Quantitative reverse transcription polymerase chain reaction (qRT-PCR) is a powerful technique for studying gene expression. The key to quantitative accuracy depends on the stability of the reference genes used for data normalization under different experimental conditions. Pine wood nematode (PWN, Bursaphelenchus xylophilus) is the causal agent of the devastating pine wilt disease (PWD). Extensive and prompt research is needed to understand the molecular mechanism of PWD, but identification of the reference PWN genes for standardized qRT-PCR has not been reported yet. We have analyzed eight candidate reference genes of PWN across different temperature conditions and developmental stages. Delta Ct method, GeNorm, NormFinder, BestKeeper, and RefFinder algorithms were used to evaluate the stability of expression of these genes. Finally, we use heat shock protein 90 (HSP90) in different temperatures and arginine kinase gene (AK) in different developmental stages to confirm the stability of these genes. UBCE and EF1γ were most stable across different temperature treatments, whereas EF1γ and Actin were most stable across different developmental stages. In general, these results indicate that EF1γ is the most stable gene for qRT-PCR under different conditions. The systematic analysis of qRT-PCR reference gene selection will be helpful for future functional analysis and exploration of B. xylophilus genetic resources.

邻苯二甲酸丁基苄酯(BBP)是邻苯二甲酸酯的一种,作为常用的增塑剂和溶剂,广泛应用于医疗、化工、食品、装修材料等产品,并在环境介质及人体中广泛检出. BBP被认为具有类雌激素作用,具有一定的生殖发育毒性.本文主要对BBP的代谢暴露水平、BBP及其主要代谢产物在雄性生殖发育毒性方面的研究进展进行了综述.