Selection and Validation of Reliable Reference Genes for qRT-PCR Normalization of Bursaphelenchus xylophilus from Different Temperature Conditions and Developmental Stages

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