Annual Report 2011-2012 - Nabi.res.in

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTECONTENTSS.No Particulars Page No.1 From the Desk of the Executive Director 12. Vision and Mission Statement 33. Research Progress and Emerging Areas 54 Synergy Through Collaborations and Networking 415. Newly Joined Faculty 436. Research Publications 557. Extramural Grants and Funding 598. Governance 609. Human Resource 6910. Progress of Infrastructure at Main Campus 7311. Progress of Infrastructure at Interim Facility 7712. Important Events 8113. Photo-gallery 8514. Financials 98

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEFROM THE DESK OF THE EXECUTIVE DIRECTORDuring its second year of growth, NABI began to specialty moleculesprepare itself to function as a 'Change-Leader' for industry - thewho should anticipate the future needs of agri- starches, oils, proteinsfood sector, expand knowledge, and translate it and phytomolecules.to design food plants for the future. The A twenty first centuryGoverning Body constituted the first Scientific institute must generateAdvisory Committee (SAC) and the Programme daring ideas in bio-Advisory Committee (PAC) of NABI to guide its technology, beyondadvancement around turning points in agri-food the current GM technobiotechnology.Expanding the faculty and staff at logies and designNABI was the immediate need. The Programme plants for hybridAdvisory Committee in its first meeting held on vigour, photosynthetic potential, bioavailableJuly 26, 2011 emphasized upon establishing a nutrients, post harvest stability, specialtysystem for attracting bright young scholars. The products, nutrient and water utilizationGoverning Body approved a number of efficiency and endurance to biotic and abioticfellowships at NABI and stipend for short term stresses. Transformational inventiveness, teamtrainees. Soon, the institute developed critical work, big thinking, confidence to succeed frommass in some areas, and began to grow.farm to fork and societal priorities should set theThe faculty strength grew from seven last year toresearch agenda for NABI.twelve this year. The administration and Some of the important projects executed duringtechnical staff grew from seven to thirteen. the year included efforts towards theBesides faculty, four contractual scientists, identification of molecular traits and genes thatfifteen research fellows and seventeen trainees determine the processing and nutritional qualityenergized the institute and its resources. The in food crops, the ingredients that enhance theinfrastructure at the Interim Facility expanded in value of food for consumer health, the growthgenomics, tissue specific transcriptomics, and developmental pathways that determinemetabolomics, genome informatics, animal and seed formation and the quality of fruits,plant tissue culture, controlled environment transformational tools appropriate to designingplant growth facilities, food chemistry etc. An plants for future, and sustainable biologicalarchitect was appointed to design the NABI approaches for adding value to completecampus. Germplasm collection at experimental agricultural produce from seed to straw. Besidesfield became richer. The institute began to Arabidopsis as the model plant, the crop speciesvibrate with energy.in the above researches were wheat, millets,litchi, custard apple and mango. Details ofThe Governing Body and the Scientific Advisoryindividual projects are described in the chaptersCommittee inspired NABI to develop into athat follow.world class institute on the strength oftransformational innovations required for the The first research paper on the work 'conceivedshifting role of plants in bio-based economy. The and initiated' at NABI was accepted forcurrent objective of designing plants for publication. This was a motivating news in the'economic access to food' was shifting to second year of an institute that started fromdesigning plants differently for 'food sufficiency nothing. The paper reported the first finding of awith health and bioeconomy'. Some of the photo gemini virus infecting wheat in India. It wassynthetically most efficient plants will be published in the Archives of Virology. NABI willdesigned more creatively in future to produce hopefully use this virus to develop first1

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEindigenous tools in the form of gene silencing Programme Advisory Committees, Scientificand expression vectors for basic and applied Advisory Committee, Governing Body, thestudies on wheat. Discovery of the wheat virus in Society of NABI; and hard work of the facultytwo geographically and climatically distinct and staff. The heritage of a rich beginning shouldlocations (Mohali and Nilgiri) may be a pointer take the institute to greater accomplishments inat taking this new virus seriously by the breeders the years to come.and wheat pathologists in the country.It is a continuing journey for NABI. MyThe first sponsored research project that NABI colleagues have done well in preparing thereceived was in the important area of institute to match the high expectations ininformation flow from root to seed. The early transforming plant sciences. A new institute likefindings on the possibility of moving small RNA NABI is freer to sketch and plan its areas forsignals across the plant, can open valuable investment, when the resources and details areopportunities for regulating economically and still uncommitted, than at a later stage when theagronomically important traits. The applications die has been cast and one must live with what theof mobile small RNA signals can open past had chosen. Through this Annual Report, Iextraordinary possibilities for designing plants. urge the readers and well wishers to guide us toNABI should hopefully apply this technology to the path of exceptional creativity for addressingdevelop smarter horticultural plants that are the changing landscape and dynamics ofdifficult to improve due to long growing seasons. knowledge for national good and globalThe emphasis on commitment, creativity,networks.infrastructure, intellect and team work shouldkindle high ambitions in the young faculty andresearch fellows to keep NABI ahead of time inaddressing the challenges in nutrition, food(Dr Rakesh Tuli)security and biobusiness. A good beginning hadExecutive Directorbeen made - thanks to the insights andencouragement by the members of theNational Agri-Food Biotechnology Institute2

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEVISION AND MISSION STATEMENTTo be a nodal organization for knowledge generation and translational scienceleading to value added products based on agri- food biotech innovations.llTo transform agri-food sector into globally rewarding and sustainablebiotechnology-based enterprise through innovative solutions in primaryand secondary agriculture including high-end food processing.To develop synergy among knowledge providers and investors in agrifoodsector to carry innovations to marketplace.3

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTERESEARCH PROGRESS ANDEMERGING AREAS5

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEwheat by using synchrotron powered beam different tissues of mature wheat grain ofline-VESPERS (Very Sensitive Elemental contrasting genotypes at University ofand Structural Probe Employing Radiation), Ljubljana and Jozef Stefan Institute,at Canadian Light Source (CLS), Saskatoon, Ljubljana, Slovenia. The µ-PIXE imagingCanada. XRF images obtained of the crease analysis of Aegilops kotschyi acc. 3790region of diverse genotypes showed revealed the presence of high concentrationsdifferent iron localization pattern. The high of iron in crease region, in particulariron genotypes (Aegilops kotschyi acc. vascular strands (Figure 3). The other iron3790, Triticum aestivum L. IITR26) showed rich tissues were nucellar projection, therelatively high intensity of iron near crease-aleurone, aleurone and scutellum.pigment strand and nucellar projection The endosperm showed significantly lowtissues of crease region (Figure 2). On the iron accumulation. Its embryo regionother hand, in the low iron wheat genotypes showed only little increase in ironFigure 2: Iron XRF maps of transverse section of seed of (A) Aegilops kotschyi accession 3790. (B)Triticum aestivum L. ITR26. (C) Triticum aestivum cv. WH291. (D) Triticum aestivum cv. WL711. Thecolour scale represents relative intensities of fluorescence, with blue and red corresponding to the lowestand highest iron intensity, respectively.(Triticum aestivum cv. WH291, Triticum accumulation than in endosperm. Ironaestivum cv. WL711) the high intensity distribution analysis in Triticum aestivum L.region was mobilized towards aleurone of IITR26 showed nucellar projection as ironcrease (Figure 3). XRF scanning in all four hot spot, carrying highest concentration ofgenotypes showed aleurone as the iron hot iron. The next iron rich tissues were creasespotregion with very low signals for iron in aleurone, aleurone and scutellum. Inthe internal layers of endosperm (Figure 2).vascular strand, iron was three times lesserthan in nucellar projection, indicating2. µ-PIXE iron localization maps have beenmobilization of iron from vascular tissue togenerated to further validate the µ-XRF andnucellar projection.to get accurate quantification of iron in8

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEFigure 3: Iron µ-Proton induced X-ray emission (µ-PIXE) maps of cross-sectionof grain of (A) Aegilops kotschyi accession 3790. (B) Triticum aestivum L.IITR26. (C) Triticum aestivum cv. WH291. (D) Triticum aestivum cv. WL711.PIXE element maps of low iron genotypesTriticum aestivum cv. WH291 depictedcrease-aleurone, nucellar projection andaleurone as major tissues for accumulationand localization of iron, followed byscutellum and vascular strand. On the otherhand, in WL711, the genotype having lessgrain iron content, aleurone was the ironrichest tissue followed by nucellarprojection and scutellum. In nucellarprojection, iron concentration was half ofthe aleurone. In vascular tissue, iron wasthree times lesser than that in aleurone.Thus, in contrast to the high iron genotypes,a gradual increase in iron concentration wasobserved along the mineral-transport route(from vascular tissue, nucellar projection tocrease-aleurone) of both of the low irongenotypes. However, significantly lessrelative amount of iron in endosperm of allthe genotypes affirms aleurone as the majorhurdle in iron mobilization towardsendosperm.3. µ-XANES has been recorded from differenttissues of crease region of mature wheatgrains to identify different ionic forms ofiron and chemical environment around ironFigure 4: Laser captures micro-dissection of nucellar projection transfercells (NP), vascular bundle (VB), aleurone (AL) and endosperm transfercells (TC) of wheat grain at 14th days after anthesis.9

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEFigure 5: Electropherogram of RNA, extracted from seed tissues of WL711:VB=Vascular Bundle, NP= Nucellar Projection, E= Endosperm, Al= Aleurone,showing RIN value and RNA integrity assessed on Bio-analyser (Agilent).metal at CLS. XANES of bulk seeds are inprogress at Jozef Stefan Institute, Slovenia.Extended X-ray absorption fine structure(EXAFS; at Hamburg SynchrotronRadiation Laboratory, Hamburg, Germany.)will be performed on the mature wheat grainsamples to get the chemical form in whichiron is localized in the mature wheat grains.4. To examine the tissue specific differentialtranscriptome in the tissues of developinggrains of contrasting genotypes, LCM wasapplied to micro dissect tissues such asvascular bundle, nucellar projection,endosperm and aleurone (Figure 4). RNAextraction from the micro-dissected tissueshas been established. The RNA IntegrityNumber (RIN) was above 7 in each case(Figure 5), hence suitable for tissue specifictranscriptomics studies to be taken up in thefuture.1.1.2 Gene discovery for improvement ofprocessing and nutrition quality in wheatCo-ordinator:Rakesh TuliInvestigators:Joy K. RoyShrikant Subhash MantriProject Scientist:Nishima WangooResearch Fellows:Anuradha SinghMonica SharmaIntroduction:Wheat is one of the most important staple foodcrops and wheat flour is processed into a widerange of baked and processed foods. Althoughwheat breeders have been successful in achievinghigh yields, but the present high yielding varietiesrequire improvement in processing and nutritionrelated traits to meet the increasing demand ofhealthy wheat diets for consumers and betterprocessing quality for baking and processingindustries. Wheat is also a good source of dietaryfibres (germplasm & bran) i.e. non-starchcarbohydrates known to be helpful in theprevention of diet related health problems such asdiabetes, hypertension and some types of cancers.Better understanding of genes and regulators ofmetabolic pathways and their interaction withenvironment is required for the improvement ofseed quality for nutrition and processing.Research Objectives:lTranscriptome and small RNA sequencingand genomic expression studies using wheatmicroarrays to identify genes and regulators10

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEof starch and phenolics biosynthesis acid, cytokinin, ethylene and abscisic acid),pathways having significant effects on c e l l d i v i s i o n , h e a t s h o c k , c o l dprocessing and nutrition quality.acclimatization, storage proteins andtranscription factors were examined.ll Evaluation of effects of variation inintermediates and products of the above 3. Gene expression profiles for nicotianaminepathways on processing and nutrition synthase, ferritin, metallothionine andquality.phytic acid pathway genes were identified.l Validation of gene function.These can be potential candidates for theimprovement of wheat for highLong Term Objectives:accumulation and bioavailability ofmicronutrients such as iron and zinc. BasedTo improve the high yielding Indian wheaton sequence homology with the inventoryvarieties and/or advanced breeding lines fordatabase of wheat transcription factorsprocessing and nutritional quality to meet the(wDBTF), twenty three probe sets showingincreasing demand of healthy wheat diets for>10 fold changes at least at oneconsumers and better processing quality fordevelopmental stage were identified.baking and processing industries.Among them, two transcription factors:Research in Progress:Plant Homeo Domain (PHD) and C2C21. Four popular varieties of Indian wheatGATA showed 50 fold change (Figure 6).(C306, Lok1, Sonalika, WH291) were 4. Peroxidase and some homologs were upevaluated for processing and nutrition traits.regulated 30 fold at 28 DAA, while2. Genomic expression was compared at three granule bound starch synthase (mutantdevelopmental stages by using Affymetrix GBSS1, Ta.24114.7.A1_at), coldwheat arrays. Several genes involved in acclimatization protein (Ta.14489.1.S1_at)iron metabolism, starch metabolism, and cell division protease (FtsH, Ta.enzymes (cellulase, catalase, peroxidase, 27445.1.S1_at) were down regulated atlipase, xylanase, peptidase, chitinase),hormone biosynthesis (auxins, gibberellic28 DAA (Figure 6).Figure 6: Differential expression of two major up-regulated genes (peroxidase andC2C2 GATA) and two major down-regulated genes (mutant GBSS1 and PHD)between good (C306) and poor (Sonalika) processing varieties at 3 developmentalstages-7, 14, 28 DAA.11

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE5. The processing quality is affected by size and contrasting wheat varieties, C306 anddistribution of starch granules. Starch Sonalika. These showed more differences ingranules were visualized under light free phenolics than bound phenolics (FigureFigure 7: Starch granules of C306, Sonalika, LOK1 and WH291 at 40X magnification viewed under a lightmicroscope (upper panel) (scale bar = 30µm) and at 1000X magnification under Hitachi VP-SEM S-3400Nfacility (scale bar =50 µm).microscope and scanning electron 8). The wheat variety C306 (86.9 to 104microscope (Figure 7). The four varieties GAE/100g) had more free phenolic contentshowed differences in size distribution. in comparison with Sonalika (63.9 to 68.86. The percentage of amylopectin in the fourGAE/100g).varieties, C306, LOK1, Sonalika and 8. To identify phenolic compunds, freeWH291 was 21.1, 23.8, 22.0 and 25.0, phenolics were analysed on LC-MSrespectively.(Figure 9). The two varieties (C306 and7.Sonalika) showed differences in the types ofFree phenols and bound phenols werephenolic compounds.extracted from the seeds of the twoFigure 8: Free (f) and bound (b) phenolic content intwo Indian wheat varities, C306 and Sonalika, determinedby FCR method (GAE= gallic acid equivalents).Figure 9: Different phenolic compoundsidentified in C306 and Sonalika using LC-MS12

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE1.1.3 Development of Virus Induced GeneSilencing (VIGS)Principal Investigator:Rakesh TuliResearch Fellows:Jitendra KumarJitesh KumarKaushal BhatiIntroduction:1. Detection and characterisation of virusesinfecting wheat in India: The presence ofvirus and insect vector was investigated inVIGS is an approach to facilitate the assessment ofgene function in plants. A recombinant virus thatinfects plant tissue and spreads systemically canbe very useful in the expression of small RNA andsilencing of targeted endogenous genes. Thetarget transcript is degraded by Post-Transcriptional Gene Silencing (PTGS). VIGScan validate the function of a specific gene withina single generation and obviates the need forscreening large populations to identify mutationsin specific genes. Being a transient method, it doesnot require the generation of stable transgenicplants. The project aims at developing an efficientVIGS vector for wheat.wheat growing fields and suspected sampleswere tested (Figure 10). About 80% of thesamples tested positive for the virus in thesamples collected during 2010 and2011(Mohali, Punjab and Wellington, TamilNadu). The whole genome of the virus wassequenced and nucleotide sequences of twoclones were submitted to the GenBankResearch Objectives:under the accession numbers JQ361910 andJQ361911. The virus was named as Wheatl Detection of viruses infecting wheat in Dwarf India Virus (WDIV).India.2. Detection of geminivirus associatedl Analysis of the viral main genome and satellites from infected wheat samples:accessory satellite genomic sequences.The wheat samples found positive for thevirus were investigated, and two types oflllEstablishment of infectivity in differentIndian wheat varieties by inoculating virusin the absesnce or presence of the accessorygenomes.Development of VIGS vector by modifyingthe viral genome.Validation of VIGS vector using visualmarkers.Long Term Objective:Elucidation of functions of candidate genesinvolved in determining the processing andnutritional traits in wheat.Research in Progress:Figure 10: Difference in growth and physicalappearance of healthy and WDIV infected plants.Panelon the right shows probable insect vector.associated satellites (alpha and beta) weredetected (Figure 11).Figure 11: Amplification for a mastrevirus, betasatellite and alpha satellite.13

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE3. Establishment of infectivity in differentIndian wheat varieties: Infectious clonesfor virus and associated satellites were madeand infectivity tests were done. Host rangewas determined by infecting differentIndian wheat varieties which showedconsistent infectivity (Figure 12).Figure 14: Accumulation of virus in thepresence of satellites.V= virus, B= beta satelliteand A= alpha satellite.Figure 12: Agroinoculated wheat plants at 40th day post inoculation. M= mockinoculated and V= virus infectious clone inoculated on different wheat varieties.4. Assessment of effect of alpha and betasatellites on the symptom induction byvirus: The phenotype was observed forassessing the affects of the satellites on thevirus symptom induction process. Theplants were observed showing morewas studied by using microarrays.Expression analysis by using microarrayrevealed that, sixteen unigenes were upregulated by more than 10 fold, thirty fiveunigenes by more than 5 fold and about sixhundred unigenes by more than 2 fold by theFigure 13: Agroinoculated wheat plants at 42nd day post inoculation. M= mockinoculated, V= virus infectious clone and V+A+B= virus+alpha+beta satellite infectiousclone inoculated wheat plants.dwarfism in the presence of satellite incomparison to the virus alone (Figure 13).5. Assessment of effect of alpha and betasatellites on virus accumulation in plants:Semi quantitative PCR was done to assessthe effect of the satellites on the virusaccumulation inside the plant. Theaccumulation of virus was found high in thepresence of the satellites (Figure 14). Theeffects of virus on plant at molecular level inthe presence and absence of these satellitesviral infection. Eighteen different unigeneswere down regulated by more than 10 fold,sixty other by more than 5 fold and sevenhundred other unigenes by 2 fold at theexpression level. Presence of the betasatellite supported the gene regulationpattern of the virus, however, the alphasatellite regulated different sets of unigenes.Presence of both the satellites and virusregulated more number of unigenes in thehost plant (Figure 15).14

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEFigure 15: Gene expression analysis in wheat leaves after 21 dpi with WDIV. (A) Up and down regulation of genes. (B)Venn diagram representing the set of genes up regulated and down regulated during virus infection. (C) Hierarchicalclustering and changes in gene expression in virus only or virus+satellites-infected wheat plants at 21st day post inoculation(dpi). Gene tree maps were generated using k-mean clustering at 2 fold or higher differentially regulated wheat genes.6. Development of VIGS vector by accession number FM998042). The PDSmodifying the viral genome: Two fragment was excised from the cloningmodifications were done in viral genome by vector and was inserted into the MCS ofremoving a small stretch of nucleotides and viral genome using SpeI restrictioninserting Multiple Cloning Sites (MCS) at endonuclease. The viral genome havingthe same positions. The modified viral PDS was termed as VIGS-PDS. In order togenome was cloned in a plasmid vector for infect wheat for silencing of the PDS gene,further manipulation.head-to-tail dimerization of VIGS-PDS wasdone into pCAMBIA1301 binary vector.7. Validation of VIGS vector using visualThe PDS silencing cassette (pCAMBIAmarkers:A 327 bp fragment of phytoeneVIGS-PDS) was transformed intodesaturase (PDS) gene was amplified usingAgrobacterium tumefaciens (straincDNA as a template prepared from wheatGV3101). The empty VIGS vector was also(Triticum aestivum L.). The amplifiedtransformed in Agrobacterium. Thefragment was cloned and sequenced tosilencing cassette and empty VIGS-vectorestablish the identity. Sequence analysiswere inoculated to healthy wheat plantsshowed an identity of 99% with theseparately and scored for phenotype at 15previously reported PDS gene (GenBankdays (Figure 16 A, B and C).TAATATT/AC V E CV2IntronC2C1WDIV2783 bpV1Figure 16: Modification of WDIV for functional tool. (A) Genome organization of WDIV detected fromwheat in India and used for developing as VIGS-vector, C1-Rep Protein, C2-Rep-A Protein, V1-Coat Protein,V2-Movement Protein. (B and C) Leaves from silencing PDS-cassette inoculated (V), empty VIGS-vector(E) and uninoculated control (C) wheat plants. (C) Partial view of a VIGS-PDS inoculated wheat plant.15

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEResearch in Progress:1. Establishment of in-vitro regeneration:a. Callus mediated regeneration:1.1.4 Efficient genetic transformation of wheatPrincipal Investigator:Siddharth TiwariResearch Fellow:Anshu AlokIntroduction:Efficient regeneration and genetic transformationprotocols are pre-requisite for crop improvementthrough genetic engineering. The protocols forcallus as well as direct multiple shoot mediated in-i. In the four high yielding (PBW621,PBW550, HD2697 and LOK1) cultivatedvarieties of wheat, matured embryoscultured on MS medium supplementedwith 2, 4-D showed the best response forcallus induction (Figure 17 A and B).ii. Multiple shoot induction and elongationfrom callus were observed to be mostefficient on MS medium supplementedwith zeatin (Figure 17 C, D and E).Figure 17: Callus induction and regeneration. (A) Explant (mature embryo).(B) Callus formation. (C-E) Stages of regenerated calli. (F) Root formation.vitro regeneration and Agrobacterium-mediatediii. Healthy roots were induced on the MSbasal medium (Figure 17 F).genetic transformation of wheat have beenoptimized. Transgenic plants with reporter b. Direct multiple shoot mediatedb-glucuronidase (GUS-Intron) gene expression regeneration:have been developed and further molecular and i. Mature embryos of high yieldingsegregation studies are in progress. The overallaim of the work undertaken is to establish anefficient in-vitro regeneration and genetictransformation protocols for wheat.Research Objective:Establishment of efficient in-vitro regenerationand genetic transformation protocols for wheat.Long Term Objective:Development of nutritionally rich andagronomically improved varieties.(PBW621, PBW550) cultivars of wheatshowed best direct multiple shootFigure 18: Direct multiple shoot regeneration of wheatplants. (A) Multiple shoots. (B) Root formation.16

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEinduction without an intermediary calluso n M S m e d i a c o n t a i n i n g 6 -B e n z y l a m i n o p u r i n e ( B A P ) a n dThioridazuron (TDZ) (Figure 18 A).ii. Culture was maintained as the continuoussource of shoots.iii. Shoots were separated from the multipleexpression studies in transgenic wheat.a. Callus based regeneration oftransgenic wheat: The transient andstable expression of reporter gene wasnoticed as blue colour on the embryosand callus-derived shoot bud,respectively (Figure 19 A and C). NoFigure 19: GUS histochemical assay. (A) Transientexpression in embryos (blue color). (B) Non-transgenicembryos. (C) Stable expression in callus-derived shootbud (blue color). (D) Non-transgenic shoot bud.shoot clumps and rooted on MS basalmedium containing Indole-3-butynic acid(IBA) and Naphthaleneacetic acid (NAA)(0.5mg/l each) (Figure 18 B).2. Establishment of genetic transformation:Several factors were considered for theoptimization of Agrobacterium-mediatedgenetic transformation. Mature embryoderived callus and direct multiple shootinduction based in-vitro regeneration wasused for reporter (GUS-Intron) geneexpression of GUS was noticed in thecontrol embryos and shoot bud (Figure19 B and D).l· Direct multiple shoot induction basedregeneration of transgenic:a. Stable expression was noticed as bluecolor on the shoots developed withoutan intermediate callus (Figure 20 A andB). No expression of GUS was noticedin control shoots (Figure 20 C).Figure 20: GUS histochemical assay. (A and B) Transformed germinated shoots (blue color). (C) Nontransformedshoots.17

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE1.1.5 Metabolic engineering of phytic acidpathway to enhance iron bioavailability in wheatPrincipal Investigators:Ajay K. PandeyCo-Investigator:Siddharth TiwariResearch Fellow:Research Objectives:lllIdentification of genes involved in phyticacid pathway.Comparative gene expression analysisduring different developmental stages of thewheat grains.Construction of RNAi vectors to inhibit theexpression of the target genes in wheatplant.Roohi BansalIntroduction:l Stable transformation and expression (forRNAi) in wheat plants using selectionmarkers.Phytic acid (myo-inositol-1, 2, 3, 4, 5, 6-l Examination of quantitative relationshiphexakisphosphate; PA; InsP6) beside been a between total phytate content and inorganicstorage form of phosphorus in seed is also a well phosphorus in silenced wheat plants.Figure 21: Schematic representation of PA pathway in plants.Long Term Objectives:Developing wheat varieties with reduced totalphytate content and enhanced bioavailable iron.Research in Progress:1. We analysed transcriptome data for thegenes which are differentially regulated andpossibly involved in phytic acid synthesispathway. During our initial analysis of C306and LOK1 varieties of wheat grains, wewere able to identify some genes whichputatively codes for inositol phosphatekinases (Table 1).know seed anti-nutrient. The chelating properties 2. Blastx analysis of the corresponding geneof PA in seeds contribute to reduced iron and other ID suggests that they belong to inositolmicronutrients bioavailability. One of the major phosphate kinase (IPK) gene family. All thechallenges faced by researchers worldwide is how genes mentioned below belong to the lateto increase the bioavailability of iron in wheat phase of the InsP synthesis pathway.seeds. In this project we want to utilize functional 3. Wheat being a hexaploids crop (A, B and Dgenomic tool/s to address the role of phytic acid genome), we are designing primerssynthesis genes. Our goal is to reduce the PA targeting at the conserve nucleotide regioncontent by silencing late PA pathway genes to amplifying different genes by utilizing(Figure 21) in wheat grains and therefore genomic DNA as a template. Further, RNAianticipating an increasing bioavailability of total constructs will be developed to target theseiron.genes to assess their functional role in PAsynthesis18

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTETable 1: List of putative phytic acid biosynthesis pathway genes regulatedduring the anthesis in wheat plants.Probe set Gene Bank ID Annotated function(based on blast homology)Ta.27561.1.S1_at CK215630 inositol-tetrakisphosphate 1 -kinase 1 (from Zeamays)Ta.30464.1.A1_at CK214494 inositol hexaphosphate kinase (from Arabidopsis)Ta.5574.1.S1_a_at BJ227701 1-phosphatidylinositol-3-phosphate 5 -kinase-like(from Oryza sativa Japonica)TaAffx.113052.1 .S1_at CA618510 inositol-tetrakisphosphate 1 -kinase 1 (from Zeamays)TaAffx.121326.1.S1_at CA690518 inositol 1,3,4 -trisphosphate 5/6 -kinase (fromOryza sativa Japonica)TaAffx.19998.1.A1_at CD867474 inositol phosphate kinaseTaAffx.112860.1.S1_at AF532601 multidrug resistance associated like protein :MRP2 (Zea mays: lpa-1 gene)1.2 Accelerated breeding for qualityimprovementPrincipal Investigator:Monika GargResearch Fellows:Rohit KumarVishal GuptaShaweta AroraIntroduction:is well known. The protein content and typesdetermine the end product quality like bread,biscuit, cake, chapatti and noodles etc. Biscuitmaking requires soft wheat with low proteincontent and specific combination of differentalleles (2+12 allele of High Molecular Weightglutenin subunit gene (HMW-GS) at chromosome1D (locus GluD1), Pina-D1a, Pinb-D1a alleles ofpuroindoline gene etc). Bread making requireshard wheat with high protein content and specificcombination of different alleles (5+10 allele ofGluD1-HMWGS, Pina-D1b, Pinb-D1a/b etc).Chapatti making requires medium strength wheatwith medium protein content. The contribution ofdifferent genes/alleles to chapatti making ispoorly understood.In the developed countries, grain market is drivenby wheat quality. A wheat class/grade is awardedto a product based on its processing and end-usequality. Validated markers are available for eachproduct type and are being routinely used. Indiancultivars are released based upon agro climatic Research Objectives:zones, time of sowing and soil fertility. Validatedmarkers are not available for a major product i.e.lchapatti. Available validated markers are notbeing utilized. In India, there is a need of breedingcultivars, based upon their processing qualityl Marker discovery.(milling and baking characteristics), markerdevelopment and utilization of validated markers.lGeneration of breeding material withimproved processing quality.Generation of near isogenic lines for markerdiscovery.Processing quality of wheat depends upon seeds l Generation of RIL populations to validateharvested from field and its components like markers.proteins, starch, non starch carbohydrates andlipids. Protein's contribution to processing quality19

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTELong Term Objectives:lines. PCR multiplexing for selection ofgrain softness genes in the backcross linesl Development and validation of markerswas standardised.associated with different processing qualitytraits. 2. Accelerated breedingl Screening of generated breeding material For improvement of bread making qualityfor multiple traits and introduction into we are utilizing wild species of wheat andmultinational trials with the help of their genetic stocks (addition lines,Directorate of Wheat Reaearch (DWR)/ substitution lines and translocation lines) asState University scientists to design end donors. For improvement of biscuit makingproduct (bread, biscuit, chapatti) specific quality, we are utilizing two soft wheatcultivars.landraces NAP HAL and IITR67 as donorsResearch in Progress:of grain softness. For chapatti making oldcultivars C306 and LOK1 (well known for1. Marker discoverytheir good chapatti quality) are beingutilized. Factors/genes from the donors areThree traits (bread, biscuit and chapattibeing transferred to agronomically superiormaking) and the factors responsible for theirbackground by accelerated breedinggood quality are under study. Transapproach.criptomics studies at different developmentalstages of good (C306 and LOK1) a. For improvement of chapatti makingand poor chapatti making varietiesquality, good chapatti making old(Sonalika and WH291) indicated that cultivars were crossed with high yieldingamong the earlier reported genes affectingrecent cultivars (PBW343, PBW550 andwheat processing quality, granule boundPBW621). Crossed seeds werestarch synthase 1(GBSS1) was several foldsbackcrossed at DWR Regional Station,down-regulated in good chapatti varieties.Dalang Maidan, Lahaul, HimachalGBSS1 is involved in amylose starchPradesh during off season. One hundredsynthesis. Genomic variation of GBSS1 wasand eighty nine BC 1 plants were screenedstudied in several wheat cultivars. Allelicfor absence of GBSS-1B and out of themvariation of GBSS1 in Indian cultivarssixty plants were selected. Negative plantsindicated that GBSS-A1 and GBSS-D1 geneswere backcrossed with selected cultivarswere non-polymorphic and present in all theand BC 2 plants have been sown at Dalangcultivars studied. GBSS-B1 gene wasMaidan.polymorphic based on presence/absencealleles. Preliminary study on different b. For improvement of biscuit makingcultivars and lines with known chapattiquality, donor landraces were againmaking quality indicated that absence ofcrossed with high yielding recent cultivarsGBSS-B1 gene was co-related with good(PBW343, PBW550 and PBW621).chapatti making quality (Figure 22). AmongCrossed seeds were backcrossed atstarch pasting characteristics, breakdown inDalang Maidan during off season. Oneviscosity was found to be co-related withhundred and seventy eight BC 1 plantsgood chapatti making quality. Validation ofwere selected based upon presence ofrelationship between GBSS1 and (Rheopuroindoline gene PinaD1a and fiftyVisco Analyzer) RVA-breakdown ofselected plants were backcrossed withviscosity in chapatti making is in progress. selected cultivar. BC 2 plants have beenFor improvement of biscuit making qualitysown at Dalang Maidan.major genes (puroindoline and HMW c. For improvement of bread making quality,glutenin genes) responsible for grainwild species/genetic stocks of Ag.softness were characterised in selectedelongatum, Ae. searsii and Ag.20

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEintermedium, are being utilized. We wantto transfer HMW-GS genes related to highgrain strength from wild species tochromosome 1A of wheat (translocationlines) as later has some alleles thatcontribute negatively to bread makingquality. For this purpose marker assistedselection and transfer to agronomicallysuperior cultivars is in progress. Theendosperm half of seeds of the abovegenotypes were screened for storageproteins of interest by sodium dodecyls u l p h a t e - p o l y a c r y l a m i d e g e lelectrophoresis (SDS-PAGE). Theremaining half seeds expressing theselected wild species HMW-GSs wereraised into plants and crossed with theselected Indian wheat varieties. The aboveprocedure was repeated one more time atNABI and the selected seeds were grownat Dalang Maidan. The seeds werereselected and sown at NABI fieldFigure 22: Marker assisted breeding for processingquality. (A) SDS-PAGE of HMW-GSs of endospermhalf of seeds to find individual plants carrying additionsubunit from wild species (indicated by arrow) havingpositive effect on bread making quality. (B) PCRmultiplexing for biscuit making quality to co-amplifypuroindoline gene and high molecular weight gluteningenes. (C) PCR multiplexing for chapatti makingquality to co-amplify GBSS-1B functional and nonfunctionalalleles.1.2.1 Development of puroindoline genedatabase of Indian cultivarsPuroindoline genes have been reported to bestrongly associated with grain softness; a traitassociated with processing quality of wheat. Twogenes Pina-D1 and Pinb-D1 located onchromosome 5D have been found to be associatedwith grain softness. Pina-D1 and Pinb-D1 encodesmall (13 kDa) protein called puroindoline A andB (PINA and PINB), which exhibit thecharacteristics of alpha helical structure, tencysteine residue, a tryptophan rich domain andbasic nature (PI 10.5). The soft texture of thecommon wheat requires both wild type PINA andPINB. Diverse alleles with single nucleotidepolymorphism (SNPs), small insertions/deletionsand/or megabase deletions have been associatedwith hard texture of common wheat. There is noinformation available on sequences ofpuroindoline genes in the Indian germplasm. Thisstudy was planned to generate a database ofpuroindoline genes of around 500 Indiancultivar/lines that can be utilized by Indianbreeders. This will also help to find out structurefunction relationship and involvement of differentgenes in grain hardness.Research in Progress:1. PCR amplification of Pina-D1gene fromfive hundred and fifty one Indiangermplasm indicated amplification and thuspresence Pina-D1a functional allele in fiftythree lines i.e. 9.6% of total lines.2. Sequencing of some of the expressed Pina-D1a allele indicated no sequence variationin the Indian germplasm. PCR amplificationof Pinb-D1gene from five hundred and fiftyone Indian germplasm indicatedamplification in all the lines studied (Figure22).3. Sequencing of Pinb-D1 gene from differentIndian cultivars indicated sequencevariations. Out of thirty three linesanalysed, twenty six lines had Pina D1b(null) allele and functional Pinb-D1a allele.These varieties included RAJ4120, HS505,VL616, K0307, WH1062, HD3002,21

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTETable 2: Indian cultivars having mutated Pin b-D1gene with mismatch position andchanged nucleotide and /or amino acid.S. No. Cultivar Allele No. ofentriesMismatch positionNucleotide /Amino acidBasechange/AA change1 VL934 Pin b-D1b 1 223/ 46 G-----A/ G-----SAllelePublished2 DBW17 Pin b-D1y1 1 92 T-----C New3 WH1073 Pin b-D1y2 1 93/2 T-----A/V----A4 HS511 Pin b -D1y31 256/57 T-----C /C----R5 HP1102 Pin b-D1y4 1 52 T----A New6 PBW613K0710NewNewPin b-D1y5 2 53 T----A NewMACS6222, NP809, HS1138, K816, substitution without change in amino acid.KSML3, NARMADA195, UP215, Allele Pin b -D1y3 and Pin b -D1y4 resultedHUW12, HW5210, NP836VL916, VL921, in amino acid substitutions at positions 2ndHS508, VL829, HPW251, UP2771, HS295, and 57th. These mutations were not withinHS512, HS513, HPW296, VL924, VL925, tryptophan rich domain (34th to 47th aminoVL935, UP2772, HD2967, PBW343, acid) that leads to functional change of thePBW621, WH1061, PBW628, WH1081, Pin b-D1 gene. 78% of Indian cultivars haveHD3014, UAS315, K0607, DBW39, wild type Pinb-D1a allele.HUW612 and DBW46.6. Mutant Pinb-D1b allele commonly found in4. One line VL934 had Pina-D1a (functional Australian wheat is rare in Indian cultivars.allele) and Pinb-D1b (non-functional Five new alleles have been identified inallele). Pinb-D1b allele resulted from Indian germplasm that may contribute moreGuanine to Adenine nucleotide mutation to the sources of variation. We haveleading to glycine to serine amino acid sequenced only thirty three cultivars formutation at 46th position as reported in Pinb-D1 gene. Currently work is in progressliterature.to sequence Pina-D1 and Pinb-D1gene.Others genes in the pipeline are GSP and5. In addition five new mutations (Pin b-D1y1Pinb-2D1 that are known to be involved into Pinb-D1y5) were identified as listed ingrain hardness.Table 2. Allele Pin b-D1y1, Pin b-D1y4 andPin b-D1y5 showed single nucleotide22

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE2. IMPROVING FRUITS FOR POST HARVESTQUALITY AND NUTRITION2.1 Genetic transformation of banana for Research Objectives:quality improvementl Development of efficient geneticPrincipal Investigator:transformation in Indian banana varietiesSiddharth Tiwariviz., Grand Nain and Rasthali.lCo- Investigator:Evaluation of fruit specific promoters andpro-vitamin A candidate genes andRakesh Tulicombinations there of after transformationResearch Fellow:in Indian banana.Anshu Alokl Development of transgenic lines usingIntroduction:constructs for pro-vitamin A biofortification.l Initial field and nutritional analysis of fruitBanana and plantain are among the most under containment conditions.important fruit crops in the world and contributelsignificantly towards the food security in many Selection of lines for nutritional anddeveloping countries. India is the largest producerbioavailability analysis.of banana in the world with an annual production Long Term Objectives:of about 18 million tonnes. Banana are consumedDevelopment of pro-vitamin A rich biofortifiedfresh (except cooking types (plantain)) withoutIndian bananas, bioavailability study, nutritionalany thermal cooking and processing. The pulp of analysis and agronomical filed trials ofdessert banana remains unexposed to light, hence transgenics.allowing minimal degradation of carotenoids inthe edible flesh. Genetic improvement of banana Research in Progress:through conventional methods is limited because 1. Germplasm collection and plantation atmost of the cultivated varieties are triploid in NABI research field:nature, hence sterile and provide natural barrier toa. Suckers of established banana cultivarscross pollination. Therefore, transgenic(Grand Nain, Robusta, Nendran, Poovan,approaches hold high promise for developingRasthali, Red Banana, Ney Poovan,sustainable resistance against diseases/pests and Virupakashi, Karpuravalli, Dwarffor biofortification in banana. Plant regeneration Cavendish, Dwarf-Robusta, Udhayam andof banana has been reported from various explant Nanjanagud Rasabale) have been collectedsources. Immature male flowers are the most from Tamil Nadu Agricultural Universityresponsive material for initiating the embryogenic (TNAU), Coimbatore and Gandhi Krishicultures. An Embryogenic Cell Suspension (ECS)Vigyana Kendra (GKVK) Bangalore(Figure 1A). The collected suckers haveculture is being used for Agrobacteriumbeen grown at NABI research field fortumefaciens mediated genetic transformation inestablishing germplasm (Figure 1B and C).many laboratories. However, genetictransformation and regeneration frequencies are 2. Establishment of ECS culture for geneticreported to be highly genotype dependent. Thus,transformation of banana:optimization of transformation protocol for any a. Immature male flower buds of two cultivarsparticular type of cultivar becomes a prerequisite (Grand Nain and Rasthali) were collectedfor genetic improvement in that cultivar.from TNAU, Coimbatore and NRCB,23

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTETrichy, Tamil Nadu (Figure 2 A and B). d. Best response for callus formation wasb. Immature male flower hands of rank 1 to 15optimized (Figure 2 D).adjacent to the floral apex were isolated and e. Optimization of ECS has been initiated forcultured on MS medium containing several the induction of globular embryos andFigure 1: Establishment of Banana germplasm at NABI research field. (A) Bananasuckers. (B) Banana germplasm. (C) Banana plant with fruit at NABI research field.combinations and concentrations of genetic transformation (Figure 2 E).different growth regulators for thef. ECS was used for transient transformationoptimization of protocol (Figure 2 C).of reporter (GUS-Intron) gene andc. Calli were formed on the callus forming expression was noticed as blue colour on themedium. However, efficiency and response ECS (Figure 3 A and B). No expression offor callus induction depends upon the GUS was noticed in control ECS (Figurecultivars.3C).Figure 2: Stages of ECS and culture development for somatic embryogenesis. (A, B) Immature maleflower bud. (C) Immature male flower hands of rank 1 to 15 adjacent to the floral apex. (D) Embryogeniccallus induction. (E) Suspension culture.Figure 3: Transient genetic transformation of ECS. (A) ECS (B) Transformed suspensionculture cells (blue clour). (C) Non-transformed suspension culture cells (no blue clour).24

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEFigure 4: Different stages of micropropagation of banana. (A) Banana sucker as explants. (B) Sucker turnedinto green bud. (C) Induction of multiple shoots. (D) Shoot elongation. (E) Tissue culture raised plants.3. Establishment of micropropagation: for fresh fruits. The concept of globala. Suckers of Grand Nain, Rasthali and Dwarfharmonization of quality standards is emergingCavendish cultivars were used forrapidly which warrants the development ofoptimization of protocol for microglobally acceptable analytical tools for qualitypropagation (Figure 4 A).assessment and delivering the logical andsustainable solutions to maintain postharvestb. Multiple shoots were induced and quality in the supply chain. The genetic,multiplied under some of the culture physiological and biochemical diversity in theconditions (Figure 4 B, C and D).fresh produce require the commodity specificapproach to develop postharvest solutions.c. Tissue culture raised acclimatized plantshave been generated and transferred atNABI research field (Figure 4 E).2.2 Quality enhancement and postharveststability of tropical fruitsResearch Objectives:lUnderstanding and explicating thecomplexity of biochemical and molecularfactors contributing to produce quality.Principal Investigator:l Development of preharvest and postharvestSukhvinder P. Singhstrategies to improve quality and enhanceResearch Fellow:postharvest stability of farm produce.Manpreet Kaurl Translation of knowledge and innovationIntroduction:into commercially important products/processes/services for primary producersQualitative and quantitative postharvest losses of and industry.fresh fruits are significant in India. Enhancing andmaintaining quality through adaptive preharvest Long Term Objectives:and postharvest actions is important to reducel To develop protocols to address commoditylosses, meet consumer expectations and promote specific quality and postharvest issues.domestic and international trade. The primelobjective of this research program is to understandTo improve and maintain farm producebiological basis of fruit quality and generate basicquality through application of Generallyknowledge to develop pre and postharvestRecognised As Safe (GRAS) compounds,strategies for extending the storage stability andnatural metabolites, growth regulators andmaintaining produce quality. The research effortsantagonistic microorganisms.are directed to expand the horizon of l Generating primary data on the producemeasurement, definition and meaning of 'quality' quality with special reference to flavour and25

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEnutritional quality.Research in Progress: Mango, citrus, litchi andguava are the targeted crops to study the qualityand postharvest stability of fruits.Mangolcyclopropene (1-MCP), which is anethylene action inhibitor, was not effectiveto retard the process of fruit softening inboth treated and control fruit. Theinefficacy of 1-MCP is contrary to its proveneffectiveness in other fruits.Mango export from India has been jeopardised by 4. The qualitative profiling of aroma volatilesthe postharvest phytosanitary requirements by in mango showed that sesquiterpenes andseveral countries such as the United States of monoterpenes constituted about 85-90% ofAmerica, Japan, Australia and New Zealand. In the total volatiles production. With therecent years, heat-based phytosanitary treatments progress of fruit ripening, the abundance ofhave received regulatory approvals from Japan, sesquiterpenes decreased and that ofAustralia and New Zealand which led to the monoterpenes and esters increased.establishment of these facilities in the major5. The evolution of aroma volatiles such asmango producing and exporting zones of themonoterpenes and esters was faster in VHTcountry. However, there are certain qualityfruit, parallel to other changes in fruitconcerns associated with these treatments whichripening, compared to control.can affect consumer acceptance of the exportedfruit in the international markets. Our objective 6. The abundance of esters (fruity odour)was to assess the impact of heat treatment declined significantly in VHT fruit after 8protocols on quality and postharvest behaviour of days of shelf-life at 26°C. The VHT wasmango fruit.effective to reduce the microbial on fruit1.surface and also helped reducing theIn 2011 season, an experiment on Vapourincidence of anthracnose disease.Heat Treatment (VHT) of 'Chausa' mangowas conducted in a commercial VHT 7. The VHT fruit stored at 10°C for 2 or 3facility at Saharanpur, U.P.weeks and then held for 4 days at 26°C forsimulated shelf conditions showed lower2. The results showed that the VHT acceleratedincidence of chilling injury. However, therate of fruit ripening, resulted in developdetrimentaleffect of VHT in terms of fasterment of uniform skin colouration (Figure 5).fruit softening was not prevalent in fruit heldThe heat treated fruits reached ready-to-eatin cold storage after treatment.Figure 5: Effect of VHT on skin colouration in'Chausa' mangoes after 48 hours at 26°C.stage within 4 days of treatment against the 8days in control held at ambient conditions(26°C).3. Post-VHT treatment with 1-methy-In future, the efforts will be made to minimise thedetrimental effects of VHT on mango fruit qualityattributes and also to retard the process of fruitripening through better understanding of fruitphysiology.LitchiSkin colouration in litchi is due to the presence ofanthocyanin pigments. whose biosynthesis andpostharvest stability can seriously affect fruitquality. Targeted metabolite profiling of litchi(cvs. 'Calcuttia', 'Dehradun' and 'Seedless') wasconducted at different stages of maturation (green,colour break/pink, and red).1. The results showed that the concentrations26

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEFigure 6: Chromatograms representing the anthocyanins (top) in the pericarp andsugars (bottom) in the aril of litchi fruit.of chlorophyll a, chlorophyll b, xanthophyll tissue showed that the concentrations ofand pheophytin in the pericarp tissue fructose, glucose and sucrose increaseddeclined during maturation in three significantly during maturation. Fructosecultivars.was found to be the principal sugar followedby glucose and sucrose in all cultivars.2. Figure 6 represents the chromatograms of'Seedless' cultivar showed the highestvarious targeted metabolites such asconcentration of sucrose at the red-ripe stageanthocyanins and sugars. Cyanidin-3-ocomparedto other cultivars, 'Dehradun' andrutinoside (Cyn-3-o-rut) was identified and'Calcuttia'. Among organic acids, malic acidquantified as the major anthocyaninwas the predominant acid in all threepigment in all three cultivars and wascultivars. The other organic acids includeddetected during colour break/pink stage andcitric acid, succinic acid, tartaric acid andshowed increasing trend during maturation.shikimic acid. The concentration of malicThe concentration of Cyn-3-o-rut wasacid and other minor organic acids declinedmaximum in 'Dehradun' cultivar followedwith the progress of maturation of aril tissue.by 'Calcuttia' and 'Seedless'.The protocols developed for identification3. The other anthocyanin pigments were and quantification of these metabolites willidentified as cyanidin-3-o-glucoside, be used for objective assessment of thecyanidin-3, 5-diglucoside and cyanidin-3-o- effects of various postharvest treatments ongalactoside. The sugar profiling of fruit aril litchi fruits27

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE3. BASIC BIOLOGY FOR CROP IMPROVEMENT3.1 Biology of seed development in custardapple and litchiPrincipal Investigator:Rakesh TuliCo-Investigator:Sudhir P. SinghResearch Fellow:Yogesh GuptaIntroduction:28carpels (>100). Each carpel has a singleanatropous ovule that may develop into a singleseed. The Annona fruit develops from the clusterof fertilized carpels, thus the aggregate fruitcontains several fruitlets. Out of the multiplefruitlets a few fruitlets develop naturally, withoutseeds. Annona squamosa produces fruits withgreater number of seed fruitlets, sixty-eightyseeds per fruit. A contrasting genotype, namedNMK-1, has been identified having fewernumbers of seeded fruitlets and many of thefruitlets are seedless (Figure 1). The varietyselected from the seedling population of Annonasquamosa by a farmer, Shri N. M. Kaspate ofMadhuban nursery, Solapur, Maharastra.Seeds in many fruit crops like custard apple, litchi,guava, orange, mango and grape are a hindrance tofruit processing and fresh fruit consumption.Sugar (or Custard) apple, a popular fruitThe aim of the project is to understand thethroughout the tropics, belongs to genus Annona.molecular basis of the development of seeded andThe genus Annona contains about one hundredseedless fruitlets in the same fruit of Annonaand twenty species, out of which six speciessquamosa. The major research objective is to doproduce edible fruits; Annona squamosa (Sugartissue specific differential transcriptomics inapple or Sarifa or Sitafal), Annona reticulatadeveloping fruitlets of Annona species for(Custard apple or Bullock's heart or Ramphal),identifying genes involved in seed development.Annona cherimola (Cherimoya or Hanumanfal),Annona muricata (Sour sop), Annona atemoya Litchi is another crop where seedlessness is a(Laxmanphal, a hybrid between Annona desirable trait. The inflorescence is composed ofcherimola and Annona squamosa), Annona several panicles bearing three types of flowersdiversifolia (Ilama) (Gandhi and Gopalkrishna, which open in succession with the same panicle.1957). Annona squamosa is the most popular The three types of flowers are (type 1) male flowerspecies in India. The Annona species flowers which has functional anthers but lacks pistil, (typecomprise of a cluster of stamens (>200) and2) hermaphrodite female flower which hasACcompletely developed pistil but the anthers donot dehisce and contain very less number ofviable pollen. The ovary is bicarpellary, eachcontaining an ovule, (type 3) hermaphroditeBDmale flower is a functional male, which haspistil but lacks stigma and thus pollen fail toenter. The litchi fruit develops from type 2flowers. Generally only one carpel of type 2flower gets developed into fruit afterFigure 1: Seed percentage in the contrasting cutivars of pollination. Botanically the fruit is a drupe.custard apple. (A) Flower of Annona squamosa withmultiple stamens and pistils. (B) Seed percentage in the The edible part of the fruit is called as aril (orfruits of Annona squamosa and NMK-1. (C) Fruits of pulp) which is an outgrowth of the outercontrasting genotypes of Annona squamosa and (D)NMK-1.integument. Some litchi accessions, popularly

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEknown as 'Seedless' or 'Bedana', have seeds ofvery small size with well-developed fleshypulp, in comparison to the common litchivarieties (Figure 2). The project aims toidentify genes related to small size of seeds bydoing tissue specific differential transcriptomicsof contrasting accessions of litchi.Research Objectives:lLong Term Objectives:llTo identify genes responsible for thedevelopment of fruits and seeds.To develop different varieties of seedlessfruit crops.Research in Progress:Differential transcriptomics of develop-ing fruits of contrasting genotypes ofAnnona species.1. A genotype, NMK-1, of Annona hasbeen identified with more number ofseedless fruitlets at Madhuban Nursery,Solapur, Maharashtra.2. Self-pollination and collection ofl Tissue specific differential transfertilizedcarpels (at different days aftercriptomics in ovules of contrastinggenotypes of Annona species at differentpollination) of contrasting genotypes,developmental stages.NMK-1 and common custard apple(Annona squamosa) is in progress atl Tissue specific differential trans- Solapur, Maharashtra.criptomics in ovules of contrasting litchi3. Litchi contrasting genotypes, showingaccessions, at different developmentaldifferences in seed development andstages.seed size are being screened forl Accessions of litchi, showing candidate genes to develop moleculardifferences in seed development and approaches to seedlessness. Polymorph-seed size to be screened for candidate ism in nucleotide sequences of agenes to develop molecular approaches candidate gene for seed developmentto seedlessness.has been identified (Figure 2). Furtherstudy on molecular details is in progress.Figure 2: Polymorphism in nucleotide sequence of a gene related to seedlessness indiverse accessions of litchi29

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE3.1.1 Development of approaches for the technology to prevent the development of seeds inmodulation of seedlessness through rootstock fruit crops, where seed may not be desirable bysignalingfood processing industry. Prevention of seedPrincipal Investigator:development may also lead to enhancing theproductivity and biomass. The objective of theRakesh Tulipresent proposal is to examine if siRNA basedCo-Investigator:Sudhir P. Singhsignaling can be applied to develop rootstocks,which may influence seed development in thescion, grafted on to the stock. The rootstock wouldResearch Fellow:transmit siRNAs, targeted for silencing of specificAnita Kumarigenes involved in the development of embryo andIntroduction:endosperm. Effect of silencing on thedevelopment of fruits and seeds on the graftedFruit crops are mostly propagated through asexualscion will be studied, using the model plantreproduction by grafting. In grafting, one plant isArabidopsis thaliana (Fugure 3). If successful, theselected for its roots and is called rootstock. Thestudy will be applied to a suitable horticulturalother plant is selected for its stems, leaves, flowerscrop plant.Research Objectives:llTo develop various transgenic plantsharbouring RNAi constructs targeting thegene expression in ovule involved in seeddevelopment.To examine the movement of RNAidelivered from root-stock to scion affectingexpression of genes in Arabidopsis.Long Term Objective:l Development of improved rootstock for theFigure 3: Stem grafting in Arabidopsis thaliana.delivery of long distance signals into thescion.or fruits and is called the scion. Scion–rootstockinteraction affects the physiology and thusResearch in Progress:phenotype of the scion plant. For example, to 1. Transgenic Arabidopsis lines expressinginduce dwarfing in apple trees, scions are grafted reporter gene (GUS) have been developed.on to dwarf root stock to give smaller canopy, so Breeding of transgenic plants to obtainthat it becomes suitable for planting the trees at homozygous lines is in progress.higher density. In recent years, evidence2. Development of transgenic Arabidopsissuggestive of long-distance transport of signalslines expressing reporter gene in ovule is inthrough vascular system have been reportedprogress.appears to be increasing. Such signals caninfluence various developmental and 3. Transgenic Arabidopsis lines expressingphysiological processes such as flowering, (constitutive and phloem specific) doubletuberization, nodulation, leaf development, shoot stranded RNA- hairpin homologue ofbranching and disease resistance.reporter gene have been raised. Breeding oftransgenic plants to obtain homozygousThe research project anticipates establishment of a lines is in progress.technology which can lead to the development of a4. Transgenic Arabidopsis lines expressingnew approach to use transgenic rootstocks for(constitutive and phloem specific) doubledelivering specific siRNAs in non-transgenicstranded RNA- hairpin homologue ofscions for the modification of economicallycandidate gene of seed development haveimportant traits. In the present case, the principlebeen raised. Breeding of transgenic plants towill possibly lead to the application of theobtain homozygous lines is in progress30

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE4. DIET AND HEALTH4.1Effect of dietary constituents onadipogenesis4.1.1Role of arabinoxylans from millets inregulating adipogenesis: A nutrigenomic studyPrincipal Investigator:Kanthi K. KiranCo-Investigator:Mahendra BishnoiIntroduction:Current anti-obesity medications arepharmacological agents which can reduce orcontrol weight. These drugs affect one of thefundamental processes of the weight regulation inhuman body i.e. altering appetite, metabolism orconsumption of calories. There is only one anti-obesity medication, Orlistat, approved by FDA. Itacts through inhibition of pancreatic lipaseenzyme. Rimonabant, acting through blockade ofthe endocannabinoid receptor and Sibutramine,a c t i n g t h r o u g h b r a i n b y i n h i b i t i n gneurotransmitter metabolism were previouslyapproved drugs which have been withdrawn fromthe market in several countries and regionsObesity is a worldwide epidemic and has asignificant negative impact on health, mortalityand related costs. Diet and genetic factorsincluding gene mutations are closely related to thedevelopment of obesity. Untreated, it may lead tochronic diseases, such as diabetes (type II),hypertension, cardiovascular disease and certaintype of cancers. Present anti-obesity medicationsare shown to be associated with side effects uponlong term usage. Phytochemicals such aspolyphenols and non-starch polysaccharides havetremendous health benefits. Arabinoxylans (AXs)are non-starch hemicellulosic polysaccharides,major constituents in the cell walls of cereals andmillets, are reported to have potential healthbenefits in alleviating disease symptoms such asdiabetes, atherosclerosis and colon cancer. WheatAXs help in regulating the weight gain and obesityby controlling the release of gut hormones likepeptide YY and Glucagon-Like Peptide-1(GLP1). The potential beneficial effects of AXsfrom finger millet and kodo millet in regulatingadipogenesis and their metabolic fate in animalmodels will be studied, which is expected toprovide the insight on mechanism of action ofAXs in adipogenesis regulation and help indesigning functional foods with enriched Axs.Research Objectives:llMechanistic studies on the role of AXs onadipogenesis using mouse 3T3-L1 cell linesin-vitro.Modulating adipogenesis and obesity usingAXs in high fat diet induced obese mousemodel through molecular approaches.Long Term Objectives:llTo study the role of high dietary fiberconsumption and the role of gut microbiota inalleviating weight gain and obesity.Designing AXs enriched functional foodmodels that can modulate metabolicdisorders.Research in Progress:Currently, we are standardizing the culturing ofpreadipocytes and differentiation into adipocytes.Primers for important transcriptional factors,enzymes in the fatty acid pathway and keyadipokines are designed. Chemo-enzymaticextraction of AXs from finger millet grains isunder progress.4.1.2 Transient Receptor Potential (TRP)channel mediated modulation of adipogenesisand obesity by dietary moleculesPrincipal Investigator:Mahendra BishnoiCo-Investigator:Kanthi K. KiranIntroduction:31

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEincluding India (Banned Medicines, press l To study the effect of dietary modulations ofrelease), Ministry of Health and Family Welfare. TRP channels (TRPV1: capsaicin, capsiate,thFebruary 10 , 2011). The potential side effects piperine; TRPA1: garlic, cinnamon; TRPM8:(increased cardiovascular concerns, stroke, menthol) in a diet (high fat) based in vivosuicidality and depression) of these drugs are mouse model of obesity.much more than their beneficial effects. Over theyears it has been seen that the best and mostLong Term Objectives:effective options available for overweight and l Developing diets constituted of modulatingobese individuals are dieting and physical food components and study their effect onexercise. It is important to have dietary regulations adipogenesis, obesity and relatedto prevent life style problems rather than to search complications.for the treatment. Available literature suggests thatl Special dietary formulations will be madeTransient Receptor Potential (Transient Receptorusing TRP channel modulating dietary agentsPotential Vanilloid (TRPV1), Transient Receptoralone or in combination with other nutrients.Potential Ankyrin (TRPA1), Transient ReceptorPotential Metastatin (TRPM8) channels are l Undertake clinical trial using humanpossible candidates to regulate energy metabolism volunteers for establishing role of TRPand thermogenesis, which can lead to calorie channel modulating dietary agents in obesity.consumption and prevention of obesity. Commondietary spices like chilli pepper, black pepper,Research in Progress:clove, garlic, cinnamon and their constituents Based on our PCR data multiple TRP channel(capsaicin, piperine, eugenol, allicin, genes are expressed in mouse 3T3-L1cinnamaldehyde, menthol) can activate the TRP preadipocytes (high to moderate expression:channels. In this proposal, using the TRP channel TRPP2 (PKD2), TRPC1, TRPV2, TRPM2,receptor system we intend to come up with dietary TRPV4 TRPV1, TRPV3, TRPV6, TRPC4; lowconstituents that can modulate the molecular expression: TRPA1, TRPC6, and TRPM8) andmechanism associated with the process of adipocytes (high to moderate expression: TRPP2adipogenesis, adipose tissue related hormonal (PKD2), TRPV2, TRPC1, TRPV4, TRPM2; lowsecretion and release of pro-inflammatory expression: TRPA1, TRPV3, TRPV6, TRPC4,mediators and lipolysis.TRPV1, TRPC6, and TRPM8) (Table 1). TRPV1,TRPV3, TRPM8, TRPC4, TRPC6 wereResearch Objectives:differential expressed (multiple fold higher inpreadipocytes than adipocytes) (Figure 2).l Determination of expression and function ofTRP channels in commercially availableFurther, these channels are also expressed inmouse preadipocytes cell lines (3T3-L1),murine white adipose tissue (WAT) (high tomoderate expression: TRPP2 (PKD2), TRPV2,human preadipocytes (HPAd) and adipocytesTRPC6, TRPC1, TRPV4, TRPM2, TRPV3,(HAd) cells.TRPC4; low expression: TRPV1, TRPV6,l In-vitro characterization of the molecular TRPM8 and TRPA1] and brown adipose tissuebasis of adipogenesis using mouse/human (BAT) [high to moderate expression: TRPP2gene microarray chip, mouse/human (PKD2), TRPV2, TRPC6, TRPC1, TRPV4,adipogenesis and obesity PCR arrays. TRPM2, TRPV3, TRPC4, TRPV6; low expression:Determination of effect of TRP channel TRPV1, TRPM8, and TRPA1) (Table 1). TRPV4,activation on adipogenesis process in TRPV6 and TRPC6 were differentially expressedmouse/human adipocytes cells.in murine WAT and BAT (Figure 2).32

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTETable 1: Summary of TRP channel expression in (A) undifferentiated (preadipocytes) anddifferentiated (adipocytes) 3T3-L1 cells (B) mouse white (WAT) and brown (BAT) adipose tissueTRP GeneMouse 3T3-L1 cell line Mouse adipose tissuePreadipocyte Adipocyte WAT BATV1 ++ + ++ ++V2 ++ ++ +++ +++V3 ++ + ++ ++V4 ++ ++ +++ ++V6 ++ + +++ ++M2 ++ ++ ++ ++M8 ++ + + +A1 ++ + + +C1 +++ ++ ++ ++C4 ++ + ++ ++C6 + + +++ +++P2 +++ +++ +++ +++GAPDH ++++ ++++ ++++ ++++(++++ = very high expression (ct values= 30).Figure 1: Differential expression of TRP channel in (A) undifferentiated (preadipocytes) and differentiated(adipocytes) 3T3-L1 cells relative to GAPDH (House Keeping Gene, HKG) (B) mouse white (WAT) andbrown adipose tissue (BAT) relative to TRPP2 (HKG). * significant with p value =< 0.01; ** significant with pvalue =< 0.03.33

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE4.1.3 Characterization of arabinoxylans in regulating their biological activities will becereal grainsevaluated using in-vitro and in-vivo model, whichwill further allow us to understand the structurefunctionrelationship of the AXs in Indian milletPrincipal Investigator:Koushik Mazumdervarieties with respect to their biological activities.Introduction:Compositional sugar analysis and structuralcharacterization of the arabinoxylanpolysaccharides and oligosaccharides usingvarious chemical and modern analyticalmethods (GC-MS, HPLC, MALDI-TOFand ESI-MS/MS) to establish structure-function relationship.Cereals, the staple foods for millions of peopleacross the world, are the chief source of solubledietary fibers. Several epidemiological studieshave clearly demonstrated that increasedconsumption of whole grain cereals and solubledietary fibers has been associated with a reducedrisk of many life style disorders such as type-2diabetes and obesity. In cereal grains, AXs are themajor non-starchy polysaccharides whichconstitute the cell walls residues in cereal grainsand make up an important physicochemical andphysiological property beneficial to the health ofconsumers and for this reason, their use as foodingredients has increased rapidly. Although AXsshare the same basic chemical structure, but thepattern and degree of substitution of arabinosealong the xylan backbone vary with cerealsources. Therefore, the AXs exhibits a great dealof structural heterogeneity and variability withrespect to molecular weight mass, xylose toarabinose ratio and branching pattern,distribution of arabinose residues and substitutionwith glucuronic acid/4-O-methyl glucuronicacids.There is no documented report about thecomparative study of arabinoxylan poly andoligo-saccharides from Indian millet varietiessuch as finger millet, kodo millet, branyard milletand foxtail millet. Hence in the present study, thevariability in the structures of the AXs fromvarious Indian millet varieties and their role inResearch Objectives:The present study aims to elucidate comparativestatement profile of AXs from Indian milletvarieties and its effect on their biologicalactivities.llIsolation and purification of the arabinoxylanpolysaccharides and oligosaccharides fromthe bran of Indian variety millets.Long Term Objective:Development of nutraceutical health foods basedon dietary fibers enriched with Axs.Research in Progress:In our preliminary studies, we havestandardized the extraction protocol for theisolation of AXs from Finger millet bran and thecompositional analysis of the polysaccharideshave been carried out (GC, GC-MS analysis) asmonomeric alditol acetate derivative with inositolas internal standard (Figure 2). The compositionalanalysis showed the presence of arabinoxylan asmajor constituent (~90%) together with starch andgalactomannan as minor component.Figure 2: GC chromatogram of the monomeric sugar residues of extracted arbinoxylan fraction from fingermillet bran. (Ara: Arabinose, Xyl: Xylose, Inos: Inositol, Gal: Galactose, Glc: Glucose, Man: Mannose).34

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE5. COMPUTATIONAL BIOLOGY APPROACHES FORMARKER AND GENE DISCOVERY FOR NUTRITION ANDPROCESSING TRAITS IN GENOMES OF FOOD CROPS5.1 Development of advanced algorithms, Research Objectives:databases, tools and pipeline for data miningland comparative analysis of food cropgenomes, transcriptome and small RNA basedregulation.lCoordinator:Rakesh TuliInvestigators:Shrikant Subhash MantriJoy K. RoyProject Scientists:Shailesh SharmaVandana MishralllTranscriptome sequencing, assembly andannotation for contrasting wheat varieties.Digital expression profiling and microarraydatamining for identification of SingleFeature Polymorphism (SFP) in Indiancultivars and annotation of hypotheticalproteins.SNP database development in wheat.Comparative genomics and related datamining.Small RNA analysis pipeline development.Research Fellows:l Microarray datamining for SFP.Gourab Dasl Repository of germplasm.Anuradha SinghLong Term Objectives:Introduction:lBread wheat, Triticum aestivum, is anFramework map development for wheatgenome (assembly and annotation).allohexaploid species. Its genome is composed ofl Identification of genes involved in highthe three distinct ancestral genomes A, B and D.accumulation of micronutrients.The polyploid nature of the wheat genometogether with its large size and high percentage of l Genome wide association analysis forrepeat elements makes it extremely complex and co-rrelating quality and processing relatedcomputationally intensive to analyze ever traits with the identified SNP's.increasing transcriptome and genome data. With5.1.1 SNP database development in hexaploidthe advent of next-generation sequencingwheattechnology it is now possible to generate wholegenome/transcriptome sequences for a number of Research in Progress:wheat genome. We are interested to harness the Datamining of non-synonymous mutations orpublic domain data of wheat for different projects Single Amino acid Polymorphisms (SAPs) fromon crop improvement in terms of nutritional and ESTs and homoeologous SNPs from chromosomeprocessing quality. Additionally, NABI has 1 of Triticum aestivum, resulted into theinitiated experiments to generate transcriptome identification putative 35395 and 1817 SNPssequences for Indian wheat varities showing respectively.contrasting characters. Bioinformatics Labortarywhich has the high performance computing nsSNP mining in ESTs from dbEST:infrastructure is a gateway for NABI scientists to Non-synonymous deleterious mutations are fairlyuse computational methods and databases tocommon in crop genomes. Statistical analysis ofdeepen and integrate their research.homologous sequence from multiple genomes can35

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEamino acid across species suggests that the non-synonymous change (indicated by the red 'G')observed in maize is likely to be deleterious.Synonymous changes are shown in black.identify amino acid changes that are likely to bedisadvantageous. Figure 1 shows a hypotheticalalignment of coding sequence from multiple grassspecies. The conserved nature of the histidineFigure 1: Hypothetical alignment of coding sequence from multiple grass species.Non-synonymous change indicated in red, synonymous change indicated in black.Highly putative nsSNPs in hexaploid wheat ESTs were found through the following steps (Table 1-7):1. dbEST records were parsed using custom perl script and multifasta containing all EST sequenceswas generated.Table 1: dbEST summary (updated February 1, 2012).No. of Triticumaestivum ESTsNo. ofcultivarsNo. of tissuesNo. of developmentalstages1073845 101 108 1322. These EST sequences were clustered using BLAST algorithm.Table 2: EST clustering results. Current release March 19, 2012contains 36692 unigenes.No. of Triticumaestivum probe setsNo. of unigenescoveredNo. of probe setshaving no clusterinfoNo. of probe setshaving cluster depth> 5 sequences61115 24697 123 463723. SNPs were determined using autosnpV2 script.Table 3: Overall SNPs summary.No. ofProbesetsSNPsSynonymous SNPsSynonymousSNPs inCDSSynonymousSNPs in flankingCoding regionproteinsubstitutionsSNPs indomains20699 161188 94654 67478 27176 35395 248636

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE4. nsSNPs were extracted from the overall SNP result.Table 4: nsSNP statistics.Total no. of nsSNPsNo. of probe setsinvolvedInvolved probe setswith annotationNo. of nsSNPs withannotation35395 12441 12409 352335. SNPs in transcription factors were identified.Table 5: Summary of SNPs in Transcription Factors.No. of probe sets Synonymous SNPs No. of nsSNPs SNPs in domains188 776 335 1066. SNPs in pathways were also determined.Table 6: Summary of mutations in pathways.No. of pathwaysNo. of probesets involvedNo. of enzymesNo. ofsynonymousSNPsNo. of nonsynonymoussubstitutions147 2157 521 13442 45567. SNPs in starch & sucrose metabolism pathway.Table 7: SNP summary in Starch Metabolism Pathway.Pathway IDPath: map00500No. of enzymesinvolvedNo. of probe setsinvolvedNo. ofsynonymousSNPsNo. of nonsynonymoussubstitutions42 214 1840 525nsSNPs in remaining 11995 unigenes from Unigene database of NCBI is being detected.Homoeologous SNPs in chromosome1:According to SNP nomenclature, a homoeologousSNP exists if there is a difference between thethree homoeologous genomes (A, B and D) in asingle individual. Varieties 1 and 2 possess thesame homoeologous SNP and have thecharacteristic of a single individual.Figure 2: Depiction of homoeologus variation between A, B and D genome in wheat.37

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEHomoeologous SNP mining was carried outthrough the following steps (Table 8-11):1) SRA reads were retrieved from NCBI SRAdatabase and multifasta was generated.2) Reads were assembled by CLC Assembly Cellsoftware with default parameters.3) Resultant contigs were filtered to get thehomoeologs using cut-off value of minimum5 reads from each of the 3 genomes (A,B,D)producing 105 homoeologs out of which 90homoeologs have the polymorphism whichis determined by autosnpV1 script.4) SNPs in the regulatory region were identifiedusing cis regulatory motifs in PLACEdatabase.Table 8: Assembly results of hexaploid wheat chromosomeCLC versionNo. of SRAreadsSingleton Assemble reads Total no. ofcontigs4 beta 1072765 6126635 4601040 8854313.2 1072765 2850260 7877415 821609Table 9: Assembled contig summary.Total no. ofcontigsNo. of contigswith reads from1 genomeNo. of contigswith reads from2 genomesTable 10: Homoeologous SNPs summary.No. of contigshaving readsfrom allgenomesNo. of contigswith no readinformation885431 284159 71530 47205 482537821609 443337 130377 81295 166600Both 3.2 and 4 beta versions were used to compare the number of identified SNPs.No. of contigsReads from AgenomeReads from BgenomeReads from DgenomeTotal no. ofSNPs90 4254 4810 1182 1817Table 11: PLACE database screening report.No. of contigsTotal no. ofregulatory SNPsNo. of contigswith noregulatory SNPsNo. of annotatedcontigs withregulatory SNPsNo. of annotatedregulatory SNPs90 369 31 40 2505) Calculation of GC content for all the genomes from chromosome 1 (Table 12).Table 12: Nucleotide base summary and GC contents of 3 genomes.Features Genome A Genome B Genome DGC content 0.4317 0.4368 0.443038

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE5.1.2 Wheat genechip microarray data mining out of the 61127 probe sets of wheat.for identification of SFPs in Indian cultivars and Optimization of script is under progress.annotation of hypothetical proteinsIntroduction:5.1.3 Development of small RNA analysispipelineSNPs and insertion or deletions of one or morenucleotides (indels) are DNA polymorphisms thatcan affect hybridization of DNA or cRNA to aprobe on an array. The Affymetrix Gene Chiparrays are suitable to detect such variationsbecause each gene is represented by a set of eleven25-bp probes that are sensitive to target mismatchowing to their short sequence. A target sequencethat matches the sequence of a probe binds withmuch greater affinity than one with a mismatchsequence. The resulting difference inhybridization intensity between two genotypes foran individual probe is called a SFP, where a featurerefers to a probe in the array. A SFP may be causedby a SNP, a multiple nucleotide polymorphism, oran indel.Research Objective:To predict SFPs in the genomes of the followingwheat varieties: C-306, LOK1, Sonalika andWH291, using microarray data.Research in Progress:1. An algorithm was written for predictingSFPs using the normalized expression signalvalues of the four wheat varieties.2. Standard deviation values were computedusing the normalized signal intensity valuesof perfect match probes of a probeset for allthe four wheat varieties 7 DAA,14 DAA and 28 DAA.Introduction:miRNAs are small and endogenous RNAs thatregulate endogenous genes or gene expression.miRNAs are important for implementingdevelopmental programs in animals and plants.Our research focus is on plant miRNA predictionand its target prediction (miRNA). In plants, these~18-24 nt RNAs are processed from stem-loopregions of long primary transcripts by a dicer-likeenzyme and are loaded into silencing complexes,where they generally direct cleavage ofcomplementary mRNAs. NABI has initiatedexperiments to generate small RNA sequencesusing Illumina platform for RNA isolated fromcontrasting samples of wheat, custard apple, litchiand mango to understand small RNA regulationand identify novel regulatory small RNA. Longterm goal under this component is to reconstructgene regulatory networks using multi-dimensiondata. Small RNA prediction and its targetprediction will be done using in-house developedpipeline of open source prediction tools andcustomized scripts. Bench marking of differentsoftwares (in the pipeline) is in progress usingdatasets available in public domain.Research Objectives:lBench-marking of miRNA prediction andmiRNA-target prediction tools.3.l Development of small RNA analysisFour average standard deviation values werePipeline.used as reference values to predict SFP. SFPis predicted if the standard deviation valuesl Development of in-house small RNAof a probe of a probeset are less than or equal database.to the minimum of the four average standarddeviation values.l Comparative analysis of small RNA4. Python script was written based on the aboveregulation.written statistical logic for predicting SFPs l Reconstruction of gene regulatory networks.39

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEEMERGING AREAS1. Probing B-ZIP transcription factors role in 2. Developing carbohydrate and lipidroot and seed development in plants: Use of a derivative based nano molecular vehicles fordesigned dominant negative proteinbioavailability of micronutrientsInvestigator: Vikas RishiInvestigator:Nitin K. SinghalThere are approximately 72 B-ZIP transcriptionfactors in Arabidopsis and a number of these areinvolved in root and seed development. The B-ZIPmotif is a long bipartite α-helix. The C-terminalhalf is an amphipathic α-helix that dimerizes toform a parallel dimeric coiled-coil termed asMicronutrient deficiency such as iron deficiencymay arise from inadequate intake and impairedabsorption leading to anaemia, reduce cognitivedevelopment and lower work capacity. Althoughiron deficiency therapy with oral ferrous salts iswidespread, the bioavailability of iron from suchsalts is poor. Anti-nutrient components, such asphytate in whole grain and bran, oxalic acid inspinach, phosphates in practically all foods, andtannins from black tea and coffee may decrease+2the bioavailability of ferrous iron Fe by forminginsoluble complexes .FeSO 4 is water soluble compound, but is easily+ 3oxidized to low soluble Fe form after contactFigure 1: Design of dominant negative to B-ZIP:A-ZIP. with oxygen. This may often cause unacceptablechange in the color and taste of foods. Moreover,leucine zipper. The N-terminal half is a basic +2Fe frequently causes gastrointestinal side effects.region that binds sequence specific DNA. AIt is highly desirable to find a better alternativedominant negative (DN) protein called A-ZIP inwhich the basic DNA binding region of a B-ZIPiron source that can overcome the gastrointestinaltranscription factor (TF) is replaced by a rationally side effects of iron. Iron oxide/hydroxidedesigned acidic extension has been used nanoparticles with neutral and hydrophilicsuccessfully in mammalian systems to study the carbohydrate nanosize shell can be used asrole of these TFs in growth, development and alternatives to ferrous salts. In these formulationsdifferentiation (Figure 1). These DN proteins gastrointestinal side effects are rare becauseheterodimerize with the wild type proteins and3+hundreds of Fe atoms are safely packed ininhibit their function by not allowing them to bind nanoscaled cores surrounded by the solubilizingto promoter of a gene. DN is biologically active shell. There are some evidences that particles inreagent because a heterodimer is more stable thanthe range of nanometre show different propertiesa B-ZIP homodimer bound to DNA. This strategyas compared to larger particles. This and the factwill be used to study root and seed development inthat very large surface area of nanoparticles allowsArabidopsis. Use of dominant negative forsilencing genes has advantage over other RNAiron to be much more bioavailable, suggesting thebased technologies like siRNA and miRNA. potential of designing and producing particles ofThese designed dominant negatives can inhibit the nano dimension. Nanoparticle based deliveryfunctions of all the TFs belonging to the same systems can boost iron bioavailability and mayfamily, thus overcoming the problem of play a vital role in food applications in the nearredundancy, a phenomenon common in biological future for treating micronutrient deficiencies.systems.40

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTESYNERGY THROUGH COLLABORATIONSAND NETWORKING1. NABI and NIPER signed a MOU on 18.02.2012 to undertake joint research work in the area ofmutual interest besides imparting training to staff, students and technical personnel within the areasof cooperation.2. NABI is in advanced stage of signing MOUs with the following Institutes/Universities:i. Punjab Agricultural University, Ludhiana.ii. Punjab Technical University, Kapurthalaiii. Birla Institute of Technology, Pilani.41

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTENEWLY JOINED FACULTY43

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE(BRIEF PROFILES)Name: Dr. Vikas RishiDate of Joining: 01-03-2012Designation: Scientist EArea of Research Interest: Epigenetics, DNA methylation and gene regulation,transcription factors dimerization and DNA binding specificity, proteinengineering in deciphering biological functions of B-ZIP transcription factors.Past Appointments:1. Visiting Scientist (2001-2006): National Cancer Institute, National Institutes of Health, Bethesda,USA.2. Research Fellow (FTE position) (2006-2009): National Cancer Institute, National Institutes ofHealth, Bethesda, USA.Major Publications:1. Warren CL, Zhao J, Glass K, Rishi V, Ansari AZ, Vinson C (2012). Fabrication of duplex DNAmicroarrays incorporating methyl-5-cytosine. Lab Chip. 12 (2): 376-80.2. Eric RD, Chatterjee R, Zhao J, Rishi V, Vinson C (2012). Dominant negative induces degradation ofB-ZIP proteins. Biochem Biophys Res Commun. (in press).3. Rishi V, Bhattacharya P, Chatterjee R, Rozenberg J, Zhao J, Glass K, Fitzgerald P and Vinson C(2010). CpG methylation of half-CRE sequences creates C/EBP alpha binding sites that activatesome tissue-specific genes. Proc Natl Acad Sci U S A.107 (47): 20311-6.4. Rishi V, Oh WJ, Heyerdahl SL, Zhao J, Scudiero D, Shoemaker RH and Vinson C (2010).Arylstibonic acids that inhibit the DNA binding of five B-ZIP dimers. J Struct Biol. 170 (2): 216-25.5. Rishi V, Gal J, Krylov D, Fridriksson J, Boysen MS, Mandrup S, Vinson C (2004). SREBP-1dimerization specificity maps to both the helix-loop-helix and leucine zipper domains: use of adominant negative. J Biol Chem. 279 (12): 11863-74.45

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. Ajay K. PandeyDate of Joining: 14-11-2011Designation: Scientist DArea of Research Interest: Functional genomics approaches in wheat andlegumes crop plants to understand the linkage between the gene function and traitdevelopment. Metabolic engineering for trait development.Past Appointments:1. Post Doctoral Research Associate (2008-2011): Department of Plant Pathology, 351 Bessey Hall,Iowa State University, Ames, Iowa 50011.2. Post Doctoral Fellow (2004-2008): Department of Biological Sciences, University of Alabama,Huntsville, AL-35899.3. Post Doctoral Fellow (2003-2004): Institute of Plant and Microbial Biology, Academia Sinica,Taipei, Taiwan.Major Publications:1. Pandey AK, Yang C, Zhang C, Graham MA, Hill JH, Pedley KF and Whitham S (2011). Functionalanalysis of the genes that contribute to Rpp2-mediated defense against Asian soybean rust.Molecular Plant Microbe Interactions. 24: 196-204.2. Pandey AK, Jain P, Podila GK, Tudzynski B and Davis MR (2009). Cold induced Botrytis cinereaenolase (BcEnol-1) functions as a transcriptional regulator and is controlled by CAMP. MolecularGenetics and Genomics. 281: 135-146.3. Cseke LJ, Ravinder N, Pandey AK and Podila GK (2007). Identification of PTM5 proteininteraction partners, a MADS-box gene involved in aspen tree vegetative development. Gene. 391:209-222.4. Huang HEn, Ger MJ, Ying CC, Pandey AK, Yip MK, Lin MK, Chou HK and Feng TY (2007).Disease resistance to bacterial pathogens affected by the amount of ferredoxin-I protein in plants.Molecular Plant Pathology. 8: 129-137.5. Pandey AK, Ger MJ, Huang HEn, Yip MK, Lin MK, Zeng J and Feng TY (2005). Expression of thehypersensitive response-assisting protein in Arabidopsis results in harpin dependent hypersensitivecell death in response to Erwinia carotovora. Plant Molecular Biology. 59: 771-780.46

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. Kanthi K. KiranDate of Joining: 02-09-2011Designation: Scientist CArea of Research Interest: Dietary constituents for the regulation of metabolicsyndrome- nutrigenomics study, functional foods (probiotics, prebiotics andsynbiotics), bacterial exopolysaccharides.Past Appointments:1. Post Doctoral Fellow (2009-2011): Lund University, Lund, Sweden2. Lecturer (2002-2003): TSR & TBK Degree College and P.G Centre, Gajuwaka, VisakhapatnamMajor Publications:1. Kondepudi KK, Ambalam P, Wadström T and Ljungh A (2012). Prebiotic-non-digestibleoligosaccharides preference of probiotic bifidobacteria and antimicrobial activity againstclostridium difficile. Anaerobe. 18 (5): 489-97.2. Ambalam P, Kondepudi KK, Wadström T and Ljungh A (2012). Bile stimulates cell surfacehydrophobicity, congo red binding and biofilm formation of Lactobacillus strains. FEMS MicrobiolLett. 333 (1): 10–19.3. Kondepudi KK and Chandra TS (2011). Isolation of a moderately halophilic and amylolyticbacterium from saline soil of a salt manufacturing industry. J. Pure Appl Microbiol. 5 (2): 545-552.4. Kondepudi KK and Chandra TS (2011). Identification of osmolytes from a moderately halophilicamylolytic Bacillus sp. strain TSCVKK. Eur. J. Expt. Biol. 1 (1): 113-121.5. Kondepudi KK and Chandra TS (2008). Production of surfactant and detergent-stable, halophilicand alkalitolerant alpha-amylase by a moderately halophilic Bacillus sp. Strain TSCVKK. Appl.Microbiol. Biotechnol. 77: 1023-1031.47

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. Mahendra BishnoiDate of Joining: 16-12-2011Designation: Scientist CArea of Research Interest: Receptor pharmacology (metabolic disorders andnervous system disorders)Past Appointments:1. Research Associate (2009-2011): Department of Pharmacology, SouthernIllinois University- School of Medicine, Springfield, Illinois, USA.2. Post Doctoral Associate (2009-2009): Department of Psychiatry, McKnight Brain Institute,University of Florida, Gainesville, Florida, USA.3. Assistant Professor (Temporary) (2007-2008): Department of Pharmacology, University Instituteof Pharmaceutical Sciences (UIPS), Panjab University, Chandigarh.4. Research Assistant (2004-2005): Pharmacology Division of Bioresearch Division, Sun PharmaAdvanced Research Centre (SPARC), Vadodara, Gujarat.Major Publications:1. Cao DS, Zhong L, Hsieh TH, Abooj M, Bishnoi M, Hughes L and Premkumar LS (2012).Expression of TRPA1 and its role in insulin release from rat pancreatic beta cells, PLoS ONE. 7(5):e38005.2. Bishnoi M, Bosgraaf CS, Abooj M and Premkumar LS (2011). Involvement of TRPV1 andinflammation in STZ-induced early thermal hyperalgesia is independent of glycemic state of rats.Molecular Pain. 7: 52.3. Bishnoi M, Bosgraaf CS and Premkumar LS (2011). Preservation of acute pain and efferentfunctions following intrathecal resiniferatoxin-induced analgesia. Journal of Pain. 12: 991-1003.4. Bishnoi M, Chopra K and Kulkarni SK (2009). Co-administration of nitric oxide (NO) donorsprevents haloperidol-induced orofacial dyskinesia, oxidative damage and change in striataldopamine levels. Pharmacol Biochem Behav. 91: 423-429.5. Bishnoi M, Chopra K and Kulkarni SK (2008). Modulatory effect of neurosteroids in haloperidolinducedorofacial dyskinesia: An animal model of Tardive Dyskinesia, Psychopharmacology.196: 243-254.48

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. Koushik MazumderDate of Joining: 01-02-2012Designation: Scientist CArea of Research Interest: Carbohydrate chemistry, glycobiology.Past Appointment:1. Post Doctoral Research Associate (2008-2011): Complex CarbohydrateResearch Centre, University of Georgia, 315 Riverbend Road, Athens,Georgia 30602, USA.Major Publications:1. Correia MAS, Mazumder K, Brás JLA, Firbank SJ, Zhu Y, Lewis RJ, York WS, Fontes CMGA andGilbert HJ (2011). The structure and function of an arabinoxylan-specific xylanase. Journal ofBiological Chemistry. 286: 22510-22520.2. Mazumder K and York WS (2010). Structural analysis of AXs isolated from ball milled switchgrassbiomass. Carbohydrate Research. 345: 2183-2193.3. Mazumder K, Choudhury BP, Nair GB and Sen AK (2008). Identification of a novel sugar 5, 7-diacetamido-8-amino-3,5,7,8,9-pentadeoxy-D-glycero-D-galacto-non-2-ulosonic acid present inthe lipooligosaccharide of Vibrio parahaemolyticus O3:K6. Glycoconjugate Journal. 25: 345–354.4. Zhu X, Pattathil S, Mazumder K, Brehm A, Hahn MG, Dinesh-Kumar SP and Joshi CP (2010).Virus induced gene silencing offers a functional genomics platform for studying plant cell wallformation. Molecular Plant. 3: 818-833.5. Kulkarni AR, Peña MJ, Avci U, Mazumder K, Urbanowicz B, Yin Y, O'Neill MA, Roberts AW,Hahn MG, Xu Y, Darvill AG and York WS (2011). The ability of land plants to synthesizeglucuronoxylans predates the evolution of tracheophytes. Glycobiology. 22: 439-451.49

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. Nitin K. SinghalDate of Joining: 02-03-2012Designation: Scientist CArea of Research Interest: Synthetic nano-biochemistry, liposome based genedelivery, fabrication of nano / microstructures, cellular assays, organic synthesisof small molecules, biosensors, biomarker based disease diagnostic.Past Appointments:1. Post Doctoral Fellow (2008-2009): Pennsylvania State University, State College, USA2. Post Doctoral Fellow (2009-2012): Seoul National University, Seoul, South KoreaMajor Publications:1. Kumar A, Ramanujam B, Singhal NK, Mitra A and Chebrolu PR (2010). Interaction of aromaticimino glycoconjugates with jacalin: experimental and computational docking studies CarbohydrateRes. 345: 2491-2498.2. Amit K, Singhal NK, Nagender RR, Siva KN and Chebrolu PR (2009). C1- and C2-Imino-basedglycoconjugates: inhibition aspects towards glycosidase activities isolated from soybean and jackbean. Glycoconjugates Journal. 26: 495-510.3. Singhal NK, Mitra A, Ramanujam B and Chebrolu PR (2009). Variation in the sensitivity and2+selectivity of M as a function of the stereochemistry of the carbohydrate: Selectivity differences2+between galactosyl and ribosyl derivatives towards Cu attributed to the structural differences.Dalton Tran. 39: 8432-8442.4. Singhal NK, Ramanujam B, Mariappanadar V and Rao CP (2006). Carbohydrate-based switch-onmolecular sensor for cu (ii) in buffer: Absorption and fluorescence study of the selective recognitionof cu (ii) ions by galactosyl derivatives in HEPES buffer. Organic Letters. 8: 3525-3528.5. Ahuja R, Singhal NK, Ramanujam B, Ravikumar M and Rao CP (2007). Experimental andcomputational studies of the recognition of amino acids by galactosyl-imine and -amine derivatives:An attempt to understand the lectin-carbohydrate interactions. Journal of Organic Chemistry.72: 3430-3442.50

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. Shailesh SharmaDate of Joining: 02-01-2012Designation: Project ScientistArea of Research Interest: Detecting single feature polymorphisms in Triticumaestivum using affymetrix arrays. Transcript annotation of Triticum aestivumobtained from affymetrix.Past Appointment:1. Post Doctoral Scientist (2010 – 2011): USA - India joint research training program. This programwas hosted by the Seattle Biomedical Research Institute at the University of Washington, USA and atThe International Centre for Genetic engineering and Biotechnology, New Delhi. Research programwas funded by the Fogarty International Center of the National Institutes of Health, USA.Major Publications:1. Sharma S, Cavallaro G and Rosato A (2010). A systematic investigation of multiheme c-typecytochromes in prokaryotes. J. Biol. Inorg. Chem. 15: 559-571.2. Sharma S and Rosato A (2009). Role of the N-terminal tail of metal-transporting P (1B)-typeATPases from genome-wide analysis and molecular dynamics simulations. J. Chem. Inf. Model. 49:76-83.51

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. NishimaDate of Joining: 04-01-2012Designation: Project ScientistAreas of Research Interest: Nanobiotechnology and its applications,microcantilever based detection of biomolecules, polypeptides basedmicroelectronics, chemical modification of haptens and physicochemicalcharacterization, natural product chemistryPast Appointment:1. Post Doctoral Research Fellow (2010-2011): Nanyang Technological University, SingaporeMajor Publications:1. Wangoo N, Shekhawat G, Jin-Song Wu, Suri CR, Bhasin KK and Dravid V (2012). Green synthesisand characterization of size tunable silica capped gold core-shell nanoparticles. Journal ofNanoparticle Research (In Press).2. Wangoo N, Kaushal J, Bhsasin KK, Mehtaand SK and Suri C (2010). Zeta potential basedcolorimetric immunoassay for the direct detection of diabetic marker HbA1c using gold nanoprobes.Chemical Communications. 46: 5755-5757.3. Wangoo N, Bhasin KK, Mehta SK and Suri CR (2008). Synthesis and capping of water-dispersedgold nanoparticles by an amino acid: Bioconjugation and binding studies. Journal of Colloid andInterface Science. 323: 247-254.4. Wangoo N, Bhasin KK, Boro R and Suri CR (2008). Facile synthesis and functionalization of watersolublegold nanoparticles for a bioprobe. Analytica Chimica Acta. 610: 142-148.52

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Dr. Sudhir P. SinghDate of Joining: 16-01-2012Designation: Project ScientistArea of Research Interest: Molecular biology of seed development in wheatwith the objective to identify bottlenecks by preventing iron translocation fromouter bran layers to endosperm in bioavailable form. To identify the candidategenes for inducing seedlessness in fruit crops.Past Appointment:1. Research Associate (2010-2011): National Agri-Food Biotechnology Institute, Mohali, PunjabMajor Publications:1. Kumar J, Singh SP, Kumar J and Tuli R (2012). A novel mastrevirus infecting wheat in India.Archives of Virology. 157(10): 2031-2034. ISSN: 0304-8608.2. Kumar J, Kumar A, Singh SP, Roy JK, Lalit A, Parmar D, Sharma NC and Tuli R (2012). First reportof leaf curl virus infecting okra in India. New Disease Reports. 25: 9. ISSN No. 2044-0588.3. Singh SP, Pandey T, Srivastava R, Verma PC, Singh PK, Tuli R and Sawant SV (2010). BECLIN 1from Arabidopsis thaliana, under the generic control of regulated expression systems, a strategy fordeveloping male sterile plants. Plant Biotechnology Journal, 8(9): 1005–1022. ISSN: 1467-7652.International Patent (Published): Sawant SV, Tuli R and Singh SP. Method for producing malesterile plants. PCT WO/2010/061276; A gene for inducing male sterility. India: 2697/DEL/2008;USA: US2011289631; European Union: EP2350290; China: CN102257143; Australia:AU2009321261.4. Lodhi N, Ranjan A, Singh M, Srivastava R, Singh SP, Chaturvedi CP, Ansari SA, Sawant SV andTuli R (2008). Interactions between upstream and core promoter sequences determine geneexpression and nucleosome positioning in tobacco PR-1a promoter. Biochimicaet Biophysica Acta(BBA). 1779 (10): 634-644. ISSN: 1874-9399.5. Singh RK, Singh SP and Singh SB (2005). Correlation and path analysis in sugarcane ratoon. SugarTech. 7(4): 176-178. ISSN: 0972-1525.Extramural Grants Received: Title: A novel strategy for developing scion plants of desired phenotype,by using RNAi delivering rootstock. Sponsoring Agency: Science and Engineering Research Board,Department of Science and Technology (DST).53

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEName: Ms. Vandana MishraDate of Joining: 23-01-2012Designation: Project ScientistArea of Research Interest: Bioinformatics: Development of genomicsresources for grain crops.Past Appointments:1. Junior Research Fellow (Project- Next generation sequencing) (2011-2012): Plant Immunity Laboratory, National Institute of Plant Genome Research, JNU campus, NewDelhi.2. Application Specialist (Biotechnology instruments/bioinformatics software) (2010-2011):NSW Biotech Pvt. Ltd, New Delhi.3. Junior Research Fellow (Project- Structural bioinformatics) (2009-2010): Department ofBiochemistry, Delhi University, South campus, New Delhi.4. Bioinformatician (2008-2009): Infovalley Biosystems (India) Pvt. Ltd, Bangalore.Major Publications: Nil54

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTERESEARCH PUBLICATIONS55

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEI. Publication based on research initiated at NABI:1. Kumar J, Singh SP, Kumar J and Tuli R (2012). A novel mastrevirus infecting wheat in India.Archives of Virology. 157(10): 2031-2034.II.Publications based on work done by faculties or initiated at their earlier institutes:20121. Bishnoi M and Boparai RK (2012). An animal model to study the molecularbasis of tardive dyskinesia. Methods Mol Biol. 829: 193-201.2. Bruijnzeel AW, Ford J, Rogers JA, Scheick S, Ji Y, Bishnoi M and Alexander JC (2012). Blockade ofCRF1 receptors in the central nucleus of the amygdala attenuates the dysphoria associated withnicotine withdrawal in rats. Pharmacol Biochem Behav. 101(1): 62-8.3. Chitale VS, Tripathi P, Behera MD, Behera SK and Tuli R ( 2012 ). On the relationships amongdiversity, productivity and climate from an Indian tropical ecosystem: a preliminary investigation.Biodiversity and Conservation. 21: 1177-1197.4. Jena SN, Srivastava A, Rai KM, Ranjan A, Singh SK, Tarannum N, Srivastava M, Bag SK, Mantri S,Asif MH, Yadav HK, Tuli R and Sawant SV (2012). Development and characterization of genomicand expressed SSRs for levant cotton (Gossypium herbaceum L.). Theor Appl Genet. 124: 565-576.5. Kumar J, Kumar A, Singh SP, Roy JK, Lalit A, Parmar D, Sharma NC and Tuli R (2012). Firstreport of leaf curl virus infecting Okra in India. New Disease Report (earlier BSPP Plant Pathology),25: 9, ISSN No 2044-0588.6. Madnala R, Gupta V, Singh PK and Tuli R (2012). Development of chloroplast transformationvectors, and a new target region in the tobacco plastid genome. Plant Biotechnol Reports. 6: 77-87.7. Ranjan A, Nigam D, Asif MH, Singh R, Ranjan S, Mantri S, Pandey N, Trivedi I, Jena SN, Tuli R,Pathre U and Sawant S (2012 ). Genome wide expression profiling of two accessions of Gossypiumherbaceum L. in response to drought. BMC Genomics. 13: 94-99.8. Soumit K, Behera , Mishra AK , Sahu N , Kumar A, Singh N, Kumar A, Bajpai O, Chaudha L B,Khare PB and Tuli R ( 2012 ). The study of microclimate in response to different plant communityassociation in tropical moist deciduous forest from northern India. Biodiversity and Conservation.21: 1159-1176.9. Singh SP, Singh Z and Swinny EE (2012). Climacteric level during fruit ripening influences lipidperoxidation and enzymatic and non-enzymatic antioxidative systems in Japanese plums (Prunussalicina Lindell). Postharvest Biology and Technology. 65: 22-32.10. Tiwari S and Tuli R (2012). Optimization of factors for efficient recovery of transgenic peanut(Arachis hypogaea L.) Plant Cell Tissue and Organ Culture. 109: 111–121.11. Upadhyay SK, Singh S, Chandrashekar K, Tuli R and Singh PK (2012). Compatibility of garlic(Allium sativum L.) leaf agglutinin and CryAc -endotoxin for gene pyramiding. AppliedMicrobiol & Biotechnol. 93: 2365-2375.12. Warren CL, Zhao J, Glass K, Rishi V, Ansari AZ and Vinson C (2012). Fabrication of duplex DNAmicroarrays incorporating methyl-5-cytosine. Lab Chip. 12 (2): 376-80.57

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTE20111. Bishnoi M, Bosgraaf CA, Abooj M and Premkumar LS (2011). Involvement of TRPV1 andinflammation in STZ-induced early thermal hyperalgesia is independent of glycemic state of rats.Molecular Pain. 7: 52.2. Bishnoi M, Bosgraaf CS and Premkumar LS (2011). Preservation of acute pain and Efferentfunctions following intrathecal resiniferatoxin-induced analgesia. Journal of Pain. 12: 991-1003.3. Correia MAS, Mazumder K, Brás JLA, Firbank SJ, Zhu Y, Lewis RJ, York WS, Fontes CMGA andGilbert HJ (2011). The structure and function of an arabinoxylan-specific xylanase. Journal ofBiological Chemistry. 286: 22510-22520.4. Jena SN, Srivastava A, Singh UM, Roy S, Banerjee N, Rai KM, Singh SK, Kumar V, Chaudhary LB,Roy JK, Tuli R and Sawant SV (2011). Analysis of genetic diversity, population structure andlinkage disequilibrium in elite cotton (Gossypium L.) germplasm in India. Crop and PastureScience. 62: 859-875.5. Premkumar LS and Bishnoi M (2011). Disease-related changes in TRPV1expression and its implications for drug development. Curr Top Med Chem. 11(17): 2192-209.6. Raju M, Gupta V, Deeba F, Upadhyay SK, Pandey V, Singh PK and Tuli R (2011). A highly stableCu/Zn superoxide dismutase from Withania somnifera plant: gene cloning, expression andcharacterization of the recombinant protein. Biotechnology Letters. 33: 2057-2063.7. Sidhu OP, Annarao S, Chatterjee S, Tuli R, Roy R and Khaterpal CL ( 2011 ). Metabolic alterationsof Withania somnifera ( L.) Dunal fruits at different developmental stages by NMR spectroscopy.Phytochemical Analysis. DOI 10.1002 /pca.1307.8. Tiwari S, Mishra DK, Chandrashekar K, Singh PK and Tuli R (2011). Expression of d-endotoxinCry1EC from an inducible promoter confers insect protection in peanut (Arachis hypogaea L.)plants. Pest Management Science. 67: 137-145.9. Upadhyay SK, Sharad S, Singh R, Rai P, Dubey NK, Chandashekar K, Negi KS, Tuli R and SinghPK (2011). Purification and characterization of a lectin with high hemagglutination property isolatedfrom Allium altaicum. Protein Journal. 30: 374-83.58

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEEXTRAMURAL GRANTS AND FUNDINGS. No. Investigator Title of the Project Funded by1 Dr. Rakesh Tuli J. C. Bose Fellowship DST, Govt. ofIndia2 Dr. Sudhir P. Singh A novel strategy for developing scionplants of desired phenotype by using RNAidelivering rootstockSERB, DST,Govt. of India59

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEGOVERNANCENABI SOCIETYPresident: Minister, Science and TechnologyMember Secretary: Executive DirectorGoverning Body, NABIChairman: Secretary, DBTMember Secretary : Executive DirectorScientific Advisory Committee, NABIChairman: Eminent ScientistMember Secretary : Executive DirectorED/IMC, NABIPACAgri. BiotechnologyChair: DistinguishedScientistMember Secretary :Dean 1PACFood BiotechnologyChair: DistinguishedScientistMember Secretary :Dean 2PACNutritionalBiotechnologyChair: DistinguishedScientistMember Secretary :Dean 360

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEA. Members of NABI SocietySh. Vilasrao DeshmukhHon'ble Minister of Science & Technology andEarth Sciences,Ministry of Science & Technology and EarthSciences,Govt. of India, New Delhi(President)(July 12, 2011 to till date)Sh. Pawan Kumar BansalHon'ble Minister of Science & Technology andEarth Sciences,Ministry of Science & Technology and EarthSciences,Govt. of India, New Delhi(President)(Upto July 11, 2011)Dr. M. K. BhanSecretary,Department of Biotechnology,Ministry of Science & Technology,New DelhiMs. Anuradha MitraFinancial Advisor,Council of Scientific and Industrial Research,New DelhiDr. B. SesikeranDirector,National Institute of Nutrition,HyderabadDr. N. SathyamurthyDirector,Indian Institute of Science Education andResearch,MohaliDr. Akhilesh Kumar TyagiDirector,National Institute of Plant Genome Research,New DelhiDr. V. Prakash(Former Director, CFTRI)Distinguished Scientist,Council of Scintific and Industrial Research,MysoreDr. S. NagarajanFormer Chairperson,Protection of Plant Varieties and Farmers RightsAuthority,New DelhiDr. Rajesh KapurAdvisor,Department of Biotechnology,Ministry of Science & Technology,New DelhiDr. Rakesh TuliExecutive Director,National Agri-Food Biotechnology Institute,Mohali(Member Secretary)61

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEB. Governing Body (GB)Dr. M. K. BhanSecretary,Department of Biotechnology,Ministry of Science & Technology,New Delhi(Chairman)Dr. N. K. Ganguly(Former Director General- ICMR),Distuiguished Professor of Biotechnology,Translational Health Science & TechnologyInstitute,New DelhiMs. Anuradha MitraDr. V. PrakashFinancial Advisor,(Former Director, CFTRI)Council of Scientific and Industrial Research, Distinguished Scientist,New DelhiCouncil of Scintific and Industrial Research,MysoreDr. Manju SharmaPresident & Executive Director,Dr. Rajesh KapurIndian Institute of Advanced Research,Advisor, DBTGujaratCGO Complex,Lodi Road,Dr. C. R. BhatiaNew DelhiFormer Secretary,Department of Biotechnology,Dr. S. NagarajanNew DelhiFormer Chairperson,Dr. Ashok D. B. VaidyaProtection of Plant Varieties and Farmers RightsResearch Director,Authority,Kasturba Health Society Medical & ResearchNew DelhiCentre,Dr. B. Siva KumarMumbaiFormer Director,Dr. B. SesikeranNational Institute of Nutrition,Director,SecunderabadNational Institute of Nutrition,Dr. Joy K. RoyHyderabad Scientist D,Dr. N. SathyamurthyNational Agri-Food Biotechnology Institute,Director,MohaliIndian Institute of Science Education andDr. Sukhvinder P. SinghResearch,Scientist C,MohaliNational Agri-Food Biotechnology Institute,Dr. Akhilesh Kumar TyagiMohaliDirector,Sh. Shrikant Subhash MantriNational Institute of Plant Genome Research,Scientist C,New DelhiNational Agri-Food Biotechnology Institute,Dr. J. S. PaiMohali(Former Director, UICT)Dr. Rakesh TuliExecutive Director,Executive Director,Protein Foods & Nutrition DevelopmentNational Agri-Food Biotechnology Institute,Association of India,MohaliMumbai(Member Secretary)62

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEC. Finance CommitteeDr. M. K. BhanDr. Joy K. RoySecretary, Scientist D,Department of Biotechnology,National Agri-Food Biotechnology Institute,Ministry of Science & Technology,MohaliNew Delhi(Chairman)Dr. Sukhvinder P. SinghScientist C,Ms. Anuradha MitraNational Agri-Food Biotechnology Institute,Financial Advisor,MohaliCouncil of Scientific and Industrial Research,New DelhiSh. Shrikant Subhash MantriScientist C,Dr. Rakesh TuliNational Agri-Food Biotechnology Institute,Executive Director,MohaliNational Agri-Food Biotechnology Institute,MohaliSh. R. L. SharmaAssociate Director,Dr. Rajesh KapurNational Agri-Food Biotechnology Institute,Advisor,MohaliDepartment of Biotechnology,(Non-Member Secretary)Ministry of Science & Technology,New Delhi63

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTED. Scientific Advisory Committee (SAC)Dr. R. S. Paroda(Former DG, ICAR)Trust for Advancement of Agricultural Sciences,New Delhi(Chairman)Dr. C. R. BhatiaFormer Secretary,Department of Biotechnology,New DelhiDr. Deepak PentalVice-Chancellor,University of Delhi,New DelhiDr. B. Siva KumarFormer Director,National Institute of Nutrition,SecunderabadDr. V. Prakash(Former Director, CFTRI)Distinguished Scientist,Council of Scintific and Industrial Research,MysoreDr. Imran SiddiqiScientist,Centre for Cellular & Molecular Biology(CCMB),HyderabadDr. Anura V. KurpadProfessor of Physiology,St John's Medical College,BangaloreDr. H. P. S. SachdevSenior Consultant (Paediatrics),Sitaram Bhartia Institute of Science & Research,B-16, Kutub Institutional Area,New DelhiDr. Venkatesh RaoFormer Director,Central Food Technological Research Institute,MysoreDr. Arun SharmaOutstanding Scientist (Food Technology) ,Bhabha Atomic Research Centre,MumbaiDr. Rakesh TuliExecutive Director,National Agri-Food Biotechnology Institute,MohaliDr. Rajesh KapurAdvisor, DBTCGO Complex,Lodi Road,New DelhiDr. Akshay Kumar PradhanProfessor,Deptt. of Genetics,University of Delhi,South Campus, Dhaula Kuan,New Delhi64

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEE. Programe Advisory Committee (PAC): Agri-BiotechnologyDr. C. R. BhatiaFormer Secretary,Department of Biotechnology,New Delhi(Chairman)Dr. Kailash Chander BansalDirector,National Bureau of Plant Genetic Resources(NBPGR),New DelhiDr. Akhilesh Kumar TyagiDirector,National Institute of Plant Genome Research,New DelhiDr. G. K. GargDirector, ITRKrishidhan Research Foundation Pvt. Ltd.,Aurangabad Road,Jalna(Chairman)Dr. K. Madhavan NairDeputy Director,National Institute of Nutrition,HyderabadDr. (Prof.) Sunil Kumar MukharjeeScientist,International Centre for Genetic Engineering &Biotechnology,New DelhiDr. Kiran K. SharmaInternational Crops Research Institute forthe Semi-Arid Tropics (ICRISAT),HyderabadDr. Ramesh SontiDeputy Director,Centre for Cellular & Molecular Biology(CCMB),HyderabadDr. Rajesh KapurAdvisor, DBTCGO Complex,Lodi Road,New DelhiDr. Ashok K. SinghSenior Scientist & Programme Leader (Rice),Div. of Genetics,Indian Agricultural Research Institute,New DelhiDr. T. MohapatraPrincipal Scientist,NRC Plant Biotechnology,Indian Agricultural Research Institute,New DelhiDr. Rakesh TuliExecutive Director,National Agri-Food Biotechnology Institute,Mohali65

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEF. Programe Advisory Committee (PAC): Food and Nutrition BiotechnologyDr. V. Prakash(Former Director, CFTRI)Distinguished Scientist,Council of Scintific and Industrial Research,Mysore(Chairman-Food Biotechnology)Dr. B. Siva KumarFormer Director,National Institute of Nutrition,Secunderabad(Chairman-Nutrition Biotechnology)Dr. Appu RaoScientist,Central Food Technological Research Institute,MysoreDr. V. K. BatishEmeritus Scientist,Molecular Biology Unit,National Dairy Research Institute (NDRI),KarnalDr. K. Madhavan NairDeputy Director,National Institute of Nutrition,HyderabadDr. S. K. RoyEmeritus Professor & Consultant FAO,Indian Agricultural Research Institute (IARI),New DelhiDr. H. P. S. SachdevSitaram Bhartia Institute of Science & Research,B-16, Kutub Institutional Area,New DelhiDr. H. N. MishraProfessor,Agriculture & Food Engineering DepartmentIndian Institute of Technology,KharagpurDr. Bhupendar KhatkarChairman, Department of Food Technology,Guru Jambeshwar University of Science &Technology,HissarDr. M. C. VaradrajScientist,Central Food Technological Research Institute,MysoreDr. Rajesh KapurAdvisor, DBTCGO Complex,Lodi Road,New DelhiDr. Rakesh TuliExecutive Director,National Agri-Food Biotechnology Institute,Mohali66

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEH. Building CommitteeDr. G. P. TalwarDirector Research,Talwar Research Foundation,New DelhiDr. N. SathyamurthyDirector,Indian Institute of Science & EducationResearch,MohaliDr. P. K. SethChief Executive Officer,Biotech Park,LucknowSh. Sanjay GoelDirector (Finance),Department of Biotechnology,New DelhiDr. Rajesh KapurAdvisor,Department of Biotechnology,Ministry of Science & Technology,New DelhiEr. N. S. BhattiFormer Chief Engineer,Punjab Administration,ChandigarhDr. K. K. KaulFormer Chief Town Planner,Greater Mohali Area Development Authority,ChandigarhEr. S. K. JaitleyFormer Chief Engineer,UT Sectritariat,ChandigarhEr. S. L. KaushalFormer Chief Architect Punjab,ChandigarhMs. Balwinder SainiChief Architect,Govt. of Punjab,ChandigarhEr. Kuldeep SinghFormer Chief EngineerChandigarh Adminstration,ChandigarhDr. S. B. KattiScientist G,Central Drug Research Institute,LucknowDr. U. V. PathreScientist,National Botanical Research Institute,LucknowDr. Jagdeep SinghCoordinator,Indian Institute of Science Education andResearch,MohaliEr. A. A. MalikSupretendent Engineer,National Botanical Research Institute,LucknowDr. Rakesh TuliExecutive Director,National Agri-Food Biotechnology Institute,Mohali(Chairman)67

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEHUMAN RESOURCE69

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEI. Research Facultyi. Regular:S. No123456789101112Name Designation Date of JoiningDr. Rakesh Tuli Executive Director 08-02-2010Dr. Vikas Rishi Scientist E 01-03-2012Dr. Joy K. Roy Scientist D 09-08-2010Dr. Ajay K. Pandey Scientist D 14-11-2011Dr. Siddharth Tiwari Scientist C 28-07-2010Sh. Shrikant Subhash Mantri Scientist C 18-08-2010Dr. (Ms.) Monika Garg Scientist C 30-11-2010Dr. Sukhvinder P. Singh Scientist C 06-12-2010Dr. Kanthi K. Kiran Scientist C 02-09-2011Dr. Mahendra Bishnoi Scientist C 16-12-2011Dr. Koushik Mazumder Scientist C 01-02-2012Dr. Nitin K. Singhal Scientist C 02-03-2012ii. On Contract:S. No Name Designation Date ofJoiningII. Technical and Engineering SupportS. No Name Designation Date of Joining1 Sh. E. Subramanian Computer Operator 27-02-2010234512 Dr. Nishima Project Scientist 04-01-20123Dr. Shailesh Sharma Project Scientist 02-01-2012Dr. Sudhir P. Singh Project Scientist 16-01-20124 Ms. Vandana Mishra Project Scientist 23-01-2012Er. Jatin Singla Assistant Engineer (Civil) 21-01-2011Ms. Aakriti Gupta Senior Tech Assistant 22-02-2011Sh. Jagdeep Singh Senior Tech Assistant 01-03-2011Sh. Sukhjinder Singh Computer Operator 23-02-20126 Sh. Jaspreet Singh Assistant Engineer (Civil) 19-03-2012III. AdministrationS. No Name Designation Date of Joining1 Sh. Rattan Lal Sharma Associate Director(Accounts and Finance)2345625-5-2011Sh. S. Krishnan Store & Purchase Officer 10-03-2010Sh. Vikram Singh Administrative Officer 01-04-2011Sh. Suneet Verma Finance Officer 15-09-2011Sh. Sabir AliMs. Hema RawatExecutive Assistant(Administration)Executive Assistance(Accounts)7 Sh. Vishal Kumar Management Assistant(Accounts)21-01-201101-04-201108-09-201171

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEIV. Human Resource Developmenti. Research Scholars:S.No Name Designation Date of Joining1 Sh. Jitendra Kumar Senior Research Fellow 04-06-20102 Sh. Yogesh Gupta Junior Research Fellow 09-08-20103 Ms. Anuradha Singh Junior Research Fellow 11-09-20104 Sh. Rohit Kumar Junior Research Fellow 06-06-20115 Ms. Anita Kumari Junior Research Fellow 29-08-20116 Sh. Anshu Alok Junior Research Fellow 09-09-20117 Ms. Shaweta Arora Junior Research Fellow 09-09-20118 Ms. Manpreet Kaur Saini Junior Research Fellow 09-09-20119 Sh. Jitesh Kumar Junior Research Fellow 09-09-201110 Sh. Gaurab Das Junior Research Fellow 10-10-201111 Sh. Kaushal Kumar Bhati Junior Research Fellow 14-11-201112 Ms. Anuradha Swami Junior Research Fellow 23-02-201213 Ms. Roohi Bansal Junior Research Fellow 24-02-201214 Ms. Monica Sharma Junior Research Fellow 01-03-201215 Sh. Raja Jeet Junior Research Fellow 12-03-2012ii. Trainees:S.No Name Designation Date of Joining1 Sh. Vikas Bindal Trainee 02-01-20122 Ms. Arushi Sharma Trainee 02-01-20123 Ms. Natasha Paul Trainee 02-01-20124 Ms. Manila KashyapTrainee02-01-20125 Sh. Ravindra Dhakad Trainee 02-01-20126 Sh. Anuj Kumar Pandey Trainee 03-01-201278910Sh. Rakesh Kumar Trainee 03-01-2012Ms. Iram Preet Kaur Trainee 04-01-2012Ms. Arti Katoch Trainee 04-01-2012Ms. Guneet Kaur Trainee 04-01-201211 Ms. Lipika Sahota Trainee 09-01-20121213Ms. Ramandeep Kaur Trainee 09-01-201214 Sh. Nishant Garg Trainee 03-02-201215Sh. Deepak Soni Trainee 13-01-2012Sh. Shashank Maheshwari Trainee 03-02-201272

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEPROGRESS OF INFRASTRUCTUREAT MAIN CAMPUS73

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEProposed Master Plan of the main campus,Sector-81, Ajitgarh (Mohali)75

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEMain entrance gate of the campus at sector-81, Mohali.Low lying area at the main campus and a view of IISER campus on the other side of boundary wall.76

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEPROGRESS OF INFRASTRUCTUREAT INTERIM FACILITY77

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEDevelopment of NABI interim facilitystthInauguration of construction work at 1 floor at interim facility: July 4 , 2011stUpper and lower left: Analytical instruments rooms at 1 floor.stUpper Right: New laboratories at 1 floor.stLower right: Plant tissue culture and animal cell culture facility developed at 1 floor.79

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEstPlant Tissue Culture facility at 1 floor.stScientists working at Animal Cell Culture facility, 1 floor.80

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEIMPORTANT EVENTS81

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEImportant events:promote Indo-Canada partnershipthdevelopment activities: August 14 , 2011.1. Independence Day celebration: Augustth15 , 2011. Dr. Rakesh Tuli, hoisted the 6. Visit of Nutrition Science andNational flag at interim facility.Biotechnology Faculty SelectionthCommittee experts: October 29 , 2011.2. Tree plantation programe at the mainthcampus: August 18 2011. Dr. Rakesh Tuli 7. Visit of Food Science and Biotechnologyand staff participated in the tree plantation Faculty Selection Committee experts :th thprograme. Pollution tolerant species were November 27 – 28 , 2011.planted along with the boundary wall ofNABI campus with the objective ofmaking the campus green and colourful.8. Visit of National Dairy Research Institutethdelegates: December 27 , 2011.th3. Republic Day celebration: January 26 , 9. Second Scientific Advisory Committee2012. Dr. Rakesh Tuli, hoisted the national(SAC) and Programe Advisory Committeeth thflag at interim facility.(PAC) meeting : February 18 - 19 , 2012.nd4. The 2 Foundation Day of NABI was10. Visit of University of Agriculturalthcelebrated at interim facility: FeburarySciences delegates: February 27 , 2012.th18 , 2012. Padma Bhushnan Dr. R. S.Paroda, former DG, ICAR & Secretary,National/International visits of NABI staff:Department of Argicultural Research and 1. Dr. Joy K. Roy was invited to participate inEducation (DARE) delivered thethe symposium organized by the IndianFoundation Day lecture “HarnessingNational Science Academy (INSA) underbiotechnology in Indian agriculture:its Bilateral Exchange Programe with theChallenges and Opportunities”. PadmaGerman National Academy of Sciences,Bushna Dr. N K Ganguly, former DG,Leopoldina on “Plant Biology” which wasICMR gave Presidential Address on “Foodth thheld during October 18 -20 , 2011 at NewSecurity and Nutritional Challenges forDelhi. He also presented his research workMarginalised Populations”. They alsoentitled “Towards understanding ofreleased the Foundation Year Annualmolecular basis of processing quality inReport 2010-2011 of the institute.wheat”.Meetings2. Dr. Sudhir P. Singh visited, Canadian LightSource, Saskatoon, Canada from1. Visit of Agri-Biotechnology Faculty th ndthSeptember 26 to October 2 , 2011 toSelection Committee experts : July 12 ,perform experiments on mineral2011.localization in the seed tissues of wheat.2. First Programe Advisory CommitteeThe Beam time was awarded by theth(PAC) meeting : July 26 , 2011.Canadian Light Source to perform X-RayFluorescence and XANES experiments on3. First Scientific Advisory Committee VESPERS beam line.nd(SAC) meeting : August 2 , 2011.3. Sh. Shrikant Mantri, attended a seminar on4. Third Governing Body (GB) meeting :Genomeet (A Decade of Genomics) held atndAugust 2 , 2011.stIGIB, New Delhi on January 1 , 2012.5. Visit of Canadian Delegates fromSaskatoon-cluster, Saskatchewan, to4. Dr. Sudhir P. Singh attended the first ICCInternational Grains Conference held at83

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEth thNew Delhi during January 16 -18 , 2012. International visitors to NABIHe presented his work entitled “New1. Canadian delegates visited NABI campusinsights into iron transport from maternaland interacted with NABI Scientists:tissues to endosperm in mature wheat seedthAugust 14 , 2011.using synchrotron radiation”.2. Dr. Bikram Singh Gill from Kansas State5. Dr. Joy K. Roy attended the first “ICC University, Kansas, USA visited NABI onInternational Grains Conference”, whichrdJanuary 23 , 2012. He discussed researchth thwas held during January 16 - 18 , 2012 atprogress with NABI scientists. He gave hisNew Delhi. He presented his work entitledpresentation on recent approaches in wheat“Gene discovery for improvement ofgenomics.processing quality in bread wheat”.3. Dr. Harbans Bariana from University of6. Dr. Nishima, participated in a seminar on Sydney, Sydney, Australia, Plant Breeding“Green Chemistry” held at Ludhiana onInstitute, Australia visited NABI onththFebruary 24 , 2012.Febuary 9 , 2012 to share his thoughts andinteracted with NABI faculty in the area of7. Dr. Rakesh Tuli, attended Biotechnology- plant breeding and genetic resources.Industry Research Assistance Councilth thmeeting at Australia during March 6 -10 , 4. Prof. Leon A. Terry from Cranfield2012 and visited to Queensland UniversityUniversity, Bedfordshire, England visitedthTechnology, Brisbane, Australia to discussNABI on Febuary 15 , 2012 and discussedabout the project on banana improvement.about his research work.84

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEPHOTO-GALLERY85

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEAgri-Biotechnology Faculty Selection Committee:thJuly 12 , 2011Visit of experts to interim facility. Dr. Joy K. Roy explaining his work on Affymetrix microarray to(from left) Dr. A. K. Tyagi, Dr. C. R. Bhatia, Dr. J. P. Khurana, Dr. P. K. Gupta, Dr. Imran Siddiqiand Dr. Rakesh Tuli.Visit of experts to the main campus, Sector-81, Mohali.87

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEthFirst PAC Meeting: July 26 , 2011PAC meeting being chaired by Dr.V. Prakash.Distinguished experts from Agri, Food and Nutrition Biotechnology during the PAC meeting.88

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEndFirst SAC Meeting: August 2 , 2011First SAC meeting being addressed by Dr. M.K. Bhan and chaired by Dr. C.R. Bhatia.Dr. Rakesh Tuli, presenting the research plans of NABI to PAC and SAC members.89

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEndThird Governing Body Meeting: August 2 , 2011The third GB meeting being chaired by Dr. M. K. Bhan, Secretary, DBT,Dr. (Mrs.) Manju Sharma, Former Secretary, DBT and member GB is seen on the right.Dr. Rakesh Tuli, discussing the progress, oppurtunities and challanges in Agri-Food researches withthe distinguished GB members.90

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEIndependence Day Celebrationsthat NABI: August 15 , 2011Dr. Rakesh Tuli, hoisted the national flag at NABI interim facility and addressed the staff.Independence Day celebrations at the main campus, Sector 81, Mohali.91

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEthTree Plantation Programe at the Main Campus: August 18 , 2011Tree plantation programe was organised at the main campus, Sector 81, Mohali. Dr. Rakesh Tuliand staff participated in the programe. Pollution tolerant species were planted along the boundarywall to make campus look green and colourful, under the guidance of Dr. S. C. Sharma, former Dy.Director, NBRI, Lucknow.92

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEVisits of DelegatesVisit of Canadian delegates from Saskatoon-cluster, Saskatchewan, to promote Indo-Canadathpartnership: August 14 , 2011. From Left: Sh. G. V. Shankar, Dr. Venkatesh Meda, Dr. Rakesh Tuli,Dr. Sudhir P. Singh and Dr. Robert (Bob) Tyler.thVisit of Scientific delegation from National Dairy Research Institute: December 27 , 2011.93

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEthRepublic Day Celebrations at NABI: January 26 , 2012Dr. Rakesh Tuli, hoisted the national flag.Republic day celebrations at NABI interim facility.94

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTESecond PAC Meeting: February 18th, 2012First row from left: PAC meetings being chaired by Dr. V. Prakash (PAC – Food Biotechnology) ,Dr. B. Siva Kumar (PAC – Nutrition Biotechnology) and Dr. R. S. Paroda (PAC – AgriculturalBiotechnology).Second row from left: Discussion during the combined PAC meeting of Agri, Food and NutritionBiotechnology.Third row: Visit of the experts to the main campus at Sector 81, Mohali.95

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEthSecond SAC Meeting: February 19 , 2012SAC meeting being chaired by Dr. R.S. Paroda, Chairman SAC and former Director General, ICAR.Experts in Agri, Food and Nutrition Biotechnology having discussion with faculty, duringthe SAC meeting.96

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTESecond Foundation Day: February 18th, 2012First row from left: Dr. R.S. Paroda, Dr. N.K. Ganguly, Dr. B. Siva Kumar and other distinguishedguests invited on the occasion. Dr. R.S. Paroda was the Chief Guest and Dr. N.K. Ganguly presidedover the function.Second row from left: Dr. Rakesh Tuli, reading Annual Report of NABI. Signing of MoU betweenNIPER and NABI.Third row from left: Dr. R. S. Paroda delivering a talk on “Harnessing Biotechnology in IndianAgriculture: Challenges and Opportunities”. Dr N.K. Ganguly gave Presidential address “FoodSecurity and Nutritional Challenges for Marginalised Populations” on the occasion. Dr. Ajay K.Pandey giving the vote of thanks.97

NATIONAL AGRI-FOOD BIOTECHNOLOGY INSTITUTEAnnual Accounts for the year 2011-12:lllThe financial resource of the institute is thecore grant provided by the Department ofBiotechnology, Govt. of India under nonrecurring& recurring components.The institute received the core grant of Rs.2,358.00 lac in the year 2011-12.Annual accounts for the year 2011-12 areFINANCIALSlprepared by the institute on the basis ofaccrual system of accounting usingstandard format of accounts prescribed bythe Government of India for CentralAutonomous Bodies.M/s Raj Gupta & Co., (CharteredAccountant) Chandigarh, the statutoryauditors of the institute have audited theaccounts.Financial Position:-S.No.(Figure in Rupees)Particulars As on 31-03-2011 As on 31-03-2012A. Capital Fund and Liabilities1. Capital Fund 22,52,20,686 36,08,87,7052. Current Liability and Provisions 11,16,888 69,72,351B. AssetsTotal 22,63,37,574 36,78,60,0561. Fixed Assets 11,84,15,380 18,07,26,3632. Capital Work-in-Progress 76,20,000 3,69,11,6303. Current Assets ,Loans & Advances etc. 10,03,02,194 15,02,22,063Total 22,63,37,574 36,78,60,056C. Receipts of the Institute1. Grants from DBT 22,22,24,014 23,58,00,0002. Internal Receipts /Income 23,01,994 91,79,908Total 22,45,26,008 24,49,79,908D. Utilization1. Equipment 12,08,76,581 7,42,44,8352. Furniture and other Fixed Assets 44,67,606 62,47,8153. Manpower 77,35,259 2,12,64,2574. R&D Expenses 1,07,13,216 1,54,34,5105. Administrative Expenses 2,89,08,896 5,44,32,454E. Payables/Outstandings 11,16,888 69,72,351F. Advances / Receivables 1,63,49,345 1,65,25,554G. Margin Money for LCs 5,50,00,000 8,27,47,849H. Closing Bank Balance (Excluding MarginMoney )2,89,52,849 5,09,48,66098

C-127, Industrial Area, Phase 8, Ajitgarh (Mohali), Punjab, India - 160071