title: Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).
abstract: BACKGROUND
  Accurate evaluation of gene expression requires normalization relative to the expression of reliable reference genes. Expression levels of "classical" reference genes can differ, however, across experimental conditions. Although quantitative real-time PCR (qRT-PCR) has been used extensively to decipher gene function in the sweetpotato whitefly Bemisia tabaci, a world-wide pest in many agricultural systems, the stability of its reference genes has rarely been validated.
  
  
  RESULTS
  In this study, 15 candidate reference genes from B. tabaci were evaluated using two Excel-based algorithms geNorm and Normfinder under a diverse set of biotic and abiotic conditions. At least two reference genes were selected to normalize gene expressions in B. tabaci under experimental conditions. Specifically, for biotic conditions including host plant, acquisition of a plant virus, developmental stage, tissue (body region of the adult), and whitefly biotype, ribosomal protein L29 was the most stable reference gene. In contrast, the expression of elongation factor 1 alpha, peptidylprolyl isomerase A, NADH dehydrogenase, succinate dehydrogenase complex subunit A and heat shock protein 40 were consistently stable across various abiotic conditions including photoperiod, temperature, and insecticide susceptibility.
  
  
  CONCLUSIONS
  Our finding is the first step toward establishing a standardized quantitative real-time PCR procedure following the MIQE (Minimum Information for publication of Quantitative real time PCR Experiments) guideline in an agriculturally important insect pest, and provides a solid foundation for future RNA interference based functional study in B. tabaci.
authors: Institute of Pesticide, Hunan Agricultural University, Changsha, P. R. China.
authors: Rumei
authors: R
authors: Li
authors: Wen
authors: W
authors: Xie
authors: Shaoli
authors: S
authors: Wang
authors: Qingjun
authors: Q
authors: Wu
authors: Nina
authors: N
authors: Yang
authors: Xin
authors: X
authors: Yang
authors: Huipeng
authors: H
authors: Pan
authors: Xiaomao
authors: X
authors: Zhou
authors: Lianyang
authors: L
authors: Bai
authors: Baoyun
authors: B
authors: Xu
authors: Xuguo
authors: X
authors: Zhou
authors: Youjun
authors: Y
authors: Zhang
mesh_terms: D000818:Animals; D001937:Brassica / Q000469:parasitology / Q000821:virology; D018553:Cucumis sativus / Q000469:parasitology / Q000821:virology; D020869:Gene Expression Profiling / Q000379:methods*; D017344:Genes, Insect; D006430:Hemiptera / Q000235:genetics*; D018551:Solanum lycopersicum / Q000469:parasitology / Q000821:virology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*