ANNUAL REPORT - Department of Biotechnology

complete and will be distributed to the collaborating
partners for back-crossing. Majority of equipment at
all three centres have been acquired. At BCKV, the
varieties to be used in the back-cross have been
identified. Procedures for the award of contract for
the construction of poly-house for back-crossing and
resistance testing have been initiated. Tungro
incidences in several districts of West Bengal have
been studied. At TNAU, the construction of the
controlled field testing facility has been initiated. The
varieties for back-crossing have been selected. To
help in the resistance evaluation, E. coli cells,
expressing the RTBV and RTSV coat proteins as
fusion proteins were constructed at UDSC. These
have been transferred to TNAU centre, where
immunization is in progress.
Development and analysis of cotton transgenics
for resistance to insect pests (Bollworm
complex)
More than 300 independent transgenics lines in
cotton (Coker 310FR) carrying the cry1Ac gene for
attaining resistance to Helicoverpa armigera have
been developed. In most of the transgenics the
cry1Ac gene is under the control of the double
enhancer CaMV 35S promoter, while in some it is
driven by either the FMV double enhancer or the
MMV double enhancer promoters developed in the
laboratory. One of the problems observed in
developing these transgenics was that a large
number of the initial transformants showed abnormal
phenotype and would not set seeds under green
house conditions. Analysis of junction sequences by
Southern hybridizations revealed that only a small
population of the initial transgenics developed had a
single copy of transgene. Currently the group is in
the process of assessing insect resistance vis-à-vis
expression of the cry1ac protein in the BC1/T1
progeny of 22 of the T0 lines which have either one or
two copies of the transgene. These lines showed no
abnormality in their phenotype and set seeds
properly when grown in field under containment net
houses.
Improvement has also been made in the
transformation protocol of cotton which allows the
use of imidazolinone as a selection agent instead of
kanamycin by using a double mutant acetolactate
synthase gene as marker. These modifications have
simplified the earlier protocol developed and used in
our laboratory as the transgenic embryos could be
directly germinated on MSO medium and transferred
to pots without the need of grafting. Our preliminary
observations also show that a higher percentage of
transgenics have normal morphology when selected
on imidazolinone as compared to selections on
kanamycin. Thus, different promoters as well as
innovative modifications in the gene per se need to
be tested. Testing large number of such modifications
by developing transgenics in cotton is not easy in the
current scenario. In order to overcome this an
expression system for cotton has been developed
which allows to test expression levels of any
transgene cassette quickly.
Biofertilizers
With growing environmental concerns, the sole
dependence on chemical inputs based agriculture is
being replaced by integrated approach involving
conjunctive use of both organic and inorganic
sources. In this context, biofertilizers have been well
accepted as an economical, cost effective,
renewable and safe organic source of plant nutrients
to sustain crop productivity. Moreover, with recent
focus on organic/bio-dynamic farming, the demand
of biofertilizers is likely to grow at a much faster rate
than before. At this juncture, we must realize that
microbial inoculants are an 'ecological inputs' whose
efforts are 'subtle and not dramatic' like chemical
inputs. Hence, inoculation with good quality
inoculants is a must and should be treated as an
insurance against failure of nodulation. The shelf life
both in the storage and transit needs to be improved
with due consideration to various 'abiotic' stresses.
The quality oriented production and marketing
network will certainly make biofertilizers a viable
enterprise for ultimate customer satisfaction.
Keeping these in view, programmes on development
of liquid biofertilizers and biofertilizers based
Integrated Nutrient management packages for
plantation crops and medicinal plants have been
generated. In addition, biofertilizers strains
developed through transgenosis will be evaluated in
contained conditions.
The report will be a document to highlight the
importance and the genesis of the programme,
scientific hypotheses behind the main networks (e.g.
49 Research and Development