转cry1Ab13基因玉米新种质的创制

发布时间：2018-06-11 22:57

  本文选题：抗虫性 + cryAb基因　； 参考：《华南农业大学学报》2016年05期

【摘要】：【目的】构建包含抗鳞翅目害虫基因cry1Ab13的重组植物表达载体,并利用其创制对玉米螟Ostrinia furnacalis具有优良抗性的转基因玉米Zea may L.新种质。【方法】利用同源重组法将cry1Ab13基因连接到表达载体pCAMBIA3300-Bar上,获得以抗除草剂Bar基因为筛选标记的植物表达载体pCAMBIA3300-cry1Ab13-Bar。通过农杆菌介导法转化玉米自交系H99幼胚,对再生植株进行逐代除草剂筛选、PCR检测及T2代植株的Southern blotting检测、实时荧光定量PCR检测,并对转基因植株进行田间及室内抗虫性鉴定。【结果】构建了cry1Ab13基因的植物表达载体,转化玉米获得3株高抗玉米螟和1株抗玉米螟的T2代转基因植株。Southern blotting证明cry1Ab13基因已经整合到玉米基因组中,实时荧光定量PCR结果表明cry1Ab13基因已经在玉米植株内表达。抗虫性鉴定结果表明,与对照相比转基因植株对玉米螟的抗性显著提高。【结论】将cry1Ab13基因导入玉米并成功表达,显著提高了转基因玉米对玉米螟的抗性,为抗虫玉米新种质的创制奠定了基础。
[Abstract]:[objective] to construct a recombinant plant expression vector containing the Lepidoptera pest resistance gene cry1Ab13, and to create a transgenic maize may L- Zea may with excellent resistance to Ostrinia furnacalis. [methods] the cry1Ab13 gene was ligated to the expression vector pCAMBIA3300-Bar by homologous recombination method, and the plant expression vector pCAMBIA3300-cry1Ab13-Barwas obtained by screening marker pCAMBIA3300-Bar. The immature embryos of maize inbred line H99 were transformed by Agrobacterium tumefaciens. The regenerated plants were screened by herbicides and detected by Southern blotting and real-time fluorescence quantitative PCR. The plant expression vector of cry1Ab13 gene was constructed. Three transgenic plants with high resistance to corn borer and one plant resistant to corn borer were obtained. Southern blotting showed that the cry1Ab13 gene had been integrated into the maize genome. The results of real-time fluorescence quantitative PCR showed that the cry1Ab13 gene had been expressed in maize plants. The results of insect-resistance identification showed that the resistance of transgenic plants to corn borer was significantly higher than that of control. [conclusion] cry1Ab13 gene was introduced into maize and expressed successfully, which significantly improved the resistance of transgenic maize to corn borer. It lays a foundation for the creation of new insect-resistant maize germplasm.
【作者单位】： 吉林农业大学生命科学学院;
【基金】：吉林省省级粮食生产发展专项(2015001) 吉林农业大学科研启动基金(201242)
【分类号】：S513
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本文编号：2007005