【摘要】 将抗病虫基因与广亲和基因聚合在一起可为两系杂交稻更广泛的应用奠定基础。本研究将去选择标记的包含2个抗虫基因(sbk和sck)的质粒载体(pCDMARUBA-Hyg)采用根癌农杆菌介导法转入兼抗水稻条纹叶枯病的广亲和水稻品种Dular中。通过常规PCR检测,获得18份不具有选择标记的转基因阳性植株,Southern杂交分析结果表明其中15份为单拷贝插入,对这15份单拷贝转基因植株进行RT-PCR分析,结果抗虫基因都能正常表达。人工接虫鉴定结果表明,转入抗虫基因显著提高了水稻品种Dular抗螟虫的能力;同时利用Dular本身的广亲和基因(S5n)标记和与抗条纹叶枯病基因(qSTV-11b和qSTV-11c)的连锁标记进行分子检测,其结果表明这15份单拷贝转基因植株都具有广亲和与抗条纹基因的特征带;与不同籼稻和粳稻配组杂种F1的结实率和条纹叶枯病在田间自然发病状况下的调查结果表明,这些单拷贝植株不但具有广亲和与抗水稻条纹叶枯病的能力,而且其抗螟虫能力显著提高。更多还原

【Abstract】 It can lay a solid foundation for the wider application of two-line hybrid rice by pyramiding disease resistance genes,insect resistance genes and wide compatible genes.In this study,two insecticidal marker-free gene sbk and sck in the plasmid of pCDMARUBA-Hyg have been transferred into the rice varieties Dular,which itself has the S5n gene and three stripe virus resistant gene loci mediated by Agrobacterium tumerfaciens.By conventional PCR,18 marker-free transgenic plants were gained,and Southern blot analysis showed that 15 of them were single copy insertion,and these 15 single-copy transgenic plants could normally express the insect-resistant gene by RT-PCR analysis.Results of feeding rice stem borer by manual also indicated that the ability of transgenic plants was greatly increased.While takingadvantage oftheir wide compatible gene(s5n) marker and with anti-stripe gene(qSTV-11b and qSTV-11c) of the linkage markers for molecular detection,the results indicated that these 15 single-copy transgenic plants had wide compatiblities and the characteristics of resistance to stripe belt.Field test results about the setting percentage of the hybirds between these single-copy transgenic plants and some indica/japonica rice varieties,and the cases of rice stripe virus showed that these single-copytransgenic plants not onlyretained themselves internal wide compatible and stripe virus resistant property,resistant ability of rice stalk borer had also been significantly increased.更多还原