转修饰cry1Ac基因籼稻明恢81经花药培养获得抗虫DH系

对基因枪法获得的明恢81转修饰的cry1Ac基因当代植株进行花药培养，共接种花药2600枚，获得83份花培植株，其中双倍体植株43份，单倍体植株40份，PCR结果表明含有cry1Ac基因的植株55份，花培植株群体中转基因与非转基因植株的比值为2：1（55／28），进一步结合Southern blot和ELISA分析，于花培植株当代筛选到转基因纯合株系36份，外源蛋白表达量上，花药来源于同一克隆的DH系的不同植株之间基本一致，最高的Cry1Ac含量达0．25％，田间抗虫性试验表明，经花药培养纯合获得的部分转基因纯合系植株对二化螟（chilo suppressalis)表现出高抗，而且主要农艺性状保持不变，以上结果表明水稻花药培养可以加速转基因的纯合与良种利用。

Anther Culture Generated Stem Borer-resistance DH Lines of Minghui 81( Oryza sativa L. subsp . indica ) Expressing Modified cry 1Ac Gene

2600Anthers from T 0 modified cry1Ac -transgenic rice lines of Minghui 81, an elite restoring line of commercial CMS indica hybrid rice, were cultured on SK3 media. 83 green plantlets were recovered, 43 double haploid (DH) and 40 haploid among them. Results of PCR analyzes indicated that 55 plants of 83 were harbored the cry1Ac gene, and the ratio of cry1Ac -positive against cry1Ac -negative was 2:1 (55/28). 36 putative transgenic DH plants were further confirmed by Southern blot. ELISA detection showed that Cry1Ac level in different transgenic rice plants of the same cry1Ac -DH clone was almost equal and the highest one amount to 0.25% of the total soluble protein. Pest insect-resistant bioassay at field trials demonstrated that some of the homozygous cry1Ac -transgenic rice plants not only showed high-level resistance against striped stem borer (Chilo suppressalis) but also retained elite agronomy characters. These results demonstrated that rice anther culture has a great value in rice molecular breeding.