To correct this problem, reference genes are widely used to normalize qRT-PCR data, which can reduce error among samples and control for internal differences. Unfortunately, variation in RNA isolation, cDNA quantification, transcription, and amplification can cause qRT-PCR to be error-prone. Quantitative real-time PCR (qRT-PCR) was initially described in 1992 and has become a widely used approach to analyze gene expression due to it’s accuracy, high reproducibility and sensitivity.
chilonis, so we want to explore low temperatures to extend its shelf life so that it can be better utilized in the future. Besides, there are few studies about the storage methods of C. chilonis have been studied in previous researches, such as fecundity, parasitism rate, and sex ratio. chilonis is also an important biological control agent of some other stem borers and was previously imported into several African countries as a biological control agent. Cotesia chilonis is the dominant parasitoid of the rice stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae) larvae, which is a serious pest in China, particularly in the Yangtze River area and southern regions of China. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the manuscript.įunding: This study was supported by the National Key R&D Program of China (2017YFD0200400), the National Basic Research and Development Program of China (2013CB127604) and the Postgraduate Practical Innovation Project of Yangzhou University (XSJCX18_068).Ĭompeting interests: The authors have declared that no competing interests exist.Ĭotesia chilonis (Matsumura) (Hymenoptera: Braconidae) is native to parts of southeastern and eastern Asia. Received: Accepted: NovemPublished: December 26, 2019Ĭopyright: © 2019 Li et al. PLoS ONE 14(12):Įditor: Alberto Amato, IRIG-CEA Grenoble, FRANCE
The reference genes identified here will facilitate further investigations of the biological characteristics of this important parasitoid.Ĭitation: Li Q-Y, Li Z-L, Lu M-X, Cao S-S, Du Y-Z (2019) Selection of valid reference genes for quantitative real-time PCR in Cotesia chilonis (Hymenoptera: Braconidae) exposed to different temperatures. This study strengthens understanding of the selection of reference genes before qRT-PCR analysis in C. In samples stored at cold temperature (4☌), 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. In samples stored at normal developmental temperature (27☌), 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). The optimal numbers and stabilities of reference genes varied between the two temperature treatments (27☌ and 4☌). Data were analyzed using the ΔCt method, BestKeeper, NormFinder, and geNorm. Specifically, the performance and stabilities of these genes were compared in adult wasps maintained in a growth condition at 27☌ (normal storage conditions) and in adults obtained from pupae refrigerated at 4☌ for five days (cold storage conditions). 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.
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.