莲草直胸跳甲羧酸酯酶基因AhCesB1的克

发布时间：2018-08-11 20:04

【摘要】：莲草直胸跳甲Agasicles hygrophila是入侵杂草喜旱莲子草的专食性天敌。为进一步明确莲草直胸跳甲寄主专一性的分子机制,通过分析其转录组数据克隆羧酸酯酶(carboxylesterases,Car Es)基因,采用RT-PCR技术从其体内获得1个羧酸酯酶B簇基因并进行生物信息学分析,同时利用实时定量PCR方法检测其在不同身体部位及不同发育阶段的表达特性,并对饥饿处理、取食靶标植物喜旱莲子草和非靶标植物儥僘后该基因的表达水平进行比较。结果显示,克隆得到1条莲草直胸跳甲羧酸酯酶基因的cDNA全长序列,长度为1 851 bp,开放阅读框长度为1 647 bp,编码548个氨基酸,以该基因推导的氨基酸序列与其它物种的CarEs进行聚类分析,发现此基因为羧酸酯酶B簇家族成员,命名为AhCesB1(GenBank登录号KT032151)。实时定量PCR检测结果显示,AhCesB1基因在莲草直胸跳甲3龄期表达量最高,成虫期和蛹期表达量次之,卵、1龄和2龄期表达量相对较低。AhCesB1基因在头和中肠的表达量较低,在其余身体部分表达量较高。取食儥僘后AhCesB1表达量显著增高,为饥饿处理的13.7倍,而取食喜旱莲子草与饥饿处理并无显著差异。表明莲草直胸跳甲AhCesB1在不同发育阶段和成虫不同身体部位具有不同的表达水平,而非寄主植物儥僘可以诱导该基因的表达,推测羧酸酯酶在莲草直胸跳甲代谢次生物质中发挥着重要作用。
[Abstract]:A gasicles hygrophila is the special natural enemy of the invasive weed. In order to further clarify the molecular mechanism of host specificity, carboxylesterase (carboxylesterase) gene was cloned by analyzing its transcriptional data, and a carboxylesterase B cluster gene was obtained by RT-PCR technique and analyzed by bioinformatics. At the same time, real-time quantitative PCR was used to detect the expression characteristics of the gene in different body parts and different developmental stages, and the expression level of the gene was compared between the starvation treatment, feeding on the target plant and the non-target plant. The results showed that the full-length cDNA sequence of a straight leucosternase gene was 1 851 BP, and the open reading frame length was 1 647 BP, encoding 548 amino acids, the total length of the cDNA was 1 851 BP, and the length of the open reading frame was 1 647 BP, which encoded 548 amino acids. The deduced amino acid sequence of the gene was clustered with the CarEs of other species. It was found that this gene was named AhCesB1 (GenBank accession number KT032151) because of the carboxylesterase B family member. The results of real-time quantitative PCR analysis showed that the expression of AhCesB1 gene was the highest in the 3rd age period, followed by the adult stage and pupa stage. The expression of AhCesB1 gene in the head and midgut was relatively low in the 1st and 2nd instar of oocyte. In the rest of the body part of the expression is higher. The expression of AhCesB1 was 13.7 times higher than that of the starvation treatment, but there was no significant difference between the feeding and the starvation treatment. The results indicated that the expression level of A hCesB1 was different in different developmental stages and different body parts of adults, and the expression of the gene could be induced by the non-host plant. It is speculated that carboxylesterase plays an important role in the metabolism of secondary biomass.
【作者单位】：山西农业大学农学院;
【基金】：国家自然科学基金(31301723) 中国博士后科学基金(2013M541204) 山西农业大学专业提升计划(TSJH1406)
【分类号】：S451;S476

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