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N o v e m b e r 2 3 - 2 4 , 2 0 1 8 | B a n g k o k , T h a i l a n d
Plant Science Congress 2018
Journal of Agricultural Science and Botany
ISSN: 2591-7897 | Volume 2
OF EXCELLENCE
IN INTERNATIONAL
MEETINGS
alliedacademies.comYEARS
PLANT GENOMICS
AND PLANT SCIENCE
World Congress on
Manju Anand, J Agric Sci Bot 2018, Volume 2 | DOI: 10.4066/2591-7897-C2-005
IN VITRO
MASS CLONING OF
STEVIA
REBAUDIANA
AND EXTRACTION AND
QUANTIFICATION OF STEVIOSIDE BY
HIGH PERFORMANCE THIN LAYER
CHROMATOGRAPHY
Manju Anand
Amity Institute of Biotechnology, India
S
tevia rebaudiana
Bertoni (family Asteraceae) popularly known as “Sweet
leaf” is an important medicinal plant used for obesity, heart disease, den-
tal caries, as contraceptive and anticancerous agent. The leaves of
Stevia
are
the source of diterpene glycosides and among these stevioside is a high in-
tensity, non-caloric, high potency sweetener being 300 times sweeter than
sucrose. An efficient and reproducible
in vitro
protocol was established for
the mass cloning of this valuable plant followed by extraction and purifica-
tion of stevioside--the major secondary metabolite from micropropagated
plants and cell cultures using High performance thin layer chromatography
(HPTLC). It exhibited a high propensity of
de novo
adventitious shoot forma-
tion both directly from the leaf explants and indirectly through leaf callus on
variously augmented Murashige and Skoog’s medium. Individual shoots were
rooted on half strength basal MS medium and plantlets were acclimatized
and successfully established in the field. Extraction of stevioside from leaves
of micropropagated plants collected at different time intervals (3, 4, 5, 18, 30
months), callus and suspension cultures were achieved following solvent ex-
traction with petroleum ether, methanol, diethyl ether and butanol. The crude
extract was initially purified on glass TLC followed by its fine purification on
pre-coated silica gel 60 F254 plates by using High performance thin layer
chromatography scanned at 210 nm. The highest amount of stevioside was
obtained from thirty months old plants which yielded 94.9 μg/ml of stevioside
followed by 69.40 μg/ml and 44.37 μg/ml in suspension cultures harvested
at stationary phase and callus respectively.
Manju Anand has completed her PhD from Panjab Uni-
versity, Chandigarh, India. Presently she is a Professor in
Amity Institute of Biotechnology, Amity Institute of Bio-
technology, Haryana, India. A professional in Plant Bio-
technology specializing in Plant Tissue Culture, she has
rich experience of working on the micropropagation of
some economically important hardwood and softwood
trees and edible bamboos and ascertaining their clonal
fidelity through different molecular markers. Presently
she is working on the mass propagation of some valu-
able and endangered medicinal plants and extraction of
secondary metabolites from
in vitro
cultures and their
evaluation as therapeutic agents. She has nearly 40 pub-
lications in peer reviewed journals that have been cited
over 192 times and her publication H-index is 9.
manand@ggn.amity.eduBIOGRAPHY