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Reference gene selection for RT-qPCR analysis in two invasive whiteflies after the acquisition of vectored or non-vectored viruses

文献类型： 外文期刊

作者： Lv, Zhen-Hong 1 ; Pan, Hui-Peng 2 ; Zhang, Wei 3 ; Ding, Tian-Bo 1 ; Chu, Dong 1 ;

作者机构： 1.Qingdao Agr Univ, Coll Plant Protect, Key Lab Integrated Crop Pest Management Shandong, Qingdao 266109, Peoples R China

2.South China Agr Univ, Dept Entomol, Key Lab Biopesticide Innovat & Applicat Guangdong, Guangzhou 510642, Guangdong, Peoples R China

3.Shandong Acad Agr Sci, Jinan 250100, Shandong, Peoples R China

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

期刊名称：JOURNAL OF ASIA-PACIFIC ENTOMOLOGY （ 影响因子：1.303； 五年影响因子：1.427 ）

ISSN： 1226-8615

年卷期： 2018 年 21 卷 1 期

页码：

收录情况： SCI

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.

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