ANNUAL REPORT - Department of Biotechnology

QTL for PHST) were also identified for eventual use
in fine mapping of the QTLs. Significant progress has
already been achieved in transfer of major QTLs for
high GPC and PHST into 10 elite Indian bread wheat
genotypes using marker-assisted selection (MAS)
during three backcrosses already completed; BC3F 1
generation will be raised in the coming rabi-season
and success will be evaluated again. The RILs for
bread quality were developed at DWR Karnal taking
HI 977 and CPAN 3004 as good bread quality parents
and HD 2329 as poor bread quality parent (in the first
phase of the project during 1995-2000). These along
with parents were grown at different agro-climatic
conditions of North Western Plains Zone, Central
Zone and Peninsular Zone. The RILs of both the
crosses were analysed for various quality
parameters like protein content, moisture content,
hectolitre weight, sedimentation value, thousand
grain weight including bread loaf volume and bread
quality score.
Salinity and dehydration stress tolerance in rice:
cloning of responsive genes, their promoters and
development of transgenics
Towards cloning of stress responsive genes, several
genes have been cloned. Amongst them one of the
gene encoding fructose 1,6 bisphosphate was
cloned to full length from Portresia (PCFR) and this
enzyme was found to be active in the presence of
NaCl. This gene is being functionally validated.
Another novel gene encoding glycine rich RNA
binding protein (GR-RBP5) has been cloned. It has
been shown that GR-RBP5 is nuclear-localized but
under stress conditions, bulk of this protein appears
in cytoplasm. On cloning of stress responsive
promoters and regulatory factors through 'genome
walking PCR' technique, promoters for various genes
that gets induced 'early' or 'late' with the persistence
of stress, and also which are down regulated have
been cloned. Using the 'promoter deletion approach',
the stress responsiveness of these promoters is
being validated by analyzing the reporter gene
activity in the transgenic system. The specific
transacting factors that bind to the specific ciselements
of the promoter have been cloned. The
OsBZ8 (an ABRE binding factor) has been shown to
be highly expressed in salt tolerant cultivars than in
salt sensitive cultivars of indica rice. Another
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transcription factor DREB2A (a DRE binding factor) is
being functionally validated using the transgenic
approach by over- and under-expression in indica
rice . Regarding genetic transformation of rice, the
PgNHX1 gene overexpressing transgenic rice
tolerates high level of NaCl and their seeds could also
germinate in the presence of salt. The glyoxalase II
gene also confers improved tolerance towards
salinity stress in transgenic rice.
Localization of OsGR-RBP5 homologous protein in tobacco BY2 cells. A.
a- Tobacco BY2 protoplasts (under uninduced control conditions) viewed
in bright field b- DAPI-stained protoplasts. c- protoplasts probed with anti
OsGR-RBP5 antibodies. B. a- Tobacco BY2 DaPI-stained protoplasts
o
(following 42 C, 1 h heat stress) b- protoplasts probed with anti OsGR-
RBP5 antibodies (arrowheads mark the nuclear regions) c-merged
image of panels a and b.
Relative salt tolerance of wild type (WT) and glyoxalase overexpressing
transgenic T1 generation rice plants grown in the continued presence of
200 mM NaCl. Note that WT plants could not sustain growth under this
condition while the transgenic was able to grow and set seeds.
Improvement of sorghum, finger millet and pearl
millet through application of biotech tools
Sorghum, pearl millet and finger millet are important
dual purpose crops in the arid and drier semi arid
tropics of north and north western and southern India
and are usually grown as rainfed crop. Drought and
some diseases are the two most important
Research and Development