ANNUAL REPORT - Department of Biotechnology

namely barley, wheat, rice, maize and Sorghum for
studying their conservation and cross-transferability.
For analyzing SNPS, twenty-two unigenes predicted
to be involved in sugar pathway were amplified in a
set of 13 sugarcane genotypes and sequenced. The
presence of SNPs was revealed in four genes
namely, sucrose synthase, sucrose phosphate
synthase, soluble acid invertase and sucrose
transporter. Polymorphism survey was extended to
25 genotypes belonging to different species,
varieties, related genera of sugarcane and five
cereals, and validated through CAPS analysis. 50
primers were designed for “R” gene analogues based
on sequences from NCBI database and Sugarcane
EST database (SUCEST). The chitinase gene (PR
protein) a major gene involved in defence
mechanism in crop plants, resistance protein (R30),
metallothionein like protein with possible
hypersensitive response regulation, receptor protein
kinase (RPK) involved in cell signaling, reversibly
glycosylated protein (RGP) involved in cell wall
biosynthesis and transcription factor (SSHR5) were
differentially expressed in cane tissues challenged
with C. falcatum. Coronatine insensitive gene (RcoiII)
involved in salicylic acid pathway regulation and
basal antifungal peptide (BAF) with antifungal activity
were the transcripts that were differentially
expressed in cell lines.
Molecular Characterization of Interspecific Hybrids of
Sugarcane Using Genomic In Situ Hybridization
(GISH) is being attempted at G.B. Pant University of
Agriculture & Technology, Pantnagar. The
chromosome count (utilizing the root tips) of 20
interspecific hybrids indicates that the somatic
chromosome numbers varies from 90 to 112. Some
of the interspecific hybrids were utilized in in situ
hybridization experiments using two repetitive DNA
clones as FISH probes. The physical localization of
these two clones is helping to develop FISH
landmarks on 10 to 14 pairs of chromosomes of these
interspecific hybrids.
Efforts are on to develop a diagnostic tool for
Sugarcane Grassy Shoot (SCGS) disease at ICGEB,
New Delhi and Vasantdata Sugarcane Institute,
Pune. A total of 339 samples with symptoms of
phytoplasma were collected from various fields of
Maharashtra, UP, Karnataka and Tamilnadu and
screened with a phytoplasma specific oligonucleotide
primers corresponding to two regions - 16SrRNA and
16S-23S rRNA intergenic region. Antibodies against
SecA and SecE proteins were developed and are
being used to screen the field samples at Vasantdada
Sugar Institute in order to develop the diagnostic tools
based on ELISA to detect the early stage of infection.
A PCR based diagnostic kit for red rot and smut
diseased of sugarcane has been developed at IISR,
Lucknow and is undergoing validation. It was
concluded that the designed primer pair is specific to
red rot pathogen and can amplify total genomic DNA
of wide variety of isolated or pathotypes causing
incipient infection in sugarcane stalks. Further to
validation of specificity of synthesized primer pair to
smut pathogen (Ustilago scitaminea) in three smut
affected varieties of sugarcane, infected stalks from
sugarcane varieties from Coimbatore, Pune and
Lucknow were sampled.
Over expression of Cry endotoxins from B.
thuringiensis in E.coli to test the efficacy against
sugarcane borers is being done at PAU, Ludhiana.
Cry 1Ac, Cry 1Aa3, Cry1IA5, Cry 1F were procured
from IARI, New Delhi. DNA of Cry 1Ac and Cry 1IA5
has been isolated and restriction analysis of
constructs has been done using different restriction
enzymes (Bam HI and Hind III). Sugarcane borers
collected from different places in the state are being
reared on natural diet in the laboratory for insect
bioassays using different Cry proteins.
Tea: Under a network programme on
“Characterization and improvement of tea through
biotechnological tools” involving IHBT, Palampur,
GBPHIED, Almora, Tocklai Experimental Station,
Jorhat, Bose Institute, Kolkata, University of Delhi,
Delhi, TERI, New Delhi, UPASI, Valparai, molecular
and morphological characterization has been
completed for 150 accessions. The study has been
extended to around 2000 accessions, including
garden selections. Genomic DNA has been isolated
from a total of 500 tea clones (350 of TRA, 100- IHBT,
and 50- UPASI). Pre-amplification was successfully
done in 462 tea clones by TERI, and was provided to
all the collaborative laboratories for running AFLP
gels. So far, AFLP profiles have been generated
63 Research and Development