热胁迫下棉花粉蚧内参基因的筛选

【目的】筛选出热胁迫下棉花粉蚧Phenacoccus solenopsis的实时定量PCR最适内参基因。【方法】 本研究应用实时荧光定量PCR技术测定了棉花粉蚧2龄若虫、3龄若虫和雌成虫α-tub, β-tub, rpl32, GAPDH, SDHA和TBP共6个候选内参基因在7个不同温度处理（恒温18℃和 32℃, 以及37℃, 39℃, 41℃, 43℃和45℃热激处理1 h并在26℃恢复1 h）下mRNA表达水平的稳定性；应用geNorm, Bestkeeper, Normfinder和RefFinder软件分析6个基因表达的稳定性。【结果】 在不同高温胁迫下2龄若虫6个候选内参基因的稳定值M由小到大依次为α-tub（0.579）< GAPDH（0.654)< TBP（0.663）<β-tub（0.668）< rpl32（0.675）

Selection of reference genes in Phenacoccus solenopsis (Hemiptera: Pseudococcidae) under heat stress

【Aim】 To select the suitable reference genes in Phenacoccus solenopsis after exposure to different temperature stress by using quantitative real-time PCR (RT-qPCR). 【Methods】 In this study, we investigated the mRNA expression stability of six candidate reference genes (α-tub, GAPDH, rpl32, β-tub, SDHA and TBP) in different developmental stages (2nd instar nymph, 3rd instar nymph and female adult) of P. solenopsis under seven different temperature regimes (constant temperature 18℃ and 32℃, and temporary heat shock (1 h) at 37℃, 39℃, 41℃, 43℃ and 45℃ and then recovery at 26℃ for 1 h) by RT-qPCR method. Three software-based approaches (geNorm, Bestkeeper and Normfinder) and one web-based comprehensive tool (RefFinder) were used to analyze and rank the tested genes, and evaluate the expression stability of these genes. 【Results】 In the 2nd instar nymphs, the M values of six reference genes stability in the ascending order were α-tub (0.579)