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

        Zhen-Hong Lv,Hui-Peng Pan,Wei Zhang,Tian-Bo Ding,Dong Chu 한국응용곤충학회 2018 Journal of Asia-Pacific Entomology Vol.21 No.1

        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 referencegenes (HSP40, HSP20, HSP70, HSP90, v-ATPase, RPL29, EF-1, SDHA, Actin, PPIA, GAPDH, MyosinL,NADH, and γ-tubulin) were analyzed using five different programs including ΔCt method, BestKeeper, geNorm,NormFinder, and ReFinder to validate their use as reference genes in two invasive whiteflies, Bemisia tabaci Band Q, after acquiring the vectored virus, Tomato yellow leaf curl virus (TYLCV), or ingesting the non-vectoredvirus, Tomato spotted wilt virus (TSWV), respectively. The results showed that HSP40, v-ATPase, and EF-1 werethe most stable genes in B. tabaci B (B. tabaci B feeding on the healthy, TYLCV- and TSWV-infected tomatoplant), PPIA, 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 referencegenes 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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        Development and Application of Polymorphic SSR Markers in Luffa cylindrica

        Junjie Cui,Zhen Lv,Tianwen Yang,Jing Wang,Yu Hong 한국원예학회 2022 원예과학기술지 Vol.40 No.5

        Two published genomes of Luffa cylindrica, ‘SG2019’ and ‘SO3’, were used to develop polymorphic SSR markers for L. cylindrica by searching and comparing the SSR motifs at the same site in the two genomes. Based on the SSR search conditions and primer design criteria, 2130 polymorphic SSR markers for L. cylindrica were developed. The main motif type was dinucleotide, accounting for 80.28% of the total; the main motif units were AT/AT and AAT/ATT, accounting for 87.80% of the total. Furthermore, 40 polymorphic SSR markers developed in this study were randomly selected and amplified in 24 Luffa samples. According to the results, the 40 polymorphic SSR markers showed an amplification rate of 100% and a polymorphism rate of 80%. Cluster analysis classified the 24 Luffa samples into two main groups, L. cylindrica and L. acutangula. Overall, the polymorphic SSR markers developed in this study display a high polymorphic rate and reliable utilization value.

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