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
第一作者: Lv, Zhen-Hong
作者: Lv, Zhen-Hong;Ding, Tian-Bo;Chu, Dong;Pan, Hui-Peng;Zhang, Wei
作者机构:
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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