Of Withania Somnifera Dunal". Department of Botany Gulbarga University
Of Withania Somnifera Dunal". Department of Botany Gulbarga University
Parashuram.Y.J.
Biotechnology Division,
Bejo Sheetal Seeds, Pvt Ltd. Bejo Sheetal Circle
Mantha Road,
Jalna (MS)
431 203
[email protected]
[email protected]
Research
• Presently working as a Scientist at Biotechnology Division, Bejo Sheetal Seeds,
Pvt Ltd. Jalna (MS).
Work Profile
• Worked as a Research Fellow at Lal Bagh Plant Tissue Culture Lab, Hulimavu
Bangalore (October 1999- September 2000).
1
Publications:
6. High frequency plant regeneration from shoot tip culture of Chilli (Capsicum
annuum L.). Srinath Rao, G.S Pratibha, Y.J. Parashuram and C.P. Kaviraj
Plant cell Biotechnology & Molecular Biology. Volume 7 (3&4): 163-166.
2006.
2
Teaching
Conferences Attended:
3
8. International Conference on Plant Biotechnology and Molecular
Biology: 15-17 August 2008. Organized by Department of Biotechnology
Kakatiya University, Warangal- 506 009. A.P. India.
Educational Qualification
• B.Sc. with the combination of Botany, Zoology and Sericulture from the Gulbarga
University Gulbarga with First Class (61.4%) in 1997.
Computer Knowledge
• MS-Office (MS-world, MS-Excel, MS- Power Point)
• DTP (Desk Top Publishing)
• Good Knowledge of Internet Access
Personal Profile:
Name: Parashuram.Y.J.
Father’s Name Yallappa. J.
Date of Birth 09 June 1976
Sex Male
Marital Status Single
Nationality Indian
E-mail [email protected]
Optimized the media components and hormonal concentrations for the growth
of callus and cell cultures of Withania somnifera and Withaferin–A
4
accumulation. (Standardization of media components, use of different growth
regulators, collection of callus of different days, HPLC analysis for
Withaferin–A).
The ability of the in vitro raised shoots; roots and callus were assessed to
synthesize the Withaferin–A. (Effect of the individual growth regulator on
different explants and their effect on production Withaferin–A).
Initiated the Hairy Roots using two strains of Agrobacterium rhizogenes (LBA
4404 and LB 9402) and accumulation of Withaferin–A was studied. (Co
cultivation of different explants with Agrobacterium rhizogenes, induction of
hairy roots (Transformation), GUS expression, PCR and HPLC analysis for
Withaferin–A).
TECHNIQUES KNOWN
• Isolation of DNA
• Isolation of Plasmid
• Gene Transformation through Co-Cultivation and Particle
Bombardment
• GUS expression in transformed tissues
• PCR
• HPLC
5
“In vitro production of withanolides from cell and hairy root cultures of
Withania somnifera (L) Dunal”.
Introduction:
Tissue is an important area of Biotechnology can be used to improve the productivity of
planting material through enhanced availability of identified planting stock with desired
traits. A German scientist Haberlandt put forth the idea of cell and tissue culture in 1902.
All attempts to culture the plant cells and tissue culture were successful till the 1930.
Around 1939 White, Nobecourt and Gautheret independently reported the possibility of
culturing plant tissues for definite periods.
Micro-propagation is one of the important contributions of Plant Tissue Culture to
Medicinal plant propagation and has vast significance. The name micro-propagation
derives from the miniature shoots/plantlets initially produced from this method of plant
propagation. This technique provides a rapid reliable system for a production of large
number of genetically uniform plantlets. The primary goal of the tissue culture research is
crop improvement. Tissue culture techniques after advantages over the conventional
methods of plan breeding principally because new plant traits can be selected at the
culture level in the laboratory instead of the whole plant level in the field. This affords
advantages of time in that cellular screening for traits requires weeks or months where as
screening of whole plants typically requires entire growing season.
In recent time plant tissue culture in conjunction with Genetic engineering and related
techniques, is a promising and potentially emerging area of plant biotechnology and has
generated great interest and speculation for genetic manipulation of crop plants with
desirable results. The plant tissue culture was exploited both for basic and applied aspects
of plant research encompassing haploidy, mutagenesis, somatic embryogenesis,
somaclonal variations, protoplast fusion and genetic manipulation and even commercial
exploitation.
Plant description:
Withania somnifera (The Indian Ginseng) belongs to family Solanaceae includes 42
species and a common plant native of India. The plant is small subtropical medicinal
plant with anti carcinogenic and adaptogenic properties for which withanaloids and
6
sitoinlosides are responsible. It is a middle sized erect shrub, growing up to 1.5 meters
tall. It has egg-shaped, hairy leaves up to 10 cms, small, pale green flowers in clusters of
about 25 and smooth; spherical, red fruits with yellow seeds.
• Seeds were collected from the Agriculture Research Station Gulbarga, seeds
were germinated in vitro and different parts of the plants such as shoot tips,
cotyledons, shoots and leaves were used as explants.
• Callus induced from different explants such as shoots, leaf and root using
different concentrations of auxins viz., 2,4-D, 2,4,5-T, NAA and PAA alone
or in combination with BAP and Kn cytokinins.
• In an optimized media and explant, accumulation of withaferin-A was
measured.
• Development of single clones for high and stable production of secondary
metabolites (withaferin-A) in long-term culture by plating the cells.
• Multiple shoots were induced by using different explant viz., stem, leaf and
callus by using different growth regulators viz., BAP, TDZ and Zeatin alone
or in combination with IBA, Kn and IAA.
• Differentiate shoots, roots from callus culture and to asses their ability to
synthesize withaferin-A.
• In vitro regenerated shoots were transferred to rooting medium containing
different concentrations of NAA, IBA and PAA.
• Genetic transformation was achieved using two strains of Agrobacterium
rhizogenes.
• Initiation of hairy root culture using Agrobacterium rhizogenes and to know
their ability for the production of withaferin-A.
• The effect of various biotic elicitors (prepared from bacterial and fungal
culture) was carried if they can enhance withaferin-A accumulation.
• Co–cultivated tissue were examined for genetic transformation by GUS
analysis and PCR amplification.
7
• To immobilize the cells using polymers and to find out their role on
withaferin accumulation in cell culture.
• The production of withaferin-A was carried in a large-scale bioreactor using
free and immobilized cell culture.
Principal Investigator
Prof. Srinath Rao
Plant Tissue Culture and Genetic Engineering Lab
Department of Botany
Gulbarga University,
Gulbarga-585106
Karnataka- INDIA
[email protected]
+919986110109
References:
1. Dr.P.B.Kavikishore
Professor and Head 2. Dr. S.S Udikeri
Department of Genetics. Agriculture Research Station
Osmania University, Dharwad Farm (Hebballi Farm)
Hyderabad – INDIA Navalgund Road
[email protected] Dharwad – 7
+919948489567 [email protected]
+919448136821