ANNUAL REPORT - Department of Biotechnology

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Signal Transduction In a study supported at JNU to study signal transduction machineries in plant under osmotic stress, complete physiological characterization of genotypes of Brassica campestris, B. nigra, B.oleracea, B. juncea, B. napus and B. carinata for their tolerance has been achieved under laboratory conditions. Primers have been designed to isolate the middle portion of the hybrid type Histidine kinase from B. juncea (CS52) which has resulted in cloning and sequencing of G80 bp fragment. Novel transcription factors have also been cloned from three B. juncea varieties specific to salinity tolerance. RNA has been isolated and primers were designed using Atmyb2 conserved domain. The PCR-product was amplified and cDNA is being cloned. An efficient plant regeneration and transformation system has been optimized in selected commercial cultivars of B. juncea. A study has been supported at ICGEB to clone and characterize stress responsive promoters from Brassica sps. Putative promoter regions for stress related genes like SoS2, Histidine kinase (HKI), Myb and MyC like factors have been cloned. The amplified fragments for putative promoters have been cloned, which show 50-80% homology. Further work is going on to assess the true stress inducibility of these promoter sequences. Scientists at NCPGR, New Delhi has been able to clone 8 members of mitogen activated protein kinase family (MAPK) from rice. From the 8 clones, four full length genes have been obtained. They have further been cloned in PGEX vector having GST protein fusion for heterologous expression in bacterial system. MAP kinase induced by heat has been identified and work is on to transform rice with different constructs.To understand the mechanism operative during early signaling events mediating auxin dependent abiotic stress tolerance in groundnut at Calcutta University, CDPK has been characterized from Arachis hypogea showing 88% homology to a stress responsive CDPK from Arabidopsis. Result indicated that Dephsphorylation of AhCPK2 is not a response to water loss per se but to a change of water potential. No such change was 77 noted in the presence of ABA treatments indicating the noted oscillation of phosphorylation of this kinase to be an upstream event. Based on in silico analysis, 46-stress responsive transcription factors were identified from Arabidopsis highly stress responsive functional genes have also been identified and their promoter analyzed. No significant difference was observed for presence of transcription binding sites of stress responsive transcription factors belonging to 10 families. Functional involvement of a light signaling component (ZBF1) in root development in Arabidopsis. ZBF1 (Z-box binding factor 1), which codes for a bHLH protein has recently been cloned at NCPGR, New Delhi. The function of A novel transcription factor of light signaling on root development has been investigated. Mutations in ZBF1 results in elongated roots as compared to wild type plants. At NBRI, the expression of three Ethylene Responsive Factors (ERFs) was found ten days after watering was stopped. Increased transcript accumulation was also observed after a cold shock of 4 hours for LeERF5 and LeERF6 while expression of all three ERFs was repressed after a heat shock at 42°C for one hour. In order to functionally analyse these genes, they were expressed in a bacterial expression system in the vector pET28. The proteins isolated are currently being analysed. Transgenic tomato plants that overexpress the three ERFs have also been raised and are being studied for morphological and physiological changes. Floral Development To understand how floral organ development and regulation of size & shape of floral organs and leaves take place at molecular level, studies have been conducted at IISc, Bangalore on Arabidopsis. 17,000 EMS-mutagenized M2-seeds were screened and based on phenotypes, mutants have been categorized. To check the DNA binding properties of wild type TCPY, consensus binding site of TCP4 was determined using random binding site selection Research and Development
(RBSS) assay. Study was supported at IISc, Bangalore on functional analysis of meiosis in Arabidopsis using candidate gene approach. So far generation of AML RNAi constructs has been completed. The corresponding portion of the OML5 gene has been PCR amplified, cloned, and authenticated. A 3.6 kb region of the AML3 has been amplified and construction of the pAML2::GUS fusion in the reporter gene vector pBI101 has been completed. Transformation of Arabidopsis with AML RNAi construct has been done and multiple T1 transgenic plants have been obtained. Detailed characterization of phenotypes in 70 T1 plants screened has been completed. In 11 out of 70 T1 plants sterility was observed due to defects in male and female gametogenesis. Host Pathogen Interaction Studies have been supported during the year on Host Pathogen Interaction concentrating on the following areas :- Molecular basis of determinants of host-pathogen specificity; Functional characterization of R/ Avr genes, Early events in pathogenesis initiation with respect to adhesion, penetration mechanisms, hydrolytic enzymes etc., Study of signal transduction cascade involved in early events both in host as well as pathogen, Gene expression profiling of plant(s) and pathogen(s) during pathogenesis, Innate immunity / non-host resistance; Systemic Acquired Resistance; Induced Systemic Resistance etc., Mechanism of Pathogenicity, Genetic diversity of important plant pathogens., Genome sequencing and functional genomics of important pathogens. These studies have been supported to understand and develop the complex interactions between plant pathogen and host plants. SOL Genome An international consortium of ten countries, including USA, Korea, China, UK, India, Netherlands, France, Japan, Spain and Italy aims at sequencing the euchromatic region of the tomatog genome under the auspices of “The International DBT Annual Report 2006-07 78 Tomato Sequencing Project”. India which is one of the partner has been assigned ~12 Mb euchromatic region of chromosome 5 for sequencing. The Indian initiative on tomato genome sequencing is being taken up by 3 centres with UDSC and NCPGR sequencing the short arm, and NRCPB sequencing the long arm. In all, 38 BAC clones, covering approximately 3.8 Mb regions have been mapped to chromosome 5. Status Clones selected Marker cM Phase I SL_MboI0050C14 Phase III LE_HBa0191B01 CT101 0 Phase II LE_HBa0106O06 C2-At1g60440 0 Phase II LE_HBa0051A13 T1252 3 Phase II SL_MboI0005B15 Phase III LE_HBa0261K11 C2-At1g60200 7 Phase I SL_EcoRI0086I08 Phase III LE_HBa0042B19 cLET-8-B23 10 Phase I LE_HBa0189E17 T0564 11 Phase II LE_HBa0179E24 cLED-8-G3 15.5 Phase III SL_MboI0037H06 Phase I SL_MboI0095J08 Phase II LE_HBa0074A13 Phase III LE_HBa0058L13 T1592 16 Phase II LE_HBa0195M17 Library LE_HBa0165L04 TG432 21 Phase II Phase II Phase III LE_HBa0051A18 LE_HBa0027B05 SL_EcoRI0053P22 BS4 22 Library SL_EcoRI0073I01 Phase I LE_HBa0169M21 T1360 73 Phase I Library LE_HBa0334K22 LE_HBa0138J03 cLEX-13-G5 T1746 79 84 Phase II LE_HBa0166A02 T1777 105 Phase II Library Phase II Library Phase II Phase II Library Library Phase II Library Phase II Library Phase II Library LE_HBa0040C21 LE_HBa0025A19 LE_HBa0131D04 LE_HBa0100I06 LE_HBa0006N20 LE_HBa0108A18 LE_HBa0019C24 LE_HBa0141A12 LE_HBa0239D11 LE_HBa0013H09 LE_HBa0251J13 LE_HBa0028M20 LE_HBa0245E05 LE_HBa0076P16 T1584 TG69 CT130 TG185 TG597 CT138 108 111 115 119 119 119 Telomeric Region Euchromatic Region Heterochromatic Region Centromeric Region Heterochromatic Region Euchromatic Region Telomeric Region Sequencing status of tomato chromosome 5 Annotation of fifteen BAC clones using FGENESH revealed a gene density of one gene per ~5 kb sequence. Some locus specific variations in gene density were also observed. Studies have also been supported to understand the molecular basis of root development in tomato. Root specific subtractive library from tomato using Suppression Subtraction Hybridization (SSH, Clontech) has been construccted. 34 root specific EST's have been generated. Another 500 clones are in the process of sequencing. Subtractive library for temporal specific genes (8-day root and 40-day root) have also been constructed and are further being screened. TILLING (Targeting induced local lesions in genome) is an advanced selective molecular breeding technology that allows improvement of crop plants for which the genome sequences are known or in progress. At University of Hyderabad, a study has been initiated where TILLING technology is being Short Arm Long Arm UDSC & NCPGR NRCPB
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ANNUAL REPORT 2006 - 2007 Departmen
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CONTENTS Chapter-1: An Overview 1 A
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Chapter-6: Biotechnology for Societ
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An Overview Biotechnology is a set
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In a project to identify, map and t
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Medicinal and Aromatic Plants Four
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wax related gene fragments having h
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esearch; appropriate regulatory mec
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Science or Food Science & Technolog
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and the National Research Centre Ca
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many sophisticated instrumentation
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Advisory Structure Standing Advisor
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Human Resource Development The Depa
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a rapidly advancing area and teache
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per year for a period of three year
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this fellowship and 9 students have
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Page 37 and 38: throughput molecular marker service
Page 39 and 40: plasmamembrane localisation of GFP-
Page 41 and 42: antibiotic biosynthesis in Streptom
Page 43 and 44: the reporting period. Out of these,
Page 45 and 46: in progress. Besides multiplication
Page 47 and 48: primers (Operon) under the series O
Page 49 and 50: a total of 326 cotyledonary node ex
Page 51 and 52: Magnaportha grisea, showed signific
Page 53 and 54: some diseases are the two most impo
Page 55 and 56: For utilization of Multiple Aleuron
Page 57 and 58: BGA transformations, MPS transforma
Page 59 and 60: development of root knot nematode,
Page 61 and 62: plant growth and development. Due t
Page 63 and 64: significant weed control at seedlin
Page 65 and 66: three rice accessions with enhanced
Page 67 and 68: oth adults and nymphs. First year m
Page 69 and 70: c-DNA probes were developed using a
Page 71 and 72: across all the 462 clones with allo
Page 73 and 74: R & D programme have also been supp
Page 75 and 76: Inauguration of Butterfly Park by S
Page 77 and 78: Germlasing of NAPs conversed in gen
Page 79 and 80: patchouli were found to be similar
Page 81 and 82: With a view to isolate the taxol bi
Page 83: attributed to the ribosome inactiva
Page 87 and 88: showed sign of spoilage even after
Page 89 and 90: ody hepatitis (IBH) in domestic fow
Page 91 and 92: FSH, IGF-1 and cumulus granulosa mo
Page 93 and 94: uffalo UTMP cDNA exhibited 94.5, 88
Page 95 and 96: Biosurfactant Work on screening of
Page 97 and 98: conotoxins along with the posttrans
Page 99 and 100: Oligonucleotide probe for monitorin
Page 101 and 102: similarity with the already reporte
Page 103 and 104: studies on expressed sequences of t
Page 105 and 106: factors. Scientists at Sree Chitra
Page 107 and 108: A study was implemented at NCCS, Pu
Page 109 and 110: Medical Biotechnology Health relate
Page 111 and 112: F S C # C e lls 1000 800 600 400 20
Page 113 and 114: the pathogenesis of JE. It has been
Page 115 and 116: Delhi. The protocol for 4-colour wh
Page 117 and 118: Cross Sectional Morphology of the T
Page 119 and 120: Virus Research Centers or Networks
Page 121 and 122: As the clinical trials ae planned t
Page 123 and 124: een implemented for both basic and
Page 125 and 126: low cost for rapid diagnosis of dis
Page 127 and 128: Under the multi-centric project on
Page 129 and 130: The genetic basis of Type2 diabetes
Page 131 and 132: Floor model RBC Reactor - Close-up
Page 133 and 134: analyzed for the selection of best
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yield of hydrogen in the first stag
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Operational guidelines for collecti
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Programmes for SC and ST Population
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section in the Western Dun Valley t
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aromatic grasses. These women are g
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Poultry and Livestock Farming In an
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conducted on mushroom cultivation,
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probiotic selected strains of Lacto
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hydratase activity was shown by eig
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At Birla Institute for Scientific R
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study at Osmania University and IIC
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DBT Annual Report 2006-07 152
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In the area of recombinant pharma s
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MEC met five times during the year
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Biotechnology Information System Ne
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Bioinformatics National Certificati
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Meetings 1) International Conferenc
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Biotechnology Parks and Incubators
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International Collaboration Interna
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elow: Contraceptive and Reproductiv
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USP method. These included 59 smear
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Danforth); others (IHBT & possibly
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Memorandum of Agreement between Dep
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cytokine-mediated regulation of int
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In stem cell research, the studies
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two pathogens in the acidic environ
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software developed by TCS with supp
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eleased from activated microglia ar
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Genetic linkage mapping in Catharan
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three of the BACs (374Kb) of chromo
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ATPase mRNA binding protein, cathep
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Public Sector Undertaking Bharat Im
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complete protection against a paras
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Administration and Finance Administ
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2. As the Department wants the comm
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GENDER BUDGETING CELL The Departmen
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6. Prof. K. Dharmalingam Member Hea
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19. Prof. Mahtab S. Bamji Member Em
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INTER-DISCIPLINARY RESEARCH COMMITT
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MONITORING-CUM-EVALUATION COMMITTEE
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Dr. Debi P. Sarkar, Professor & Hea
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TASK FORCE ON BIOTECHNOLOGY BASED P
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TASK FORCE ON AGRICULTURAL BIOTECHN
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Prof. H. Sharat Chandra, Director,
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NATIONAL BIORESOURCE DEVELOPMENT BO
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Scientist 'F' Forest Research Insti
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Prof. B. N. Johri Department of Bio
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Annexure-III DBT SPONSORED UNIVERSI
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16. University of Jammu, 10 JNU-CET
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32. Visva-Bharati, 10 JNU-CET Prof
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46. Banaras Hindu 12 JNU-CET Prof.
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LONG TERM 237 Annexure-IV BIOTECHNO
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239 Annexures
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241 Annexures
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243 Annexures
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245 Annexures
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SOFTWARE AVAILABLE WITH BTISNet CEN
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249 Annexures
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12. Jagadish N, Rana R, Mishra D, G
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42. Vats D, Vishwakarma RA, Bhattac
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15. Singh, S., Upadhyay, A.K., Ajay
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PATENTS FILED / SEALED Dr. V. P. Ka
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22. Johny S, Kanginakudru S, Murali
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11. Mallappa, C., Yadav, V., Negi,
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Annexure-X STATEMENT OF BUDGET ESTI
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GST H2O2 HAU HCH hck HCMI HDAC1 HIC
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SATB1 SDL SFC SGOT SGPGI SGPT SH Si
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ANNUAL REPORT - Department of Biotechnology

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