Developmental control of a lepidopteran pest Spodoptera exigua by ingestion of bacteria expressing dsRNA of a non-midgut gene.
Developmental control of a lepidopteran pest Spodoptera exigua by ingestion of bacteria expressing dsRNA of a non-midgut gene.
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DOI：
10.1371/journal.pone.0006225
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发表时间：
2009-07-13
期刊：
影响因子：
3.7
通讯作者：
Zhang W
中科院分区：
文献类型：
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作者：
Tian H;Peng H;Yao Q;Chen H;Xie Q;Tang B;Zhang W
通过双链RNA（dsRNA）诱导的RNA干扰（RNAi）已被广泛应用于昆虫基因功能的研究。最近，有报道称，通过饲喂dsRNA可以成功地诱导几种昆虫的基因敲低。然而，它仍然是未知的表型沉默的基因不表达的中肠摄入昆虫dsRNA后发生。以在鳞翅目害虫甜菜夜蛾（Spodoptera exigua）角质层和气管中表达的几丁质合成酶基因A（SeCHSA）为靶基因，研究了甜菜夜蛾（Spodoptera exigua）的生长发育过程。饲喂表达SeCHSA dsRNA的大肠杆菌的甜菜幼虫受到干扰，导致致死。在4龄、5龄、蛹前期和蛹期，取食含dsRNA饲料的昆虫的平均存活率分别为88.64%、74.24%、68.43%和62.63%。5龄幼虫、预蛹期和蛹期的存活率均显著低于各对照组，4龄幼虫的致死率与对照组和ddH_2O对照组差异显著。外源细菌表达的dsRNA对目的基因的转录、组织结构和昆虫存活率的影响具有剂量依赖性。我们的研究结果表明，SeCHSA dsRNA可能是有用的昆虫害虫控制的手段。
RNA interference (RNAi) induced through double stranded RNA (dsRNA) has been used widely to study gene function in insects. Recently, it has been reported that gene knockdown in several insects can be induced successfully through feeding with dsRNA. However, it is still unknown whether phenotypic silencing of genes not expressed in the midgut occurs after ingestion of insect dsRNA. Using chitin synthase gene A (SeCHSA) as the target gene, which is expressed in the cuticle and tracheae of the lepidopteran pest Spodoptera exigua, we showed that the growth and development of S. exigua larvae fed Escherichia coli expressing dsRNA of SeCHSA was disturbed, resulting in lethality. In the 4th and 5th larval instars, prepupae, and pupae, the mean survival rates of insects fed the dsRNA-containing diet were 88.64%, 74.24%, 68.43% and 62.63% respectively. The survival rates in the 5th instar larvae, prepupae and pupae stages were significantly lower than those of all controls, and significant lethality differences were also found between dsSeCHSA treatment and dsControl or ddH2O control in the 4th instar larvae. The effects of ingesting bacterially expressed dsRNA on transcription of the target gene, tissue structure, and survival rates of insects were dose-dependent. Our results suggest that SeCHSA dsRNA may be useful as a means of insect pest control.
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