ANNUAL REPORT - Department of Biotechnology

plant growth and development. Due to elicitors there
is more lignin accumulation which helps in
strenghtning the plant. Field trial data show that
average production of elictor treated field is 28
Kgs/plant and that of untreated field is 22 Kgs/plant.
Botanical Pesticides :
At North Maharasthra University, Jalagaon, studies
on the use of plant cyclotides are being conducted for
the management of storage pests. Few indigenous
plants belonging to family Rubiaceae and Violeaceae
were screened and bioactive molecules were
isolated, partially purified and characterized. The
isolated protein from Rubia cordifolia has shown
good antimicrobial activity in preliminary studies.
Pheromones & Semiochemicals :
In a collaborative project implemented at IICT,
Hyderabad and MPKV, Rahuri indigenously
synthesized pheromone components are being used
under Integrated Pest Management. The
pheromone blends of all the three species of
bollworms were synthesized using viable synthetic
routes with required purity and found to be
economical. Pheromone application technology was
demonstrated in cotton crop by mass trapping and
mating disruption. All the three species of bollworms
were effectively controlled by pheromone, which
confirms the efficacy of synthesized pheromone
blends. Pheromone dispensers were also developed
and compared with the imported ones for their
affordability and suitability and found to be
economical.
At IICT, Hyderabad, pheromones of Pomegranate
fruit borer and fruit sucking moths of sweet orange
were isolated, identified and synthesized. Several
synthetic pheromone standards ranging from C to
12
C carbon atoms both E & Z configuration were
16
screened against male antenna for their bio-activity.
Existence of female produced sex pheromone
communication was confirmed in different bio-assay.
Significant olfactory responses in male antenna
against female gland extract are recorded using
Electroantennogram (EAG). Studies on identification
of bioactive fractions of pheromone glands extract
are in progress. Preliminary GC-MS studies with
female glands extract have given valuable leads for
the characterization of bioactive fractions. Mass
DBT Annual Report 2006-07
54
rearing of Pomegranate fruit borer, Deudorix
isocrates successfully achieved at MPKV, Rahuri.
Molecular Studies on Biocontrol Agents :
Studies are underway to control sugarcane borer
through transgenic endophytic Acetobacter
diazotrophicus is being made at Pondicherry
University. Cry1Ac gene from Bacillus thuringiensis
was cloned using the shuttle vector pKT230, which
produce cry toxin protein of 130 kDa in recombinant
A. diazotrophicus. Preliminary bioassay studies
employing artificial medium to study the effect of wild
A. diazotrophicus and its transgenic and E. coli
containing Cry1Ac gene in the control of early shoot
borer revealed that the transgenic strains were able
to drastically reduce the live weight of neonate larvae
th
on 15 day post feeding but no larval mortality was
observed.
In a multicentric project being implemented at
ICGEB, and IARI, New Delhi, a 60 kDA native protein
of Xenorhabdus nemetophilus was purified from cell
lysate and oral toxicity was checked on H. armigera
and S. litura neonates. The protein has shown potent
oral toxicity on H. armigera with a LD50 value.
However, it was not potent against S. litura. The
recombinant 60Kda protein was also purified and
checked for oral toxicity on H. armigera however, no
oral toxicity on S. litura was recorded. A 4.5 kb DNA
fragment containing the GroEL homolog was
obtained by screening a genomic DNA library of X.
nematophilus. Analysis of the 4.5 kb DNA fragment
predicted an open reading frame of 1.7 kb, encoding
a 60 kDa protein. Primary sequence comparison of
the protein showed high homology with GroEL
protein of E. coli and Photorhabdus luminescence.
Gene specific primers were designed and a 1.7 kb
gene was amplified by PCR. The amplified DNA was
cloned for expression. The recombinant GroEL was
expressed as a soluble protein, which is being used
in the insecticidal assay
Molecular studies on insecticidal toxins of indigenous
P. luminescens and Xenorhabdus spp. are being
conducted at PDBC, Bangalore. Study confirmed the
presence of insecticidal toxin genes viz., TcdA1,
TcdB1, TcdB2, TccC1 and TccC3 in 6 isolates of
Photorhabdus luminescens and 4 isolates of
Xenorhabdus spp. Insecticidal toxin gene TcdB1 of
4.5 kbs from P. luminescens was cloned in to TA