蜡蚧轮枝菌转几丁质酶基因高毒力重组菌株对茶蚜的毒力
首发时间:2017-05-11
摘要:本文利用基因重组技术超量表达几丁质酶,将蜡蚧轮枝菌Vl6063、V3450、Vp28菌株的几丁质酶基因ChitVl6063、ChitV3450、ChitVp28的开放阅读框(ORF)序列克隆到含有强启动子、终止子的真菌表达载体pBARGPE-1,构建几丁质酶超量表达载体。转化野生型菌株芽生孢子,得到重组菌株。3株重组菌株的几丁质酶活性相对于野生型菌株分别提高了66.2%,63.5%,63.0%。在茶蚜无翅成虫生物测定中,转基因菌株的毒力明显高于野生型菌株,LC50降低至原来的3.43%、1.72%、1.23%;LT50缩短了29.51%、30.46%、33.90%。表明几丁质酶超量表达的转基因菌株,其毒力得到了显著提高。为构建工程菌株及提高杀虫菌剂毒力提供了依据。
关键词: 虫生真菌 蜡蚧轮枝菌 几丁质酶 转基因 超量表达 毒力测定。
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Transformants of Lecanicillium lecanii overexpressing chitinase increase virulence against Toxoptera aurantii
Abstract:In this paper, gene recombination technology was used to express Chitinase of three strains of Lecanicillium lecanii Vl6063、V3450、Vp28. The ORF of ChitVl6063, ChitV3450 and ChitVp28 were inserted into the fungal expression vector pBARGPE-1 which contain strong promoter and terminator respectively to construct a chitinase overpressing plasmid, then transformed the wild-type strain with blastospore transformation method. Insect bioassay revealed that overproduction of ChitVl6063, ChitV3450 and ChitVp28 enhanced the virulence of the three transgenic strains against Ttoxoptera aurantii, as indicated by significantly lower 3.43%, 1.72% and 1.23% lethal concentrations, respectively, and 29.51%, 30.46% and 33.90% lethal times respectively, of the transformants compared to the values for the wild-type strain.
Keywords: Entomopathogenic fungi Lecanicillium (Verticillium) lecanii chitinase transgene overexpression bioassay of toxicity
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蜡蚧轮枝菌转几丁质酶基因高毒力重组菌株对茶蚜的毒力
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