亚洲玉米螟对Cry9Ee与Cry1Ab蛋白的交互抗性分析

doi:10.16409/j.cnki.2095-039x.2015.06.010

摘要/Abstract

摘要： Cry9Ee是近年来发现的对亚洲玉米螟Ostrinia furnacalis(Guenée)等害虫具有高毒力的蛋白,具有良好的应用前景。为了明确亚洲玉米螟对Cry9Ee和Cry1Ab蛋白是否存在交互抗性,本文首先分别表达、活化、纯化了Cry9Ee和Cry1Ab蛋白,进而对敏感和Cry1Ab抗性亚洲玉米螟进行生物活性测定;随后对2种蛋白在敏感亚洲玉米螟中肠刷状缘膜囊泡(brush border membrane vesicles,简称BBMVs)上的结合进行分析比较。生物活性测定结果表明,Cry9Ee和Cry1Ab蛋白的活性片段对敏感亚洲玉米螟的LC₅₀分别为3.04和0.89 μg/g;而Cry9Ee活性片段在5.00 μg/g的浓度下,对Cry1Ab蛋白抗性亚洲玉米螟致死率高达96%,与对敏感种群活性相当。竞争结合试验表明,Cry9Ee和Cry1Ab两种蛋白间不存在竞争结合,推测它们在亚洲玉米螟中肠上存在不同的受体。综合生物活性测定及体外结合试验两方面结果得出结论：亚洲玉米螟对Cry9Ee与Cry1Ab蛋白不存在交互抗性。cry9Ee基因可以作为理想的候选基因,用于我国新一代转基因抗虫玉米的研制,为延缓和克服亚洲玉米螟对转基因抗虫玉米抗性的产生提供重要保证。

关键词: Cry9Ee蛋白, 亚洲玉米螟, 刷状缘膜囊泡, 交互抗性

Abstract: Cry9Ee, a recently discovered toxin with high insecticidal activity against Ostrinia furnacalis (Guenée), is prospective for commercialization. Cry9Ee and Cry1Ab proteins were expressed, activated and purified, and tested against susceptive and Cry1Ab-resistant O. furnacalis to verify the cross-resistance of O. furnacalis to these two proteins. The results indicated that the LC₅₀of activated Cry9Ee and Cry1Ab to susceptible O. furnacalis were 3.04 and 0.89 μg/g, respectively. At 5.00 μg/g concentration, activated Cry9Ee resulted in a 96% mortality rate against the Cry1Ab-resistant strain, which was comparable with its activity against the susceptible colony. The competition binding assay of the two proteins on BBMVs of O. furnacalis indicated no competition binding between Cry9Ee and Cry1Ab, showing that the two proteins use different receptors on the midgut of O. furnacalis. Therefore, there was no cross-resistance of O. furnacalis to Cry9Ee and Cry1Ab. Cry9Ee gene may be an ideal candidate for next generation transgenic maize that can overcome or delay resistance of insect pests to transgenic maize.

Key words: Cry9Ee protein, Ostrinia furnacalis, BBMVs, cross-resistance

中图分类号:

S476.11

王泽宇, 周子珊, 何康来, 束长龙, 宋福平, 张杰. 亚洲玉米螟对Cry9Ee与Cry1Ab蛋白的交互抗性分析[J]. 中国生物防治学报, 2015, 31(6): 882-888.

WANG Zeyu, ZHOU Zishan, HE Kanglai, SHU Changlong, SONG Fuping, ZHANG Jie. Cross-resistance of Ostrinia furnacalis to Cry9Ee and Cry1Ab Proteins[J]. journal1, 2015, 31(6): 882-888.

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