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将含有菜豆 (Phaseoluslimensis)几丁质酶基因和烟草 (Nicotianatabacum) β 1,3 葡聚糖酶基因的pBLGC(16 5kb)质粒用基因枪法导入滇型杂交稻 (OryzasativaL .ssp .japonica)恢复系“南 2 9”中 ,总计获得 93个转化再生植株 ,以β 1,3 葡聚糖酶基因制备探针对T1 代株系进行Southern杂交分析 ,17个株系为杂交阳性 ,证实外源基因完整结构已整合到水稻基因组中 ;连续多代的PCR追踪检测证实外源基因已遗传至T4 代 ;对所获得的 6个PCR检测阳性T4代品系进行了稻瘟病菌 (Magnaporthegrisea)生理小种接种鉴定和稻瘟病大田诱发鉴定 ,结果表明 ,转基因品系对稻瘟病的抗性较受体对照大幅度提高 ,获得了稻瘟病新抗源 ,但不同品系稻瘟病抗性并不相同。
The pBLGC (16 5kb) plasmid containing Phaseoluslimensis chitinase gene and Nicotianatabacum β 1,3-glucanase gene was introduced into Oryzasativa L. ssp. Japonica restorer line by gene gun method. Southern 2 9 ", a total of 93 transformed and regenerated plants were obtained. Southern hybridization analysis was carried out on the T1 generation lines by using β 1,3 glucanase gene preparation probes, and 17 lines were positive for hybridization and confirmed that the foreign gene was intact The structure was integrated into the genome of rice. PCR tracing for successive generations confirmed that the foreign gene has been inherited to T4 generation. The obtained 6 PCR-positive T4 generation lines were identified for inoculation with Magnaporthe grisea And rice blast induced identification. The results showed that the resistance of the transgenic lines to rice blast increased greatly compared with that of the recipient, and new resistant sources of blast were obtained. However, the blast resistance of different lines was not the same.