INTRODUCTION OF GLYOXALASE I GENE FOR SALT TOLERANCE INTO RICE (Oryza sativa L.) VARIETIES ‘PR118’ AND ‘KITAAKE’ THROUGH PARTICLE BOMBARDMENT

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dc.contributor.advisorAjinder Kaur
dc.contributor.authorSaroj Kumar Sah
dc.date.accessioned2017-07-10T04:51:13Z
dc.date.available2017-07-10T04:51:13Z
dc.date.issued2013
dc.description.abstractThe present investigation dealing with introduction of glyoxalase I gene for salt tolerance into rice (Oryza sativa L.) through particle bombardment was carried out using two varieties PR118 (indica) and Kitaake (japonica). Calli were induced from both the varieties on MS medium supplemented with 2,4-D (3.0 mgL-1) + BAP (0.25 mgL-1) + proline (600 mgL-1) + maltose (40 gL-1) + phytagel (3 gL-1). Embryogenic calli were subcultured on shoot regeneration MS medium supplemented with BAP (4.0 mgL-1) + NAA (0.2 mgL-1) + sucrose (30 gL-1) + phytagel (2 gL-1) + agar (8 gL-1). An attempt was made to introduce GlyI gene into both the varieties. Mature seed-derived embryogenic calli were used as explants for all the transformation experiments. Success was achieved in both the varieties. Using GlyI gene, in PR118, out of 2600 calli bombarded, 249 putative transgenic plants were regenerated on medium containing hygromycin (30 mgL-1). Likewise, in Kitaake, out of 615 calli bombarded, 469 putative transgenic plants were regenerated on medium containing hygromycin (30 mgL-1). Among the 249 regenerants of PR118, 13 plants were PCR positive and in case of Kitaake, among 615 plants 41 showed PCR positive results. Further, these plants were grown to maturity in transgenic glass house. Presence of copy number of transgene was done by Real Time PCR. In total, a set of 10 PCR positive plants (5 of PR118 and 5 of Kitaake) 10 samples were analyzed, among them sample number four has single copy gene and rest has multiple copy ranging from 2-14 copies. To conclude tissue culture base line has been established in two varieties PR118 and Kitaake. Using this baseline, a total of 54 PCR positive transgenic plants were developed. The protocol developed here is genotype independent and is suitable for japonica as well as indica varieties.en_US
dc.identifier.urihttp://krishikosh.egranth.ac.in/handle/1/5810024740
dc.keywordsKeywords: Callus induction, Regeneration, MS medium, Transformationen_US
dc.language.isoenen_US
dc.publisherPAU Ludhianaen_US
dc.subBiotechnology
dc.subjectnullen_US
dc.themeRICE (Oryza sativa L.)en_US
dc.these.typeM.Scen_US
dc.titleINTRODUCTION OF GLYOXALASE I GENE FOR SALT TOLERANCE INTO RICE (Oryza sativa L.) VARIETIES ‘PR118’ AND ‘KITAAKE’ THROUGH PARTICLE BOMBARDMENTen_US
dc.typeThesisen_US
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