cry2Ah-M基因杀虫活性研究

发布时间：2018-05-20 13:25

  本文选题：棉铃虫 + cryAh-M　； 参考：《中国农业科技导报》2017年04期

【摘要】：转Bt基因抗虫棉经过20多年的研究和生产应用,虽然取得了巨大的生态和经济效益,但棉铃虫的抗性风险也逐渐增加,因此,通过发掘不同杀虫机理的Bt基因,不仅可以提高杀虫效率,而且有效的避免或延迟昆虫的抗性。依据棉花遗传密码子偏好性,对来源于苏云金芽胞杆菌(Bacillus thuringiensis,Bt)新型Bt基因cry2Ah-like进行了密码子优化改造,利用农杆菌介导法导入烟草。通过荧光定量PCR方法,Southern blot和Western blot方法,证明了转基因烟草中外源基因在RNA和蛋白水平上均稳定表达,并且已整合到烟草基因组中。生物活性检测结果表明,饲喂转基因烟草叶片4 d后,棉铃虫幼虫校正死亡率达86.59%,达到高抗水平,进一步证明密码子优化后的cry2Ah-M基因能够高效表达且具有较高的杀虫活性,是有效防治棉铃虫的新型基因。
[Abstract]:Transgenic BT cotton has been studied and applied for more than 20 years. Although it has achieved great ecological and economic benefits, the resistance risk of Helicoverpa armigera has also gradually increased. Therefore, the BT gene of different insecticidal mechanism was explored. It can not only improve insecticidal efficiency, but also effectively avoid or delay insect resistance. Based on the preference of cotton genetic codon, a new BT gene cry2Ah-like from Bacillus thuringiensis Btwas optimized and introduced into tobacco by Agrobacterium tumefaciens. Southern blot and Western blot methods were used to confirm the stable expression of transgenic tobacco genes at the RNA and protein levels, and they were integrated into the tobacco genome. The results of bioactivity test showed that the corrected mortality of cotton bollworm larvae reached 86.59 and reached a high level of resistance after feeding transgenic tobacco leaves for 4 days. It was further proved that the codon optimized cry2Ah-M gene could be expressed efficiently and had higher insecticidal activity. It is a new gene to control cotton bollworm effectively.
【作者单位】： 新疆农业大学农学院;中国农业科学院生物技术研究所;中国农业科学院植物保护研究所;
【基金】：国家转基因生物新品种培育重大专项(2014ZX08005-004)资助
【分类号】：S435.62
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本文编号：1914789