High mortality caused by high dose of dsRNA in the green rice leafhopper Nephotettix cincticeps (Hemiptera: Cicadellidae)
High mortality caused by high dose of dsRNA in the green rice leafhopper Nephotettix cincticeps (Hemiptera: Cicadellidae)
复制标题
高剂量 dsRNA 导致绿叶蝉黑尾叶蝉（半翅目：叶蝉科）高死亡率
DOI：
10.1007/s13355-013-0211-5
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发表时间：
2013
影响因子：
1.3
通讯作者：
Tomizawa M and Noda H
中科院分区：
文献类型：
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作者：
Tomizawa M;Noda H;Tomizawa M and Noda H
RNA干扰(RNAi)是研究非模式昆虫和害虫基因功能的有用工具。RNAi的实验条件因昆虫不同而不同，对于评价dsRNA注射相关基因的效果是重要的。将dsRNA注入稻纵卷叶蝉NePhoettix cincticep(Uhler)体内，观察到较高的死亡率。因此，研究了注射dsRNA对叶蝉发育的不利影响，以评估在该物种中进行RNAi的适宜条件。用玻璃毛细管注射处理不影响叶蝉的存活，但延缓了相应龄期的蜕皮。当给予大量dsRNA时，观察到高死亡率。在外源性EGFP基因和内源性肽聚糖识别蛋白基因(NcPGRP12)两个基因中检测到dsRNA的这种不良反应。注射高剂量(60 ng/只)在所测的所有阶段：4龄、5龄和雌性成虫的死亡率都很高。相对较低的剂量(6 ng/只昆虫)并不会导致高死亡率，保持了很高的基因沉默潜力。由于RNAi在该物种中非常有效，因此通过给予较低剂量的dsRNA可以避免大量dsRNA的有害影响。
RNA interference (RNAi) is a useful tool for gene functional studies in non-model insects and pest insects. Establishment of experimental conditions for RNAi, which differ from insect to insect, is important for evaluating the effect of dsRNA injection of relevant genes. When injecting dsRNA into the green rice leafhopperNephotettix cincticeps(Uhler), high mortality was observed. Therefore, the adverse effects of injection of dsRNA on leafhopper development were examined to assess the suitable conditions for RNAi in this species. Injection manipulation by using a glass capillary did not affect leafhopper survival but delayed the molt of the corresponding instar. High mortality was observed when large amounts of dsRNA were administered. This adverse effect of dsRNA was examined in 2 genes, exogenousEGFPgene and endogenous peptidoglycan recognition protein gene (NcPGRP12). Injection of a high dose (60 ng/insect) caused high mortality in all stages tested: 4th instar, 5th instar, and female adult. A relatively low dose (6 ng/insect) did not cause high mortality, retaining a high potential for gene silencing. Since RNAi is highly effective in this species, the deleterious effect of large amounts of dsRNA could be avoided by administering a low dsRNA dose.