Selection of reference genes for RT-qPCR analysis of Phenacoccus solenopsis (Hemiptera: Pseudococcidae) sex-dimorphic development
| Autor: | Zhijun Zhang, Xiao-wei Li, Chuan-ren Li, Huang Jun, Wencai Lin, Jin-ming Zhang, Wei-di Li, Yao-bin Lu, Lu-ying Zheng |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0106 biological sciences
media_common.quotation_subject Agriculture (General) reference gene Plant Science Insect 01 natural sciences Biochemistry sexual dimorphic development S1-972 gene stability Food Animals Reference genes Gene expression Gene Selection (genetic algorithm) media_common Genetics Ecology biology RT-qPCR 04 agricultural and veterinary sciences biology.organism_classification Hemiptera Sexual dimorphism Phenacoccus solenopsis Juvenile hormone 040103 agronomy & agriculture 0401 agriculture forestry and fisheries Animal Science and Zoology Agronomy and Crop Science 010606 plant biology & botany Food Science |
| Zdroj: | Journal of Integrative Agriculture, Vol 18, Iss 4, Pp 854-864 (2019) |
| ISSN: | 2095-3119 |
| Popis: | Mealybugs, such as Phenacoccus solenopsis, are highly sexually dimorphic. Winged adult males present such remarkable morphological differences from females that, to the untrained eye, conspecific adults of both sexes of P. solenopsis may be considered as two different insect species. A method to investigate sex-dimorphic mechanisms is by evaluating gene expression using RT-qPCR. However, the accuracy and consistency of this technique depend on the reference gene(s) selected. In this study, we analyzed the expression of 10 candidate reference genes in male and female P. solenopsis at different development stages, using common algorithms including the ΔCt method, NormFinder, geNorm, BestKeeper, and a web-based analysis tool, RefFinder. The results showed that EF1-β, RP-L32 and RP-18S were selected as the most stable genes by both the ΔCt method and NormFinder; TUB-α was the most stable gene identified by BestKeeper; and RP-L40 and RP-L32 were the most stable genes ranked by geNorm. RefFinder, a comprehensive analysis software, ranked the ten genes and determined EF1-β and RP-L32 as the most suitable reference genes for the various developmental stages in male and female P. solenopsis. Furthermore, the two most suitable reference genes were validated by examining expression of the juvenile hormone acid O-methytransferase (JHAMT) gene. Results of the validation portion of the study showed that JHAMT expression was sex-biased towards males and exhibited a dynamic and classic expression pattern among the P. solenopsis developmental stages. The results can help further our knowledge on the molecular mechanisms underlying sexual dimorphic development in P. solenopsis. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect