ANNUAL REPORT - Department of Biotechnology

in progress. Besides multiplication of the elite winter
and spring types of doubled haploids, multi locational
evaluation was also conducted at different research
stations of the University encompassing varied
agroclimatic situations of HP including dry
temperate. Seven doubl haploids of different
maturity groups have been included in the Integrated
Disease Screening Nursery (IDSN) of the All India
Co-ordinated Wheat Improvement Project and
subjected for further evaluations. Another project
recently sanctioned at the same university is on
application of molecular cytogenetic approaches and
chromosome elimination techniques for genetic
upgradation of bread wheat and other hill crops for
various biotic and abiotic stresses. In the last couple
of months, various elite triticale, wheat lines
(hexaploid & tetraploid), wheat doubled haploids and
triticale x wheat derived recombinants along with
different maize lines (in the polyhouse) have been
raised in the fields for developing fresh triticale x
wheat hybrids and fix the already developed triticale x
wheat derived recombinants through maize and
Imperata cylindrica mediated haploid induction
techniques. Besides all efforts are being made to
establish and refine protocols for exercising in situ
hybridization in the Lab so as to utilize it further for
physical mapping of the alien gene introgressions in
bread wheat.
In a project on molecular analysis of dehydration
response in chickpea undertaken at NCPGR, New
Delhi interesting results reported. Plants suffer from
dehydration or water-stress and they respond by
inducing expressions a set of genes. Some of these
genes have been shown to provide tolerance to the
plants when expressed in high amount. In this project
towards screening for dehydration responsive
elements in chickpea, induction of expression of 101
genes under dehydration stress in chickpea
seedlings was carried out. One of the dehydration
inducible chickpea genes (CaCIPK) is a CBLinteracting
protein kinase (CIPK) homologue. CIPK is
a recently identified serine/threonine kinase family.
Computational analysis shows presence of a number
of putative stress-responsive elements in the
promoter. Different promoter deletion constructs
with GUS gene as reporter have been transferred in
tobacco.
Efforts were being made at University of Hyderabad,
DBT Annual Report 2006-07
38
Hyderabad, for development of transgenic
groundnut expressing synthetic cry IE-C and a rice
chitinase cDNA for insect resistance.The transgenic
plants reported earlier were characterized by RT-
PCR, southern and western analyses. Western
analysis on the transgenic plants using cry IC
antibodies showed that the synthetic cry I E-C protein
is expressed. Southern analysis using the PCR
amplified npt II fragment as a probe indicated two
integration sites on 3.5 kb and 6.0 kb fragments,
which are longer than the expected sizes indicating
integration of the transgenes into the groundnut
genome. Insect bioassay on the transgenic plant
progenies carrying the cry I E-C individually and in
combination with rice chitinase cDNA indicated that
the combination of genes had a better effect on insect
mortality, combination plants exhibiting faster
Spodoptera larval kill.
Under project in progress at CPRI recombinant
proteins of two putative subunits of potato RNase P,
Pop5 and Rpp25, were over expressed in
Escherichia coli and purified to homogeneity. RNase
P assay was standardized using reconstituted E. coli
RNase P protein (C5) and RNA (M1) subunits.
However, both the antisera immunoprecipitated
partially purified potato RNase P from floral buds.
Standardization of purification procedure of potato
RNase P to homogeneity and characterization of
subunit composition of the enzyme from two different
tissue sources are in progress.
The full-length and truncated versions of the BC1 and
BV1 genes have been amplified from the full-length
DNA B of the Madhya Pradesh isolate of the virus at
MKU Madurai. These have been cloned in the binary
vector pGA643 mobilized into Agrobacterium strain
KYRT1 and the high-yielding NRC37 variety of
soybean was transformed. PCR and Southern blots
indicated the presence of the inserted DeltaBC1
gene in the transformed soybean plantlets. Although
the wild type soybean could be regenerated from
embryonic tip explants, the transformed plants did
not form roots in the selection medium. Currently
work is in progress towards establishing all the
transgenic plants by micrografting. Molecular
analysis of the transgenic plants will then reveal the
presence and copy number of the transgenes.
Insecticidal crystal protein genes (cry) of Bacillus