The effect of silencing arginine kinase by RNAi on the larval development of Helicoverpa armigera
The effect of silencing arginine kinase by RNAi on the larval development of Helicoverpa armigera
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RNAi沉默精氨酸激酶对棉铃虫幼虫发育的影响
DOI：
10.1017/s0007485315000450
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发表时间：
2015-10-01
影响因子：
1.9
通讯作者：
Cheng, H. -M.
中科院分区：
文献类型：
--
作者：
Qi, X. -L.;Su, X. -F.;Cheng, H. -M.
精氨酸激酶(AK)是无脊椎动物能量代谢的重要调节因子。通过cDNA扩增片段长度多态性分析，发现一个精氨酸激酶基因Haak在棉铃虫Cry1Ac敏感幼虫(96S)和抗性(Bt-R)棉铃虫幼虫中差异表达。从棉铃虫中分离到了HAAK基因的全长开放阅读框序列，其长度为1068bp。定量逆转录聚合酶链式反应分析表明，HAAK基因在多种组织和幼虫发育阶段都有特异表达。HAAK在5龄幼虫中肠中的表达水平最高。此外，HAAK基因在Bt-R幼虫中的表达明显下调。我们进一步构建了直接针对Haak的dsRNA载体，并利用RNAi技术控制了幼虫。投喂生物测定表明，微量dsRNA可显著提高幼虫死亡率，延缓幼虫化蛹。HAAK的沉默显著延缓了幼虫的发育，表明HAAK是基于RNA干扰的害虫管理的潜在靶标。
Arginine kinase (AK) is an important regulation factor of energy metabolism in invertebrate. An arginine kinase gene, named HaAK, was identified to be differentially expressed between Cry1Ac-susceptible (96S) and Cry1Ac-resistant (Bt-R) Helicoverpa armigera larvae using cDNA-amplification fragment length polymorphism analysis. The full-length open reading frame sequence of HaAK gene with 1068 bp was isolated from H. armigera. Quantitative reverse transcription polymerase chain reaction assay revealed that HaAK gene is specifically expressed in multiple tissues and at larval developmental stages. The peak expression level of HaAK was detected in the midgut of the fifth-instar larvae. Moreover, the expression of HaAK was obviously down-regulated in Bt-R larvae. We further constructed a dsRNA vector directly targeting HaAK and employed RNAi technology to control the larvae. The feeding bioassays showed that minute quantities of dsRNA could greatly increase the larval mortality and delay the larval pupation. Silencing of HaAK significantly retarded the larval development, indicating that HaAK is a potential target for RNA interference-based pest management.