, Wink M (2004). Medicinal plants of the world. An illustrated
scientific guide to important medicinal plants and their uses. Timber Press,
Portland, Oregon. 480.
Varutharaju K, Soundar Raju C, Thilip C, Aslam A, Shajahan A (2014). High
efficiency direct shoot organogenesis from leaf segments of Aerva lanata
(L.) Juss. Ex Schult by using thidiazuron. Scientific World J 2014: 1 – 6.
Vasil IK (1988). Progress in the regeneration and genetic manipulation of cereal
crops. Nature Bio/Technology 6: 397 – 402.
Venkatamana M (2008). Ethnobotanical and Ethnoveternary plants from Boath,
Adilabad District, Andra Pradesh, India. Ethnobot Leaflets 12: 391 – 400.
Vetrichelvan, T, Jegadeesan M, Senthil Palaniappan S, Murali NP, Sasikumar K
(2000). Diuretic and anti-inflammatory activities of Aerva lanata in rats. Ind
J Pharm Sci 62: 300 – 302.
Vieira RF, Grayer RJ, Paton A, Simon JE (2001). Genetic diversity of Ocimum
gratissimum L. based on volatile oil constituents, flavonoids and RAPD
markers. Biochem Syst Ecol 29: 287 – 304.
Vijaya Kumar D, Raghavan KV (2000). Novel chromatographic fingerprinting
method for standardization of single medicines and formulations. Indian
Institute of Chemical Technology, Hyderabad. WO 0246739-EP2 0000991
991-263397CSIR G01N30-88.
Vijayan MN, Barreto Ida, Dessai SDS, Silva DR, Rodrigues A (2010).
Antimicrobial activity of ten common herbs, commonly known as
‘Dashapushpam’ from Kerala, India. Afr J Microbiol Res 4: 2357 – 2362.
35
Wagner H, Blandt S, Zgainski EM (1984). Plant Drug Analysis. Spring-Verlag, New
York 320.
Walsh TJ, Dixon DM (1996). Spectrum of Mycoses. In: Baron's Medical
Microbiology (Baron S et al., eds.) (4th ed.). Univ of Texas Medical Branch.
Wayne C, Hsin Lin LY, Chang TM, Huang WY (2012). Identification of two
licorice species, Glycyrrhiza uralensis and Glycyrrhiza glabra, based on
separation and identification of their bioactive components. Food Chem 132:
2188 – 2193.
Westh H, Zinn CS, Rosdah VT, Sarisa (2004). An international multicenter study of
antimicrobial consumption and resistance in Staphylococcus aureus isolates
from 15 hospitals in 14 countries. Microbial Drug Res 10: 169 – 176.
WHO (1998). Regulatory Situation of Herbal Medicines: A Worldwide Review.
World Health Organization, Geneva.
William JDB, Timothy G (2006). Andrews diseases of the skin: Clinical
dermatology. Saunders Elsevier. 46.
Wojdylo A, Oszmianski J, Czemerys R (2007). Antioxidant activity and phenolic
compounds in 32 selected herbs. Food Chem 105: 940 – 949.
World Cancer Report (WCR) 2014. World Health Organization. 2014: 1.
Wylie AH, Morris RG, Smith AL, Dunlop D (1984). Chromatin cleavage in
apoptosis; association with condensed chromatin morphology and
dependence on macromolecular synthesis. J Pathol 142: 62 – 67.
Yadav JS, Rajam MV (1998). Temporal regulation of somatic embryogenesis by
adjusting cellular polyamine content in egg plant. Plant Physiol 116:
617 – 625.
Yamunadevi M, Wesely EG, Johnson M (2011). A Chromatographic study on the
Glycosides of Aerva lanata L. Chinese J Nat Med 9: 210 – 214.
Yamunadevi M, Wesely EG, Johnson M (2011a). Phytochemical studies on the
terpenoids of medicinally important plant Aerva lanata L. using HPTLC.
Asian Pacific J Trop Biomed 220-225.
Yamunadevi M, Wesely EG, Johnson M (2012). Chromatographic finger print
studies on saponins of Aerva lanata (L.) Juss. Ex Schultes by using HPTLC.
Int J Curr Pharm Res 4: 52 – 57.
36
Yamunadevi M, Wesely EG, Johnson M (2012a). Chromatographic Studies on the
Tannins of Aerva lanata (L.) Juss. Ex Schultes. J Pharm 2: 41 – 51.
Yamunadevi M, Wesely EG, Johnson MA, Anto AA, Vinnarasi J (2013). GC-MS
studies on methanolic extracts of Aerva lanata. Indo Am J Pharm Research
3: 2687 – 2717.
Yang F, Yang J, Zhang X et al., (2005). Genome dynamics and diversity of Shigella
species, the etiologic agents of bacillary dysentery. Nucleic Acids Res 33:
6445 – 6458.
Yaseen KM, Aliabas S, Kumar V, Rajkumar SH (2009). Recent advances in
medicinal plant biotechnology. Ind J Biotech 8: 9 – 12.
Yasuhara R, Ohta Y, Yuasa T, Kondo N, Hoang T, Addya S, Fortina P, Pacifici M
(2011). Roles of beta-ketanin signalling in phenotypic expression and
proliferation of articular cartilage superficial zone cells. Lab investing 91:
1739 – 1752.
Ye M, Han J, Chen H, Zheng J, Guo D (2007). Analysis of phenolic compounds in
rhubarbs using liquid chromatography coupled with electrospray ionization
mass spectrometry. J Am Soc Mass Spectrom 18: 82 – 91.
Zapesochnaya G (1992). Canthin-6-one and beta-carboline alkaloids from Aerva
lanata. J Planta Med 58: 192 – 196.
Zhao J, Wang J, Chen Y, Agarwal R (1999). Anti-tumor-promoting activity of a
polyphenolic fraction isolated from grape seeds in the mouse skin two –
stage initiation –promotion protocol and identification of procyanidin B5-3′-
gallate as the most effective antioxidant constituent. Carcinogen 20: 1737 –
1745.
Zheng W, Wang SY (2001). Antioxidant activity and phenolic compounds in
selected herbs. J Agric Food CheM 49: 5165 – 5170.
Zhishen J, Mengecheng T, Jianming W (1999). The determination of flavonoid
contents on mulberry and their scavenging effects on superoxide radical.
Food Chem 64: 555 – 559.