FUNCTIONAL ANALYSIS OF TWO CHITINASE GENES DURING THE PUPATION AND ECLOSION STAGES OF THE BEET ARMYWORM Spodoptera exigua BY RNA INTERFERENCE
FUNCTIONAL ANALYSIS OF TWO CHITINASE GENES DURING THE PUPATION AND ECLOSION STAGES OF THE BEET ARMYWORM Spodoptera exigua BY RNA INTERFERENCE
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DOI：
10.1002/arch.21018
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发表时间：
2012-04-01
影响因子：
2.2
通讯作者：
Zhang, Wenqing
中科院分区：
文献类型：
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作者：
Zhang, Daowei;Chen, Jing;Zhang, Wenqing
昆虫几丁质酶是一个多基因家族，由一个相当大的和多样化的基因组编码。几丁质酶的主要功能是在蜕皮期间消化包含在组织如角质层和肠衬里中的几丁质。本文研究了几丁质酶（SeChi）和细菌型几丁质酶（SeChi-h）在甜菜夜蛾化蛹和羽化过程中的作用。首先，通过特异性双链RNA（dsRNA）注射有效沉默SeChi和SeChi-h基因导致SeChi和SeChi-h的mRNA水平显著降低。此外，在注射SeChi和SeChi-h dsRNA后，在蛹和成虫阶段观察到不同的表型缺陷。在蛹期注射SeChi dsRNA后，头部的角质层裂开，并且蛹角质层在旧幼虫角质层下可见。然而，在注射SeChi-h dsRNA后，动物死亡，没有表现出任何特殊的表型。在成虫死亡阶段，注射dsSeChi的动物不能完全脱落它们的蛹壳，并且它们的旧蛹仍然附着在它们的头部或胸部。然而，主要致死表型是在dsSeChi-h注射后昆虫不出现。此外，S.注射SeChi dsRNA后，蛹期和成虫期对甜菜夜蛾的杀虫效果分别为52.02%和40.38%。注射SeChi-h dsRNA的昆虫存活率分别为72.38%和48.52%。这些结果表明，SeChi和SeChi-h在蛹-成虫蜕皮过程中可能具有不同的生物学功能。(C)2012 Wiley Periodicals，Inc.
Insect chitinases are a multigene family that is encoded by a rather large and diverse group of genes. The main function of chitinases is to digest the chitin contained in tissues such as the cuticles and gut lining during molting. In this study, we examined the role of a chitinase (SeChi) and a bacterial type chitinase (SeChi-h) during the pupation and eclosion stages of Spodoptera exigua. First, efficient silencing of the SeChi and SeChi-h genes through specific double-stranded RNA (dsRNA) injection led to a significant reduction in the mRNA levels of SeChi and SeChi-h. Additionally, different phenotypic defects were observed at the pupal and adult stages after injection of the SeChi and SeChi-h dsRNAs. After injecting SeChi dsRNA in the pupal stage, the cuticle of the head split open and the pupal cuticle was visible under the old larval cuticle. However, after injecting the SeChi-h dsRNA, animals died without exhibiting any special phenotypes. At the adult death stage, animals injected with dsSeChi could not shed their pupal shell completely, and their old cuticles remained attached to their head or chest. However, the main lethal phenotype was that insects did not emerge after dsSeChi-h injection. Additionally, the average survival rates of S. exigua were 52.02% and 40.38% at the pupal and adult stages, respectively, after injection with SeChi dsRNA. For the insects injected with SeChi-h dsRNA, the survival rates were 72.38% and 48.52%, respectively. These results suggest that SeChi and SeChi-h may have different biologic functions during the pupaladult molting. (C) 2012 Wiley Periodicals, Inc.