Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5589395 | Gene | 2017 | 8 Pages |
Abstract
Lysiphlebia japonica (Ashmead) is a predominant parasitoid of cotton-melon aphids in the fields of northern China with a proven ability to effectively control cotton aphid populations in early summer. For accurate normalization of gene expression in L. japonica using quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), reference genes with stable gene expression patterns are essential. However, no appropriate reference genes is L. japonica have been investigated to date. In the present study, 12 selected housekeeping genes from L. japonica were cloned. We evaluated the stability of these genes under various experimental treatments by RT-qPCR using four independent (geNorm, NormFinder, BestKeeper and Delta Ct) and one comparative (RefFinder) algorithm. We identified genes showing the most stable levels of expression: DIMT, 18S rRNA, and RPL13 during different stages; AK, RPL13, and TBP among sexes; EF1A, PPI, and RPL27 in different tissues, and EF1A, RPL13, and PPI in adults fed on different diets. Moreover, the expression profile of a target gene (odorant receptor 1, OR1) studied during the developmental stages confirms the reliability of the chosen selected reference genes. This study provides for the first time a comprehensive list of suitable reference genes for gene expression studies in L. japonica and will benefit subsequent genomics and functional genomics research on this natural enemy.
Keywords
RPL18RPL27β-tubulinRT-qPCRRibosomal protein L27qRT-PCROBPSEF1aORFsTBPRNA polymerase II18S ribosomal RNAβ-actinArginine kinasestandard deviationelongation factor 1 alphaActbOpen reading framesNormalizationtubQuantitative reverse transcriptase-polymerase chain reactionTATA binding proteinodorant-binding proteinsReference geneolfactory receptor
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Biochemistry, Genetics and Molecular Biology
Genetics
Authors
Xue-Ke Gao, Shuai Zhang, Jun-Yu Luo, Chun-Yi Wang, Li-Min Lü, Li-Juan Zhang, Xiang-Zhen Zhu, Li Wang, Hui Lu, Jin-Jie Cui,