Dataset Information

Identification and validation of reference genes for gene expression analysis in Aphidius gifuensis (Hymenoptera: Aphidiidae).

ABSTRACT: Reference genes have been utilized in estimating gene expression levels using quantitative reverse transcriptase-quantitative polymerase chain reaction (qRT-PCR) analysis. Aphidius gifuensis Ashmaed is one of the most widely used biological control agents for aphids. The biological properties of this species have been studied in detail, and current investigations are focused on elucidating the regulatory mechanisms in its host However, the appropriate reference genes for target gene expression studies have not been identified. In this study, the expression profiles of 12 candidate reference genes were evaluated under different experimental conditions(development stage, sex, tissue type, diet) by using dedicated algorithms, including geNorm, Normfinder, BestKeeper, and ?Ct. In addition, RefFinder was used to rank the overall stability of the candidate genes. Finally, we recommend three optimal reference genes for the normalization of qRT-PCR data in the presence of specific variables, which include ACTB, RPL13, and PPI for different developmental stages; RPS18, ACTB, and RPL13 for sexes; RPL13, PRII3, and RPS18 in different tissue types; and RPL13, RPL27, and ACTB in diverse diets. The present study has identified optimal reference genes that could be used in estimating the expression levels of specific genes under these conditions following the Minimum Information for publication of Quantitative real-time PCR Experiments (MIQE) guidelines, which would facilitate in advancements in functional genomics research on A. gifuensis.

SUBMITTER: Gao XK

PROVIDER: S-EPMC5708624 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Identification and validation of reference genes for gene expression analysis in Aphidius gifuensis (Hymenoptera: Aphidiidae).

Gao Xue-Ke XK Zhang Shuai S Luo Jun-Yu JY Wang Chun-Yi CY Lü Li-Min LM Zhang Li-Juan LJ Zhu Xiang-Zhen XZ Wang Li L Cui Jin-Jie JJ

PloS one 20171130 11

Reference genes have been utilized in estimating gene expression levels using quantitative reverse transcriptase-quantitative polymerase chain reaction (qRT-PCR) analysis. Aphidius gifuensis Ashmaed is one of the most widely used biological control agents for aphids. The biological properties of this species have been studied in detail, and current investigations are focused on elucidating the regulatory mechanisms in its host However, the appropriate reference genes for target gene expression s ...[more]

PMID: 29190301

Similar Datasets

Project description:The integrated pest management (IPM) strategy was developed and used in combination with pesticides and beneficial biological control agents. To further develop IPM efficiency, it is important to evaluate the side effects of pesticides on biological control agents. Aphidius gifuensis is one of the most important aphid natural enemies and has been successfully used to control Myzys persicae and other aphid species. Imidacloprid (IMD) is a popular pesticide used worldwide and is highly toxic to non-target arthropods. Here, we investigated the short-term sublethal toxicity of IMD in Aphidius gifuensis and its impact on the biological performance and gene expression of this parasitoid. We found that sublethal IMD doses had a significant negative effect on the life history traits of female A. gifuensis, including shortening the lifespan and lowering parasitic capacity. Moreover, exposure to sublethal IMD also adversely affected the response of A. gifuensis to aphid-infested plant volatiles. Based on the transcriptome analysis, we found that the exposure to sublethal IMD doses significantly affected expression of genes involved in the central nervous system, energy metabolism, olfactory, and detoxification system of A. gifuensis. RT-qPCR also revealed that short term expose to sublethal IMD doses significantly induced the gene expression of genes related to the central nervous system (nAChRa7, nAChRa9, TbH, OAR1, NFR, TYR, and DAR1), olfactory system (OR28 and IR8a1), and detoxification system (CYP49p3, CYP6a2, and POD), while it suppressed the expression of genes involved in the central nervous system (nAChRa4 and nAChRb1), olfactory system (Orco1, IR8a2, and GR1), and detoxification system (GST2). Furthermore, exposure to sublethal doses of IMD also significantly increased the activities of CarEs and POD, whereas we observed no influence on the activities of CAT, GST, and SOD. Our results indicate that sublethal IMD doses might adversely affect the biological performance of A. gifuensis by altering gene expression related to the function of olfactory, nervous, energy metabolism, and detoxification systems. Thus, how the use of pesticides directly affect insect population should be considered when used in conjunction with natural pest parasitoids in IPM strategies.

Dataset Information

Identification and validation of reference genes for gene expression analysis in Aphidius gifuensis (Hymenoptera: Aphidiidae).

Publications

Identification and validation of reference genes for gene expression analysis in Aphidius gifuensis (Hymenoptera: Aphidiidae).

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets