几丁质酶-葡聚糖酶双价基因导入滇型杂交稻恢复系提高稻瘟病抗性的研究

A Study on Introduction of Chitinase Gene and β-1,3-Glucanase Gene into Restorer Line of Dian-type Hybrid Rice (Oryza sativa L.) and Enhanced Resistance to Blast (Magnaporthe grisea)

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。

Plasmid pBLGC containing chitinase gene from Phaseolus limensis and β 1,3 glucanase gene from Nicotiana tabacum was bombarded into the restorer line 'Nan29' of Dian type hybrid rice ( Oryza sativa L.ssp. japonica ) from Yunnan province of South west China.93 regenerants were obtained from the calli that were resistant to G418 (100 to 150 mg/L) on NB medium.Using β 1,3 glucanase gene as the probe,17 of the regenerants were identified to be transgenic lines by dot blotting and the foreign genes construction were intergrated into the genomes of T 1 lines by Southern blotting hybridization.Two foreign genes were inherited stably to T 4 generation according to PCR results of the lines.The resistance to rice blast of six transgenic linges were evaluated by inoculating four violent biological races of Magnaporthe grisea from Yunnan province and inducing the disease in the field.The results indicated that the resistance to rice blast of transgenic lines were enhanced to varying degrees compared with the receptor line and the transgenic lines could be used in rice blast resistant breeding.