ANNUAL REPORT - Department of Biotechnology

Studies on gene prospecting by comparative
proteomics in water stress legumes was supported at
NCPGR, New Delhi. The extracellular matrix (ECM)
proteome map of chickpea and the identification of
163 ECM-specific proteins has been completed. ESI-
MS/MS analysis has led to the identification of 134
drought related proteins (DRPs). Protein reference
map of nucleus in chickpea from purified nuclear
fraction has been developed. The largest percentage
of the identified proteins has been found to be
involved in signaling and gene regulation (36%),
while 17% are found to be involved in DNA replication
and transcription.
Under a study supported at Bharath Vidya Peeth,
Pune for enhanced Omega-3-Nutrition in Flax,
several primers based on the Omega-3 desaturase
were designed based on the sequences available.
Many primers amplified different loci and several of
these were cloned and sequenced. Two variants
each of delta-9 desaturase and Omega-3 desaturase
have been obtained. Omega 3-desaturase is being
cloned in binary expression vector for plant
transformation. The transformants are being
analysed.
Studies at TBGRI and RGCB, Trivandrum are
continuing on isolation and characterization of
gene(s) involved in the regulatory step(s) leading to
the biosynthesis of hypericin using transcript profiling
technology. Hypericin rich callus, shoot and cell
suspension cultures of Hypericum sp were
developed by optimization of nutrient media with
respect to PGR's and carbon source. Possible
enhancement in hypericin production through elicitor
treatment is in progress. Differential expression of
gene in NAA induced shoot and callus cultures is
being screened by Suppression Subtractive
Hybridization (SSH) analysis.
At IHBT, Palampur a cold tolerant gene has been
isolated and characterized from a plant of Lahul and
Spiti region. Transgenic Arabidopsis plants using the
Cu-Zn SOD gene have been developed and T1
generation has been obtained. Study of gene
behavior and its expression is under progress. Seeds
have been obtained from vacuum infiltered
Arabidopsis plants for beta helix-loop-helix (bHLH)
protein encoding transgenes. Selection process for
getting positive plant transformants is in progress.
Putative potato transformants for Cu-Zn SOD have
been obtained for two varieties using Agrobacterium
mediated transformation.
Bioactive Molecules
A Novel superoxide dismutase (SOD) enzyme was
isolated from Potentila and is being used for
preparation of antioxidant cream and other products
at IHBT, Palampur. A fully characterized heme
peroxidase from Acorus calamus with antifungal
activity has been purified at IFGTB, Coimbatore and
is being tested for antifungal activity against
Tricosporium vesiculosum. Further work on
validation of peroxidase on fungal hyphae is being
carried out in other plant species to understand its
function during defence. The toxic nature of the
enzyme which inhibited hyphal growth has been
demonstrated against phytopathogens such as
Macrophomina phaseolina, Fusarium moniliforme
and T. vesiculosum. This study indicates that
peroxidases may have a role to play in host defence
by inhibiting the hyphal extension of invading
pathogens.
Secondary metabolites from lichens sps. like
Rorcella montagnei, Dirinaria consimilis, Ramalina
pollinaria have been screened at MSSRF, Chennai
for presence of bioactive compounds. Two novel
compounds have been identified and toxicity testing
is being done.
Studies have been supported at RGCB and TBGRI,
Thiruvananthapuram on metabolic engineering of
andrographolide accumulation. Agrobacterium
mediated transformation protocol for Andrographis
paniculata has been developed and transgenic calli
overexpressing A.paniculata 1-deoxy-D-xylulose-5phosphate
synthase (DXS) gene apdxs1 have been
produced which on HPLC analysis show enhanced
accumulation of andrographolides as compared to
the control. Further work is on to develop a number of
primary transformants from the generating calli
transformed with apdxs1 and to do phytochemical
analysis of the transgenic Andrographis plants for
andrographoides.
In a study supported at TERI, New Delhi, biopesticide
formulation has been developed from plants of
Myrtaceae family. The formulation prepared has
been found to be effective not only against a
bollworm but was also against jassid population -
59 Research and Development