ANNUAL REPORT - Department of Biotechnology

BGA transformations, MPS transformation of
PGPRs, Rhizobial transformations for rhizosphere
competence etc.) and the significant achievements
against completed projects supported with essential
information (abstracts of data, figures, diagrams
etc.).
Some of the projects under development of
transgenic biofertilisers programme have been able
to develop a few transformed strains of the
organisms. Further assessment of the scientific and
technological merits of the transformed strains and
prepare a plan of action for their testing for the
transformed properties by carrying out uniform
centralized trials in an institution are being carried
out. For the purpose, the identified centres who have
developed the strains will form a team incorporating
one member from the investigating UAS Dharwad.
The centres who have been identified to have
prospective transformed strains BARC for BGA,
University of Hyderabad for Azotobacter, UAS
Dharwad for Azospirillum and University of Baroda
for Pseudomonas. The action plan as to be prepared
by BARC in this regard will be finalized in the
beginning of 2007-2008 and one external expert will
be deputed to oversee and evaluate the conduct of
the trials and the results thereof.
Under a previous project JNU developed some
genetically engineered strains of Azotobacter.
Against an ad-hoc project under the network,
Division of Microbiology, IARI tested the performance
of these strains by container grown studies for 2
years. Results of the study have shown improved
growth promotional efficiency of these strains in
wheat. The strain is fit for further and larger testing
against a few more crops (e.g. rice) which will be
undertaken during kharif 2007 along with other
strains.
Significant scientific contributions has been made by
BARC in the development of strains of
Cyanobacteria. The notable achievements are:
standardization of electro-transformation protocol for
selected cyanobacterial strains, construction a novel
integrative expression vector (pFPN) for Anabaena,
cloning hetR and groESL in shuttle vector,
transformation, integration of pFPN and expression
from psbA1 promoter from Anabaena, cloning of
hetR, groESL and linA2 genes downstream to
promoter sequence in pFPN, construction of hetR
DBT Annual Report 2006-07
50
transformants of Anabaena sp. strain PCC7120 with
higher heterocyst frequency and nitrogenase activity,
construction of Anabaena sp. strain PCC7120
constitutively expressing groES-El with increased
thermotolerance.
The hyphal fusion of mycorhhiza programme, inter-
and intra-species hyphal fusion of the AMF by the
standardized protocol of co-culturing and injury repair
mechanism, characterization of the fusants
(progeny spores) employing morpho-taxonomic
markers, possible nuclei based recombinant
variation in physiological markers (glomalin
production, heavy metal accumulation) among the
fusants and some improvements in recombinant
strains with regard to such properties have been
achieved. The attempts to develop molecular
markers employing RFLP and marker based
distinction between parents and the presumptive
fusant strains will be undertaken during 2007.
The cloning of gdh gene and transformation of
Azotobacter strain for gdh mediated MPS phenotype
was successful. Results of in vitro studies showed
gain of MPS phenotype in the strains with
simultaneous reduction in nitrogen fixation ability.
UAS Dharwad centre could clone pqq synthase and
gadh genes from S. marcescens and K. pneumoniae,
transformed and expressed gadh gene in
Azosprillum and Pseudomonas. Demonstration of
MPS phenotype in the transformed strains and
results of plant growth promotion with a few of the
transformed strains was demonstrated.
MS University achieved in cloning and expression of
-
ppc gene under lac promoter in E.coli ppc mutant.
The system was used to successfully transform P.
fluorescens strains for over expression of the ppc
gene. The transformed strains showed high MPS
phenotype in buffered alkaline medium against TCP.
Qualitative and quantitative changes in organic acid
secretion profile of the transformed strains were
demonstrated. Effect of these strains on plant growth
promotion via P-solubilization could be studied
during 2007.
The intra-and inter-species molecular
phylogenetic differences among the widespread
fluorescent Pseudomonas species strains in plant
rhizospheres and development of molecular marker