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Evaluation of the reference genes for expression analysis using quantitative real‐time polymerase chain reaction in the green peach aphid, Myzus persicae
Author(s) -
Kang ZhiWei,
Liu FangHua,
Tian HongGang,
Zhang Meng,
Guo ShanShan,
Liu TongXian
Publication year - 2017
Publication title -
insect science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 45
eISSN - 1744-7917
pISSN - 1672-9609
DOI - 10.1111/1744-7917.12310
Subject(s) - myzus persicae , reference genes , biology , aphid , aphididae , gene , real time polymerase chain reaction , gene expression , genetics , ribosomal protein , pest analysis , botany , rna , homoptera , ribosome
The green peach aphid, Myzus persicae Sulzer (Hemiptera, Aphididae), is an important cosmopolitan pest. Real time qRT‐PCR has been used for target gene expression analysis on M. persicae . Using real time qRT‐PCR, the expression levels are normalized on the basis of the reliable reference genes. However, to date, the stability of available reference genes has been insufficient. In this study, we evaluated nine candidate reference genes from M. persicae under diverse experimental conditions. The tested candidate genes were comprehensively ranked based on five alternative methods (RefFinder, geNorm, Normfinder, BestKeeper and the comparative ΔC t method). 18s , Actin and ribosomal protein L27 ( L27 ) were recommended as the most stable reference genes for M. persicae , whereas ribosomal protein L27 ( L27 ) was found to be the least stable reference genes for abiotic studies (photoperiod, temperature and insecticide susceptibility). Our finding not only sheds light on establishing an accurate and reliable normalization of real time qRT‐PCR data in M. persicae but also lays a solid foundation for further studies of M. persicae involving RNA interference and functional gene research.