throughput molecular marker service (SSR, DArT) to Indian researches towards crop improvement and will also organize training programmes to students / scientists within India.
Programme Support
The following five projects have been supported in 'Programme Support' mode during the year: (i) Studies on Human Genetic Disorders at Banaras Hindu University, Varanasi; (ii) Research on Micronutrient deficiencies (including essential amino acids and fatty acids in reproductive age women) at St. John's Research Institute and Medical College, Bangalore; (iii) Immunotherapy of Cancer and Leishmaniasis at National Centre for Cell Science, Pune; (iv) Translational Research on Transgenic rice at University of Calcutta, Kolkata; (v) Biotic Stress in plants: A concerted approach based on functional genomic, proteomic and transgenic techniques at Bose Institute, Kolkata. In addition, three individual R&D projects have also been supported during the year: (i) Identification of genes from Arabidopsis for engineering of apomixes at Centre for Cellular and Molecular Biology, Hyderabad (ii) Identification of new molecular targets for the development of anti-cancer agents at Indian Institute of Chemical Biology, Kolkata (iii) Therapy of infectious and chronic diseases: targeted gene delivery and long-term specific modulation of gene expression at All India Institute of Medical Sciences, New Delhi and University of Delhi, South Campus, New Delhi.
The salient achievements of the projects supported under the 'Programme Support' mode during the year 2005-06 are summarized below:
Recombinant clones expressing erythropoietin (EPO), and interferon gamma (IFN-G) have been constructed and an expression clone for interleukin-2 (IL-2) is being designed toward developing technologies for therapeutic proteins at Institute of Microbial Technology, Chandigarh. Current work is also focusing on optimization of protein yields and folding for wild-type chains of all these three proteins from E. coli. Synthesis of the constructs coding for therapeutic monoclonal antibodies to be used for
assembling the framework for human antibody light and heavy chain genes is underway. Optimization of various assays crucial for testing antibody binding and neutralization of TNG-α and EGFr is in progress.
Putative homologous of genes encoding proteins of the soluble triglycerol biosynthetic complex (TBC) of Rhodotrula glutinis have been identified in Pichia pastoris at Indian Institute of Science, Bangalore.
Gene cloning, recombinant protein expression and biochemical characterization of these recombinant proteins is in progress. A single stranded DNA binding protein that binds to specific upstream sequences of alcohol oxidase (AOX 1) promoter of the methylotrophic yeast, Pichia pastrosis has been isolated and identified as zeta crystallin (ZTA1).
About 92 fungal isolates from garden soil and sugarcane baggase were screened for their Diacylglycerol (DAG) content. Four isolates showed a significant accumulation of DAG. The amount of DAG in one of the isolates was found to be very high (27% on dry weight basis). Several clones of M13 phage from the phage display library have been identified which bind specifically with either the hyphal or yeast form of Candida albicans. The clones have been sequenced and peptide sequences have been determined. Under the programme on cancer biology and therapeutics at the same institute, efforts have been initiated to identify gene expression signatures in various forms of breast cancers and breast cancer stem cells. About 96 breast cancer tissues and 82 normal breast tissue have been collected for microarray analysis. In order to elucidate the role of AP2 α in the progression of breast cancer, a total of 314 breast cancer cases (retrospective) alongwith the respective survival data of various clinical forms were collected.
Immunohistochemical staining for all these samples have been completed. Towards developing cancer cytotoxic drugs based on metal complexed compounds, two series of ruthenium and platinum complexes were prepared. The cytotoxicity of the complexes and the corresponding ligands has been established by MTT assay on cancer and normal cells.
The 'Programme Support' at M.S. Swaminathan
R e s e a r c h F o u n d a t i o n , C h e n n a i a i m s a t characterization and validation of the mangrove genes in the transgenic rice system for abiotic stress
tolerance. Fifteen putative transcription factor cDNA clones were identified from the sequenced Avicennia m a r i n a E S Ts . A g r o b a t e r i u m m e d i c a t e d transformation of potential transcription factors isolated from A. marina with constitutive and stress inducible promoters in rice system has been undertaken. One of the EST clones (Am 244) have been shown to be upregulated in roots of A. marina seedlings upon treatment with 500 m M of sodium chloride. Studies showed that Am 244 has a possible role in the response to salt stress in A. marina. Am 244 is found to be a putative plasma membrane protein and its transcript is expressed in both leaves and roots.
Plasmolysis with 0.8M Mannitol confirms
Expression studies with salt stress for the two transcription factors
Plasmolysis with 0.8M Mannitol confirms plasmamembrane localisation of GFP-PR244 in the transgenics
- Mannitol + Mannitol
0 6 12 24 48 12w 24w 0 6 12 24 48 12w 24w
Tr a n s g e n i c p l a n t s e x p re s s i n g P 5 C S ( p ro l i n e b i o s y n t h e t i c g e n e ) o n c o n s t i t u t i v e ( C a M V 3 5 S ) a n d i n d u c i b l e ( A B R E ) p ro m o t e r w e re d e v e l o p e d . E x p re s s i o n o f P 5 C S ( A ) , p ro l i n e a c c u m u l a t i o n ( B ) w a s s e e n o n l y u n d e r s t re s s w h e n e x p re s s e d o n i n d u c i b l e p ro m o t e r a n d t r a n s g e n i c s s h o w e d i m p ro v e d t o l e r a n c e t o d e s i c c a t i o n s t re s s ( C ) . C o n s t i t u t i v e l y e x p re s s e d p l a n t s s h o w e d d e c re a s e d g ro w t h u n d e r c o n t ro l c o n d i t i o n s ( C ) .
A E x p re s s i o n o f P 5 C S
W t - Wi l d t y p e I P - I n d u c i b l e p ro m o t e r C P - C o n s t i t u t i v e p ro m o t e r
N o n s t re s s St re s s C P I P
W T 1 2 3 4 W T 1 2 3 4
B . P ro l i n e l e v e l s i n w i l d t y p e a n d t r a n s g e n i c p l a n t s
N o n - s t re s s
St re s s
2 w e e k s a f t e r re c o v e r y Tr a n s g e n i c l i n e s ( I P ) W T
W T Tr a n s g e n i c l i n e s ( C P )
+ M a n n i t o l - M a n n i t o l
plasmamembrane localisation of GFP-PR244 in the transgenics
At University of Agricultural Sciences, Bangalore, work on transcriptome profiling of drought stress responsive genes from groundnut and their functional characterization has been initiated.
Among the genes isolated from Arachis hypogaea, nearly 70% had homologues in Arabidopsis thaliana as revealed by genome analysis. The functional relevance of some of the stress responsive genes was validated. A genomic library of groundnut was developed in lambda as a genomic resource for cloning genes of interest. A few full-length stress responsive transcription factors and functional genes have been cloned from both finger millet and groundnut for functional validation. Trangenics expressing DREB2A, DREB1A and NAC genes in groundnut and finger millet have been developed and are being evaluated for drought tolerance and preliminary studies showed stress adaptive response. Work on introgression of specific drought tolerance traits such as water use efficiency and root characteristics through molecular breeding to improve drought tolerance in rice has been initiated.
Biotech Facilities
The sophisticated biotech facilities have been set up in research Institutes / Universities spread across the country for wide spread use by scientists, institutions, industries and students engaged in biotechnology activities. The facilities undertake production and supply of Biologicals, reagents, microbial cultures and experimental animals. The facilities also conduct regular training programmes for capacity building in areas of instrumentations, handling of small animal houses, bioprocessing, microbial taxonomy and molecular Biology. The department has supported
establishment of following facilities during the year
2006-07:-i) Automated DNA Sequencing and Controlled Environment Plant Growth Chamber Facility at National Centre for Plant Genome Research, New Delhi.
ii) Creation of P3 facility for studying dangerous pathogens with special reference to anthrax causing pathogen Bacillus anthracis at Jawaharlal Nehru University, New Delhi.
iii) NMR facility for structure biology at National Institute of Immunology, New Delhi
The significant achievements at the ongoing facilities are as under:
International Depository Authority (IDA), Institute of Microbial Technology, Chandigarh:
The Microbial Type Culture Collection (MTCC) and gene bank supported during 1987 jointly by DBT and
st th th
CSIR became 1 IDA in India and 7 in Asia and 34 in the world in October, 2002. Besides IDA deposits, cultures for regular purposes and safe deposits are maintained in the IDA.
Repository for Filarial Parasites and Reagents at Mahatma Gandhi Institute of Medical Sciences (MGIMS), Sevagram, Maharashtra
The activities and achievements of the facility are as follows:
Maintenance of Brugia malayi infection in rodents (Mastomys & jirds): there are about 322 Mastomys and 285 jirds maintained in a CPCSEA registered animal house with about 24 Mastomys and 42 jirds carrying filarial infection. For breeding and rearing of Aedes aegypti mosquito colony (as vector for transmission of filarial infection to animal models), an Insectarium is also being maintained. Apart from being sources of parasite material & reagents, the animals have been used for filarial vaccine development studies involving Anna University, Chennai and BHU, Varanasi.
Maintenance of Filarial parasite Bank: presently there are about 40 million B. malayi mf, 500 male and female adult worms (from infected animals) and 1000 infective larvae (collected from infected mosquitoes) and about 2000 W.bancrofti mf collected from different endemic zones. The parasites from the bank from time to time have also been used for research work in the facility and supplied to VCRC (ICMR),
Programme support for priority areas in Plant Biotechnology at Bangalore University
The activities wise achievements are:
Mass propogation, establishment of germ plasm bank of rare, endangered and endemic orchids of Western Ghats: thirty different rare, endangered and endemic orchids species has been collected from Western Ghats. Out of them, 10 rare orchids were multiplied through in vitro seed germination.
Conservation, mass propogation, nursery technology and establishment of germplasm bank of rare medicinal plants of Sothern Western Ghats;
twenty five different rare critically endangered and high value target species of medicinal plants has been collected.
Ex situ conservation of forest genetic resources of Southern Western Ghats; the establishment of gene bank and collecting of species is in progress.
National Facility for Marine Cyanobacteria ( N F M C ) a t B h a r a t h i d a s a n U n i v e r s i t y, Tiruchirappalli
In NFMC, 400 marine cyanobacteria belonging to 25 genera and 65 species are maintained and available for research purposes. From the survey of Nicobar and Lakswadweep Islands, 110 and 112 strains of marine cyanobacteria have been recorded. From Palk Bay region, 50 marine anoxygenic photosynthetic bacteria are isolated and the identification is in progress. Identification tools have been developed with isoenzymes patterns, RAPD, 16sRNA , ITS, cpc and introns as markers. The facility has supplied 30 germplasm cultures to 30 Research Institutions and instrumentation facilities offered to 96 scholars outside the institution.
Centre for Genetic Engineering and Strain Manipulation at Madurai Kamaraj University, Madurai
The significant advances have been made with regard to study on M. leprae infection / reaction, regulation of chitinase expression and regulation of Pondicherry, Jiwajee Univ., Gwalior, Anna Uni.
Chennai, Dept. of Biotechnology, RTM Nagpur University, Nagpur, IIT Kharagpur & BHU, Varanasi.
Filarial serum bank: the bank has collection and storage of about 504 bancroftian filarial sera of different patient groups from different endemic zones (viz., Vidharbha, Raipur, Calicut, Mangalore, Bhubhaneshwar & Rourkela) and as well from non-endemic normal individuals. Sera from the bank have been supplied to BHU, Varanasi and Anna University, Chennai for diagnostic work.
Databases on diagnostics and management of filarial cases has been developed that provide information pertaining to region-wise clinical manifestations, filarial antigen and antibody positivity in different patient groups, duration of drug treatment requires for different clinical groups etc.
National Laboratory Animal Centre (NLAC) at Central Drug Research Institute, Lucknow
The National Laboratory Animal Centre being a prime source of research animals in the country to fulfill the requirements of animals to the out-side i n s t i t u t i o n s , u n i v e r s i t i e s a n d i n d u s t r i a l establishments for research purposes. During the year upto October, 2006, 140 cell lines has been supplied, 581 samples of fecal, blood, urine etc. of d i ff e r e n t a n i m a l s p e c i e s e x a m i n e d a n d microbiological monitoring was undertaken in respect of 835 samples. About 28000 animal lines have been supplied both within institution and to about 35 institutions both in public and private sector.
In addition, chest radiography and tuberculin testing services were rendered in 138 case and procuring &
utilizing 96 non human primates.
National Infrastructure Facility on Animal House at National Institute of Nutrition, Hyderabad
The center has been involved in breeding and supplying of different species and strains of animals all over the country. During the year, a total of 19139 animals were bred and 16341 animals were supplied. The center has also provided 510 ml of different blood products so far.
The equipment has been installed and the facility has been extensively used for teaching, training and research. Three training programmes have been conducted on software, maintenance and operations of flowcytometry and 20 scientists /students have been trained..
Upgradation of the existing computing infrastructure at the Bioinformatics facility at Centre for DNA Fingerprinting and Diagnostics (CDFD), Hyderabad
Sun Fire E20K High End Server and related software for hosting various tools and database received from EMBnet and APBionet has been installed for developing new tools and services for Bioinformatics and Biotechnology Research. The upgradation has a very positive impact on quality and quantity of bioinformatics related publications from CDFD.
Transgenic Green House Facilities at University of Agriculture Sciences, Bangalore and Tamil Nadu Agriculture University, Coimbatore
The green house with incinerator has been constructed with the objectives of testing of the transgenic crops already developed by these universities for evaluating their performance outside the laboratory, and to obtain Bio-safety data for large Scale cultivation and testing of the transgenic materials developed outside the Universities to test their performance and to obtain the Bio-safety data for large scale cultivation to derive their benefits.
Prof. S.P. Roychaudhari Drosophila Stock Centre at University of Culcutta, Kolkata
Drosophila stock centre maintains over 400 different genetic strains D. melanogaster. These stocks are used for teaching in various Universities and Institutes. Some of the strains has already been enlisted Drosophila Information Service, USA. 200 M.Sc. and Post M.Sc. students from different Departments of Universities are trained by the Scientists of the Centre. A workshop entitled
“Drosophila genetics: A new way to look teaching and Research resources” with 40 participants conducted.
antibiotic biosynthesis in Streptomyces. Major achievements include identification of biomarkers in serum of leprosy patients; identification of daunorubicin (anti-cancer antibiotic from Streptomyces) development of strain over producing chitinase; patenting of two skin genes; 6 international publication with impact factor 3 and above and t r a i n i n g o f 1 0 s t u d e n ts / s c i e n t i s ts f r o m industry/institution. The facility has developed technologies for production of chitinase, amylase and antibiotics.
DNA Sequencing Facility at University of Delhi, South Campus, New Delhi
The existing facilities have been optimally utilized and more than 1200 users are benefited. A 48 capillary automated DNA sequencer has been installed as phase III of the facility.
Toxicology facilities for GM Products at Shriram Institute for Industrial Research (SRI), New Delhi The Toxicology facility was inaugurated on 23.11.2006 and evaluations of GM products has already been started. Remodeling of existing animal house is completed and has been made functional.
Toxicological studies for DBT sponsored projects namely Natural Dye & Dunelliella Biomass has already been completed and Safety evaluation of the GM Tomatoes are in progress. These studies have been conducted on 50 % of the SRI's scheduled charges.
High precision microcalorimetry facility for m o n i t o r i n g b i o m o l e c u l a r s ta b i l i t y a n d interactions at Jawaharlal Nehru University, New Delhi
The main objective of the facility is to determine the thermal stability and conformational transitions of proteins, enzymes, nucleic acids and lipids in terms of transition temperature, enthalpy, free energy and heat capacity of denaturation. The equipments have been commissioned and are being utilized by various users.
Flowcytometry Facility at All India Institute of Medical Sciences, New Delhi
700 MHz NMR facilities for Biological Research (Jointly with DST) at IISc., Bangalore
The facilities has been utilized for structure studies on cytochorme b based fusion protein system, 5 Conotoxins, conformationally equilibrium forms of Holo-Acyl Carrier protein (PfACP) from Plasmodium falciparum. enzymes of the fatty acid biosynthesis pathway in the malarial parasite and Pantothenate Synthetase. During the year, two training programmes /seminars have been conducted with 68 participants and 42 resource persons. The facility has resulted in 37 publications in International journals.
Programme Support
Programme Support for Micropropagation Research and Technology Development
For large scale production and demonstration of tissue culture plants support continued to the two micropropagation technology parks and hardening units. A National Certification programme for insuring production of quality planting through tissue culture material was volved in consultation with Ministry of Agriculture. The salient achievements under the programme are given below:
Micropropagation Technology park at TERI, New Delhi
During the year, about 6.9 lakh tissue-cultured plants of various species were produced/ dispatched from DBT sponsored Micropropagation Technology Park at TERI. This includes forestry species (Paulownia, Dendrocalamus asper, D. strictus, Bambusa bambos, B. balcooa, etc), fruit crops (apple, citrus, strawberry), medicinal & aromatic plants (Aloe vera, Swertia chirata, Patchouli, Stevia, etc) and cash crops (Jatropha, hops). The tissue-cultured plants were distributed among various end users for field demonstrations and routine plantations. Production of a less-thorny type of Bambusa bambos, collected by Karnataka Forest officials was undertaken during the reporting period. More than 10,000 tissue cultured plants were dispatched from MTP. MTP shall be producing and dispatching approximately 15,000 plants of Bambusa bambos by July 2007. Mass
production of D. asper is in progress 10000, plants have been dispatched and it is aimed produce over 40,000 plants by end of August, 2007. In addition other species of Bamboo are also being produced B.
balcooa, B. nutans and Dendrocalamus hamiltonii, Mass multiplication of Anogeissus latifolia is being carried out for the industry. Presently, over 6,000 plants of this species are undergoing hardening in the nursery.
Approximately 4.90 lakhs of imported citrus rootstocks varieties, C-35, US-812, Benton, Carrizo, Rich 16.6, Troyer Citrange and Rangpur lime were dispatched to Punjab Agri Export Corporation during Feb. 2006 Sep. 2006. Fresh initiations of the varieties, which are in demand, were made and the cultures were sent for virus-indexing. The results have confirmed the virus-free nature of the mother
cultures. Currently, mass-production with freshly initiated rootstocks is underway as MTP is required to supply 2,00,000 plants in Feb-March 2007 and additional 2,00,000 plants in September-October 2007. Cultures have been initiated and plants being produced to meet target. In 2005, a new variety, 'Kamarosa' of strawberry was introduced into production. The plants of this variety were mass-produced and transplanted at TERI's Supi Centre in March 2006, for field trials and runner production. In addition to 'Kamarosa', 23,000 plants of two other varieties, Chandler and Ophra were produced during
Tissue culture raised Acacia mangium plants in field : 3 years old
the reporting period. Out of these, approximately 20,500 plants have been dispatched to various private growers. Approximately 55,800 plants of Aloe vera were dispatched to private growers during 2006.
Over 12,000 plants of a natural sweetener, Stevia rebaudiana were dispatched to various private growers in and around Delhi-Gurgaon region. Large-scale production of medicinal plant, Swertia chirata, was carried out and approximately 30,000 plants were dispatched. Approximately 4000 plants of 'Lahul Bitter' variety of Hops were supplied to a private group. This production was taken up as a contract production order.
Micropropagation Technology Park, NCL, Pune Extensive R&D was carried out on the micropropagation of identified crops species.
Following tree species were selected based on economic importance and requirement of user groups-Acacia mangium ,Casuarina equisetifolia, and Jatropha curcas. Extensive work on development and refinement of micropropagation protocols for economically important plants at tissue culture pilot plant, NCL, Pune have led to development of micropropagation technologies which have been tested for technocommercial feasibility by the way of large number of field verificatory trials, benefit to cost ratio analysis and technology transfer to industries. With a view to generate awareness about the superiority of tissue culture raised propagules among the farmers and user agencies, small scale demonstration plots using tissue culture propagules for horticulture crops and forest tree species have been established under this project. For horticulture species five demonstration plots were established using tissue culture raised p r o pa g u l e s o f b a n a n a , t u r m e r i c , g i n g e r, chlorophytum and patchouli. It was observed that tissue culture raised plants showed higher growth as compared to conventional plants. Growth of Banana var. mahan was best amongst the banana tissue culture raised plants. Two ratoon crops of these tissue culture raised banana were taken. The average yield was 30-40 kg/plant. The explants were taken from these plants for tissue culturing by a local
Following tree species were selected based on economic importance and requirement of user groups-Acacia mangium ,Casuarina equisetifolia, and Jatropha curcas. Extensive work on development and refinement of micropropagation protocols for economically important plants at tissue culture pilot plant, NCL, Pune have led to development of micropropagation technologies which have been tested for technocommercial feasibility by the way of large number of field verificatory trials, benefit to cost ratio analysis and technology transfer to industries. With a view to generate awareness about the superiority of tissue culture raised propagules among the farmers and user agencies, small scale demonstration plots using tissue culture propagules for horticulture crops and forest tree species have been established under this project. For horticulture species five demonstration plots were established using tissue culture raised p r o pa g u l e s o f b a n a n a , t u r m e r i c , g i n g e r, chlorophytum and patchouli. It was observed that tissue culture raised plants showed higher growth as compared to conventional plants. Growth of Banana var. mahan was best amongst the banana tissue culture raised plants. Two ratoon crops of these tissue culture raised banana were taken. The average yield was 30-40 kg/plant. The explants were taken from these plants for tissue culturing by a local