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RESEARCH HIGHLIGHTS
Major achievements:
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Micropropagation of important crop
plants, cash crops, ornamentals and forest &
horticultural trees,
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Development of transgenics in rice
chickpea, Brassica and cotton,
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DNA fingerprinting studies in
Basmati rice,
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Molecular mapping and tagging of
stress tolerant genes/ QTLs in rice and disease
resistant gene(s) in sorghum,
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Characterization of native B.
thuringiensis strains effective against
Helicoverpa armigera,
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Metabolic Engineering for higher starch
biosynthesis.
MICROPROPAGATION:
Initial success has been achieved in plant
regeneration in a few selected cotton cultivars (H
1098, H 1117, H 777, HS 6, RST9, RS875, Ankur651,
F846 and LHH144). Efforts are under way to improve
the efficiency of regeneration and subsequent
transfer to potted soil.
Regeneration has been successfully achieved
in five genotypes of wheat (HD 29, HD 2009, WH 157,
PBW 343, WH 533).
Micro-propagation of sugarcane varieties,
CoH92, CoH99, CoH101 and CoH110, was again carried
out using shoot tip explants and plants are being
maintained for the demonstration purpose.
Micropropagation of ornamental plant
species such as Chrysanthemum (Snowball,
Ghenghish Khan, Dignity, Kikubiori, Temptation and
Silk brocate), gladiolus and orchids was carried
out using the already-established procedure.
The cultures initiated from field
grown trees of female datepalm are being maintained
and multiplied. Some plantlets have been
transferred to experimental area on the farms. New
cultures from the offshoots obtained from field
grown trees have also been initiated for starting
fresh cultures. The work for maintenance of
cultures will continue as a demonstration of the
technique of in vitro multiplication of female
datepalms. The field transferred plants are being
monitored for the performance each year.
TRANSGENIC
RICE:
Transformation experiments were conducted to
transfer Potato protease inhibitor II (Pin2),
and barley late embryogenesis abundant protein (Lea3)
genes in Basmati rice varieties, Pusa Basmati 1 and
Taraori Basmati; A Japonica rice variety TNG67 was
used as the control. A. tumefaciens strains
containing useful genes (Pin2, Lea3)
driven by a suitable promoter (Pin2’,
Actin 1. In, ABRC) were used for the rice
transformation. Calli derived from mature seeds
and/or immature embryos were used. Over 200 hygR
plants have been obtained from these
Agrobacterium transformation experiments. About
40 plants have been transferred to the pots in the
transgenic greenhouse facility in 2000-2001. Further
molecular and progeny analyses of these plants are
in progress.
CHICKPEA:
Triparental mating has been done to mobilize
plasmids containing Bt and Pin II gene cassettes
into Agrobacterium strain LBA 4404. The
strains can now be used to transfer these genes into
chickpea varieties.
Medium requirements and culture conditions have been
standardized for plant regeneration from radicle
explant of embryo axis, immature cotyledons and leaf
explant of the in vitro grown seedlings & field
grown chickpea plants. Some success has been
achieved in transferring the regenerated plantlets
to potted soil.
Transient GUS expression at high frequency has been
obtained in embryo axis derived chickpea tissue
cocultivated with Agrobacterium strains
containing GUS gene as a marker.
DNA FINGERPRINTING
Laboratory facilities and protocols have
been developed for the DNA fingerprinting of rice
varieties using RAPD, AFLP, ISSR and microsatellite
(SSR) DNA markers.
A simple and cost-effective procedure has
been developed for the isolation of DNA from milled
rice samples, which contains higher amounts of
starch and RNA contents.
A DNA fingerprint database of 24 rice
varieties including commercially important Basmati
varieties, has already been developed using fifty
microsatellite DNA markers well distributed on the
12 rice chromosomes.
RAPD
markers proved highly successful in characterizing
the individual chickpea genotypes which are in
consistent with their pedigrees. Genotype H 98-107
was out grouped from other test genotypes as one of
its parents was a wild species, C. reticulatum.
However, screening with some more number of random
primers is required to characterize the chickpea
genotypes more accurately.
MOLECULAR MAPPING
SSR marker analysis was successfully used
to confirm the hybrid nature of F1 hybrids obtained
from Taraori Basmati x CSR10, Taraori Basmati x
Pokkali, Taraori Basmati x Azucena and Taraori
Basmati x New Plant Type II crosses.
Thirty SSR markers were screened for
Anthracnose resistance locus. SSR marker Xtxp61 and
Xtxp212 amplified a fragment of 550 bp and 700 bp,
respectively were found linked to the locus
conferring resistance.
SSR markers were more efficient that RAPD
markers as 63% of SSR markers differentiated between
the genotypes, whereas, only 39% of the RAPD markers
were able to differentiate the parental genotypes.
Experiments are in progress to develop the
Recombinant Inbred Line (RIL) populations from the
above four crosses. The material is in F3
generation.
By
screening with 198 random primers, 10 markers were
found linked to the locus for anthracnose in
sorghum. Two RAPD markers OPA 12 (apprrox. 0.5 kb)
and OPJ 01 (approx. 1.1 kb) very closely linked to
the locus were cloned.
Five RAPD markers were found linked to the locus
conferring resistance to leaf blight in sorghum. Two
markers, OPC 14 (approx. 0.6 kb) and OPH12 (approx.
2.1 kb), tightly linked to the locus were cloned.
Three RAPD markers were found closely linked to the
gene for resistance to oval leaf spot of sorghum.
VALUE ADDED-MICRO-ORGANISMS/ PRODUCTS
Efficacy of starch based Bt. Formulation
was evaluated under pot house condition on cotton.
Among the starch based formulations, revive
containing Bt. preparation was comparable to
commercial Bt. preparation against H.armigera
under pot house conditions.
Amylase gene of Bacillus sp.was cloned into
E. coli. Transgenic E.coli
was able to produce amylase on LB starch medium.
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