Selection of stable reference genes for accurate reverse-transcription quantitative PCR in cotton-herbivore studies using virus-induced gene silencing.

ABSTRACT: Virus-induced gene silencing (VIGS) has been a crucial tool for elucidating gene function in Upland cotton (Gossypium hirsutum) due to its complex allotetraploid genome. Reverse-transcription quantitative PCR is routine for measuring gene expression in VIGS studies, yet reference gene stability has not been adequately evaluated when using VIGS especially under biotic stress in cotton. Here, we employed several statistical methods (∆Ct, geNorm, BestKeeper, NormFinder, and weighted rank aggregation) to evaluate the stability of six candidate reference genes (GhACT7, GhPP2A1, GhUBQ7, GhUBQ14, GhTMN5, and GhTBL6) in wild-type and VIGS-infiltrated plants under cotton aphid herbivory stress over time in a fully factorial experiment. Ranked stability analyses overwhelmingly indicated that the fre

SUBMITTER: Clark MK 

PROVIDER: S-EPMC12238633 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature