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2×SYBR realtime RT-PCR premixture2
Reference Gene Selection for qRT-PCR Analysis in the
Sweetpotato Whitefly, Bemisia tabaci (Hemiptera:
Aleyrodidae)
Rumei Li1,2, Wen Xie2, Shaoli Wang2, Qingjun Wu2, Nina Yang2, Xin Yang2, Huipeng Pan2,3,
Xiaomao Zhou1, Lianyang Bai1, Baoyun Xu2, Xuguo Zhou3*, Youjun Zhang2*
1 Institute of Pesticide, Hunan Agricultural University, Changsha, P. R. China, 2 Department of Plant Protection, Institute of Vegetables and Flowers, Chinese Academy of
Agricultural Sciences, Beijing, P. R. China, 3 Department of Entomology, University of Kentucky, Lexington, Kentucky, United States of America
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.
Conclusion: Our finding is the first step toward establishing a standardized quan
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