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Screening of reference genes using real-time quantitative PCR for gene expression studies in Neoseiulus barkeri Hughes (Acari: Phytoseiidae)

Published online by Cambridge University Press:  29 October 2018

C. Wang ,
J. Yang ,
Q. Pan ,
S. Yu ,
R. Luo ,
H. Liu ,
H. Li ,
L. Cong  and
C. Ran
C. Wang
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
J. Yang
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
Q. Pan
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
S. Yu
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
R. Luo
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
H. Liu
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
H. Li
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
L. Cong
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
C. Ran*
Affiliation:
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, China
*
*Author for correspondence Phone: +86-23-6834-9798 Fax: 023-68349005 E-mail: ranchun@cric.cn
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Abstract

A stable reference gene is a key prerequisite for accurate assessment of gene expression. At present, the real-time reverse transcriptase quantitative polymerase chain reaction has been widely used in the analysis of gene expression in a variety of organisms. Neoseiulus barkeri Hughes (Acari: Phytoseiidae) is a major predator of mites on many important economically crops. Until now, however, there are no reports evaluating the stability of reference genes in this species. In view of this, we used GeNorm, NormFinder, BestKeeper, and RefFinder software tools to evaluate the expression stability of 11 candidate reference genes in developmental stages and under various abiotic stresses. According to our results, β-ACT and Hsp40 were the top two stable reference genes in developmental stages. The Hsp60 and Hsp90 were the most stable reference genes in various acaricides stress. For alterations in temperature, Hsp40 and α-TUB were the most suitable reference genes. About UV stress, EF1α and α-TUB were the best choice, and for the different prey stress, β-ACT and α-TUB were best suited. In normal conditions, the β-ACT and α-TUB were the two of the highest stable reference genes to respond to all kinds of stresses. The current study provided a valuable foundation for the further analysis of gene expression in N. barkeri.

Keywords

RT-qPCRNeoseiulus barkerireference genesgene expression
Type
Research Papers
Information
Bulletin of Entomological Research , Volume 109 , Issue 4 , August 2019 , pp. 443 - 452
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Copyright
Copyright © Cambridge University Press 2018 

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Footnotes

†

These authors contributed equally to this study.

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