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斜纹夜蛾(鳞翅目:夜蛾科)中两个乙酰胆碱酯酶基因的克隆、表达及功能分析

Cloning, expression, and functional analysis of two acetylcholinesterase genes in Spodoptera litura (Lepidoptera: Noctuidae).

作者信息

Salim Abdalla M A, Shakeel Muhammad, Ji Jinyun, Kang Tinghao, Zhang Yashu, Ali Ehsan, Xiao Zhao, Lu Yanhui, Wan Hu, Li Jianhong

机构信息

Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, PR China.

Hubei Insect Resources Utilization and Sustainable Pest Management Key Laboratory, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, PR China; State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, PR China.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2017 Apr;206:16-25. doi: 10.1016/j.cbpb.2017.01.007. Epub 2017 Jan 19.

Abstract

Two acetylcholinesterase genes (SlAce1 and SlAce2) were cloned from Spodoptera litura, which is an important pest that causes widespread economic damage to vegetables and ornamental plants. We analyzed their expression patterns and compared their biological functions by using RNA interference. Our results showed that SlAce1 and SlAce2 cDNA contains 2085bp and 1917bp nucleotides and encoding proteins of 694 and 638 amino acid residues, respectively. Phylogenic analysis indicated that the lineage of SlAce genes and SlAce1 was completely different from SlAce2. Although both genes were expressed in all developmental stages and majorly in the brain. The expression levels of the both genes were suppressed by inserting their related dsRNA in the 6th instar larvae, which led to 47.3% (SlAce1) and 37.9% (SlAce2) mortality. Interestingly, the suppression of the SlAce2 transcripts also led to significant reductions in the fecundity, hatching, and offspring in the parental generation of S. litura. It is concluded that SlAce2 is responsible for the hydrolysis of acetylcholine and also plays role in female breeding, embryo progress, and the development of progeny. Considerable larval mortality was observed after both AChE genes (i.e. Ace1 and Ace2) were silenced in S. litura confirms its insecticidal effectiveness, which provided a molecular basis in biological pest control approach.

摘要

从斜纹夜蛾中克隆了两个乙酰胆碱酯酶基因(SlAce1和SlAce2),斜纹夜蛾是一种对蔬菜和观赏植物造成广泛经济损害的重要害虫。我们分析了它们的表达模式,并通过RNA干扰比较了它们的生物学功能。我们的结果表明,SlAce1和SlAce2 cDNA分别包含2085bp和1917bp核苷酸,编码的蛋白质分别为694和638个氨基酸残基。系统发育分析表明,SlAce基因家族中SlAce1与SlAce2完全不同。尽管这两个基因在所有发育阶段均有表达,且主要在脑中表达。在6龄幼虫中插入它们的相关双链RNA可抑制这两个基因的表达水平,这导致了47.3%(SlAce1)和37.9%(SlAce2)的死亡率。有趣的是,抑制SlAce2转录本也导致斜纹夜蛾亲代的繁殖力、孵化率和后代数量显著降低。结论是,SlAce2负责乙酰胆碱的水解,并且在雌性繁殖、胚胎发育和子代发育中也发挥作用。在斜纹夜蛾中沉默两个乙酰胆碱酯酶基因(即Ace1和Ace2)后观察到相当高的幼虫死亡率,证实了其杀虫效果,这为生物防治害虫方法提供了分子基础。

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