ANNUAL REPORT - Department of Biotechnology

cloning vector and transformed in to E. coli DH5
competent cells. The expression of the toxin moiety
was examined through bioassay on Galleria
mellonella and Plutella xylostella. The LD50 value of
these cells was high compared to the original cells of
the strain indicating that the native cells of
Photorhabdus were more toxic compared to the
transformed cells of E. coli.
At University of Pune, molecular characterization of
antibiotic producing novel PGPR Acinetobacter
species is being done and so far 37 strains have been
isolated from rhizosphere of wheat and biotyped.
These strains were tested for their ability to inhibit the
growth of phytopathogenic bacteria and fungi. A. junii
A7 and A.haemolyticus A19 demonstrated strong
antibacterial and antifungal activity while
A.haemolyticus A22 demonstrated only anti-fungal
activity. Molecular identification of these three strains
was conducted using rDNA sequencing.
A.haemolyticus A19 produced antibiotic, which was
1
inducible by co-cultivation method and H-NMR
analysis revealed this antibiotic as pyrrolnitrin.
Characterization of other strains in order to optimize
antibiotic production is in progress.
At Lucknow University, studies on the
characterization of antiviral resistance inducing
protein gene of Clerodendrum inerme is being carried
out. CIP-29 protein was purified from the leaves of C.
inerme and polyclonal antibodies in rabbits were
produced. The anti CIP-29 serum was tested and it
was found to be specific. Cloning of the CIP-29 gene
was initiated with the designing of primers for RT-
PCR. One set of forward and reverse primers yielded
an approximate 950 bp length fragment, which is
being sequenced.
At TNAU, Coimbatore, a chimeric gene, cry 2Ax1 of
Bacillus thuringiensis was constructed by
engineering C-terminal region of domain-III in
Cry2Aa to improve toxicity of its protein against
Helicoverpa armigera. The constructed chimeric
cry2Ax1 gene was expressed in transformant of
acrystalliferous Bt strain. Analysis of Cry2Aa,
Cry2Ab, Cry2Ac and chimeric Cry2Ax1 proteins for
toxicity against larvae of H. armigera showed about
22-fold higher activity in the chimeric Cry2Ax1 protein
(in terms of LC ) than the Cry2Ab.
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At AMU, Aligarh, the concurrent antagonistic activity
of novel plant growth promoting bacterial strains,
which was isolated from rhizospheric soil has been
demonstrated against a range of phytopathogenic
fungi under in vitro conditions. Some of the strains
produce antibiotic Phenazine-1-Carboxylic Acid
(PCA). The PCA gene ~ 1.4 kb (phz C and D) has
been cloned and sequenced. The potential of strains
as super-bioinoculant for plant growth promotion with
parallel fungal growth suppression has been
established both in pot culture and field studies in
disease-conducive soil. Different combinations of
bioinoculants were tested and consortium No. 19,
consisting of P. aeruginosa - NJ15 + P. aeruginosa -
NJ101 + Rhizobium sp. -PS1 + P. putida - PS2 strains
in equal proportions has proved to be the most
efficient to increase the yield. Treatment with this
combination, significantly decreased the disease
incidence also.
At Pondicherry University, Pondicherry antagonistic
Fluorescent pseudomonad isolates viz. W5, Pf6, Pf11
and Pf13 were isolated. Strain, W5 exhibited a
broad-spectrum antifungal activity towards several
phytopathogenic fungi that attack rice, groundnut,
chilli, cotton, sugarcane, mango and banana.
Parthenium Weed Management
A major multicentric programme on the management
of Parthenium weed, which is an obnoxious weed,
was launched. Salient achievements of few centers
are as follows:
Control of Parthenium through eco-friendly approach
is being undertaken at NBRI, Lucknow. Effect of three
tree species Jatropha curcas, Terminalia arjuna and
Riccinus communis on the growth and control of
Parthenium was studied and R. communis was found
to suppress the growth of Parthenium maximally. The
active fractions of the different parts of tree species
were tested on seed germination. Bark fraction of T.
arjuna at only 2.0% concentration inhibit 100%
germination of seed. Leaf and bark extracted fraction
of T. arjuna was developed and tested on P.
hysterophorus and found to be effective at different
stages.
At RDU, Jabalpur, various Mycoherbicide are being
developed for the control of Parthenium. Colletotrium
gleosporioides, C. dematium, Alternaria alternate,
Fusarium oxysporum and F. solani, have shown
55 Research and Development