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2013 - 20142
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Subject: Agricultural Biotechnology × End date: 2014 × Start date: 2010 × Type: Thesis × Thesis Type: M.Sc ×
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    DEVELOPMENT OF LINKAGE MAP IN SYNTHETIC43 X WH542 RIL POPULATION AND MAPPING OF POWDERY MILDEW RESISTANCE GENES
    (Punjab Agricultural University, Ludhiana, 2014) Meha Sharma
    Powdery mildew, caused by Blumeria graminis f.sp. tritici is a major foliar disease of wheat worldwide.Present investigation was undertaken for generation of linkage map in Synthetic43 X WH542 RIL population and mapping of powdery mildew resistance gene using this linkage map. A synthetic wheat, Synthetic 43, derived from CIMMYT is found to be resistant to rust and powdery mildew(PM) pathogens, besides having several agronomically important characters. A RIL population was generated by crossing Synthetic 43 with cultivated wheat variety WH542 through single seed descent method. Inheritance studies in RIL population revealed monogenic inheritance of powdery mildew resistance both at seedling and adult plant stage. This gene was temporarily designated as PmT. For generating linkage map maximum of 80 polymorphic SSR markers have been selected from D genome while 34 were selected from A and B genome corresponding to low polymorphic regions of D genome.These 114 SSR subjected to linkage analysis using MAPDISTO (1.7.6.5) software at LOD 2.5 and only 52 markers were mapped generating a linkage map of length 693.5cM with 13 linkage groups. These linkage groups were aligned with already known consensus map of wheat and 7 homeologous groups were generated. The size of these seven homeologous groups varied from 54.4cM (homeologous group 2) to 153.9cM (homeologous group 5). The marker order was generally collinear with the already published bread wheat consensus map with only 12 marker showing discrepancies in order and five SSRs mapped onto a different homeologous group. Out of 52 mapped SSRs 11 showed segregation distortion from expected ratio of 1:1.Using this linkage map, PmT gene was mapped on homeologous group 7 at a distance of 11.6cM distal to SSR Xwmc150. Xwmc150 has been mapped on the long arm on of chromosome 7D according to wheat consensus map
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    ESTABLISHING EFFICIENT IN VITRO REGENERATION PROTOCOL AND GENETIC TRANSFORMATION OF PIGEONPEA (Cajanus cajan L. Millsp.) THROUGH BIOLISTIC GUN USING cry1Ac GENE
    (Punjab Agricultural University, Ludhiana, 2013) Atul Dev; Ajinder Kaur
    The present investigation entitled “Establishing efficient in vitro regeneration protocol and genetic transformation of pigeonpea (Cajanus cajan L. Millsp.) through biolistic gun using cry1Ac gene” was carried using two varieties of pigeonpea ICPL87 and ICPL 88039. Regeneration from callus and direct organogenesis, both approaches were used to establish a tissue culture baseline to carry out transformation. Different explants viz., leaves, cotyledons, epicotyls, embryonic axes, mature and immature embryos were used for callus induction. None of the calli regenerated into plants on any of the growth hormone combinations and concentrations used. Organogenesis approach worked well with the induction of multiple shoots from cotyledonary nodes. Both the varieties exhibited a high frequency of shoot buds (60-65%) on MS + 5mgL-1 BAP medium. Elongation of the shoot buds was carried out on MS + 30 mgL-1 adenine sulphate medium and an average of 5-6 shoots per explant was obtained. Rooting and hardening were done ex vitro with a success rate of 75-94%, using a rooting mixture ROOTEXTM. For genetic transformation, biolistic approach was used to introduce cry1Ac gene. Mature seeds after 12 days of culturing produced axillary meristems in cotyledonary nodes that were used as target tissue for bombardments. Bombarded explants were kept on shoot induction medium for further induction of shoot buds for 10-12 days that were later transferred to shoot elongation medium. In total, 8 transformation experiments were conducted and a total of 326 axillary meristem explants (169 of ICPL 87 and 157 of ICPL 88039) were bombarded, from which 416 shoots in ICPL 87 and 384 shoots in ICPL 88039 were regenerated. Transformation results were promising in both the varieties with transformation efficiencies of 1.18% in ICPL 87 and 3.18% in ICPL 88039