Annual Report 2011-2012 - Nabi.res.in

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