摘要
RNA干扰(RNAi)不但可以用于研究基因的功能,还可以通过沉默靶标基因干扰特定的生命过程。因此,通过深入研究,发掘高效专一性靶基因和RNAi技术,有可能开辟针对性的害虫RNAi防控新途径。本研究通过灰飞虱Laodelphax striatellus转录组数据分析并结合RACE技术,克隆了灰飞虱两种海藻糖酶的全长基因,分别命名为LSTre-1和LSTre-2,其GenBank登录号分别为JQ027050和JQ027051。它们均具有海藻糖酶基因的典型特征,与已报道的其他昆虫的海藻糖酶基因具有很高的相似性,并表现出一定的虫种亲缘关系。其中LSTre-1为水溶性海藻糖酶基因,全长2042 bp,开放阅读框编码602个氨基酸,前端有25个氨基酸的信号肽,但无跨膜结构域;LSTre-2为膜结合型海藻糖酶基因,全长2619 bp,开放阅读框编码618个氨基酸,前端有26个氨基酸的信号肽,有2个疏水性跨膜结构域。利用喂食法研究2种海藻糖酶基因dsRNA对灰飞虱的致死效应,发现靶向水溶性酶基因的干扰效应略高于靶向膜结合型的,但两种海藻糖酶基因的dsRNA都可以显著抑制灰飞虱海藻糖酶基因的表达,降低其活力,还能显著抑制试虫的生长,大幅增加试虫死亡率。结果提示,通过适宜途径干扰海藻糖酶基因可以开发防治灰飞虱的新途径。
RNA interference (RNAi) has been demonstrated to be used to interfere special life processes by silencing target genes. Therefore,a new countermeasure of RNAi pest control could be developed through exploring effective specific target genes and efficient RNAi techniques. Based on transcriptome data analysis and RACE strategy,two full-length trehalase genes were cloned from the small brown plant hopper (SBPH) ,Laodelphax striatellus,and were designated as LSTre-1 and LSTre-2,with GenBank accession numbers JQ027050 and JQ027051,respectively. Both LSTre-1 and LSTre-2 have typical characteristics of trehalase genes,and show high sequence identity with trehalase genes in other insects. LSTre-1 is 2 024 bp in length,and its ORF encodes a protein of 602 amino acids including a signal peptide of 25 amino acids and without hydrophobic transmembrane domains. LSTre-2 is 2 619 bp in length,and its ORF encodes a protein of 618 amino acids including a signal peptide of 26 amino acids and two hydrophobic transmembrane domains. Double-stranded RNA (dsRNA) was synthesized based on these two genes and the toxicological effects of RNAi were studied. The results showed that the silencing effect of LSTre-1 dsRNA was higher than that of LSTre-2 dsRNA. However,silencing of these two genes all can not only cause lethal effect by decreasing the mRNA expression and enzyme activity but also prevent the normal growth and development of SBPH. These results imply that SBPH could be controlled through silencing its trehalase genes by feeding dsRNA with proper ways.
出处
《昆虫学报》
CAS
CSCD
北大核心
2012年第8期911-920,共10页
Acta Entomologica Sinica
基金
国家自然科学基金项目(30771428)
