Abstract
In quantitative real-time PCR (qRT-PCR), data are normalized using reference genes, which helps to control for internal differences and reduce error among samples. In this study, the expression profiles of eight candidate housekeeping genes, 18S ribosomal (18S rRNA), elongation factor (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein L10 (RPL10), ribosomal protein L17 (RPL17), histone 3 (H3), arginine kinase (AK), amd β-Actin (ACTB), were evaluated in the parasitic wasp Cotesia chilonis in response to different temperatures. Specifically, the performance and stabilities of these genes were compared in adult wasps maintained in a growth condition at 27°C (normal storage conditions) and in adults obtained from pupae refrigerated at 4°C for five days (cold storage conditions). Data were analyzed using the ΔCt method, BestKeeper, NormFinder, and geNorm. The optimal numbers and stabilities of reference genes varied between the two temperature treatments (27°C and 4°C). In samples stored at normal developmental temperature (27°C), the requirement for normalization in response to low temperature exposures was three genes (18S, H3, AK), whereas normalization in response to high temperature exposures required only two reference genes (GAPDH, ACTB). In samples stored at cold temperature (4°C), for low temperature exposures two reference genes (RPL17, RPL10) were required for standardization, while following high temperature exposures three reference genes (18S, H3, ACTB) were needed. This study strengthens understanding of the selection of reference genes before qRT-PCR analysis in C. chilonis. The reference genes identified here will facilitate further investigations of the biological characteristics of this important parasitoid.
Highlights
Cotesia chilonis (Matsumura) (Hymenoptera: Braconidae) is native to parts of southeastern and eastern Asia [1, 2]
We evaluated eight candidate reference genes (18S rRNA, ACTB, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), EF1, histone 3 (H3), ribosomal protein L17 (RPL17), ribosomal protein L10 (RPL10), and arginine kinase (AK)) for expression stability in C. chilonis samples stored either at 27 oC or 4 oC and subjected to different temperatures
The reference genes identified in this study provide useful information on the ecological adaptation of this parasitoid wasp and will promote future research of gene expression
Summary
Cotesia chilonis (Matsumura) (Hymenoptera: Braconidae) is native to parts of southeastern and eastern Asia [1, 2]. Cotesia chilonis is the dominant parasitoid of the rice stem borer, Chilo. C. chilonis is an important biological control agent of some other stem borers and was previously imported into several African countries as a biological control agent [3, 4, 5]. Related biological characteristics of C. chilonis have been studied in previous researches, such as fecundity, parasitism rate, and sex ratio [6, 7]. There are few studies about the storage methods of C. chilonis, so we want to explore low temperatures to extend its shelf life so that it can be better utilized in the future
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
