中文摘要：定制内参基因选择对于不同种类昆虫和各种实验条件下的逆转录酶-定量聚合酶链反应（RT-qPCR）是必不可少的。本研究使用5种不同的程序（包括Delta Ct method，BestKeeper，geNorm，NormFinder和ReFinder）分析了14种候选内参基因（(HSP40，HSP20，HSP70，HSP90，v-ATPase，RPL29，EF-1，SDHA，Actin，PPM，GAPDH，MyosinL，NADH和gamma-tubulin），以确认它们是否可以分别作为两种侵入性粉虱即B型烟粉虱和Q型烟粉虱在获取载体病毒番茄黄化曲叶病毒（TYLCV）或摄取非载体病毒番茄斑萎病毒(TSWV)后的内参基因。研究结果表明，HSP40，v-ATPase和EF-1是B型烟粉虱中最为稳定的基因（B型烟粉虱取食健康的、感染TYLCV和TSWV的番茄植株），PPLA，SDHA和RPL29是Q型烟粉虱中最为稳定的基因（Q型烟粉虱取食健康的、感染TYLCV和TSWV的番茄植株）。此外，EF-1，RPL29和HSP20是B型和Q型烟粉虱中最稳定的内参基因。这些发现为将来基于RT-qPCR的烟粉虱与病毒间交互作用研究提供了基础。同时，该报道可能为摄入非载体病毒后昆虫内参基因选择开创了先例。
外文摘要：Custom reference gene selection is essential for reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) in different species of insects and various experiment conditions. In this study, 14 candidate reference genes (HSP40, HSP20, HSP70, HSP90, v-ATPase, RPL29, EF-1, SDHA, Actin, PPM, GAPDH, MyosinL, NADH, and gamma-tubulin) were analyzed using five different programs including Delta Ct method, BestKeeper, geNorm, NormFinder, and ReFinder to validate their use as reference genes in two invasive whiteflies, Bemisia tabaci B and Q, after acquiring the vectored virus, Tomato yellow leaf curl virus (TYLCV), or ingesting the non-vectored virus, Tomato spotted wilt virus (TSWV), respectively. The results showed that HSP40, v-ATPase, and EF-1 were the most stable genes in B. tabaci B (B. tabaci B feeding on the healthy, TYLCV- and TSWV-infected tomato plant), PPLA, SDHA, and RPL29 were the most stable genes in B. tabaci Q (B. tabaci Q feeding on the healthy, TYLCV- and TSWV-infected tomato plant). In addition, EF-1, RPL29, and HSP20 were the most stable reference genes in B. tabaci B and Q. These findings provide the basis for future RT-qPCR-based studies on whitefly-virus interactions. Meanwhile, this report may set a precedent for reference gene selection in insects after the ingestion of non-vectored viruses.
外文关键词：Reference gene; Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR); Bemisia tabaci B and Q; Tomato yellow leaf curl virus; Tomato spotted wilt virus
作者：Lv, Zhen-Hong; Pan, Hui-Peng; Zhang, Wei; 等
作者单位：青岛农业大学
期刊名称：JOURNAL OF ASIA-PACIFIC ENTOMOLOGY
期刊影响因子：0.875
出版年份：2018
出版刊次：3
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