Cargando…

Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae)

Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is a reliable technique for quantifying gene expression across various biological processes, of which requires a set of suited reference genes to normalize the expression data. Coleomegilla maculata (Coleoptera: Coccinellidae), i...

Descripción completa

Detalles Bibliográficos
Autores principales: Yang, Chunxiao, Pan, Huipeng, Noland, Jeffrey Edward, Zhang, Deyong, Zhang, Zhanhong, Liu, Yong, Zhou, Xuguo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4674751/
https://www.ncbi.nlm.nih.gov/pubmed/26656102
http://dx.doi.org/10.1038/srep18201
_version_ 1782404943551922176
author Yang, Chunxiao
Pan, Huipeng
Noland, Jeffrey Edward
Zhang, Deyong
Zhang, Zhanhong
Liu, Yong
Zhou, Xuguo
author_facet Yang, Chunxiao
Pan, Huipeng
Noland, Jeffrey Edward
Zhang, Deyong
Zhang, Zhanhong
Liu, Yong
Zhou, Xuguo
author_sort Yang, Chunxiao
collection PubMed
description Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is a reliable technique for quantifying gene expression across various biological processes, of which requires a set of suited reference genes to normalize the expression data. Coleomegilla maculata (Coleoptera: Coccinellidae), is one of the most extensively used biological control agents in the field to manage arthropod pest species. In this study, expression profiles of 16 housekeeping genes selected from C. maculata were cloned and investigated. The performance of these candidates as endogenous controls under specific experimental conditions was evaluated by dedicated algorithms, including geNorm, Normfinder, BestKeeper, and ΔCt method. In addition, RefFinder, a comprehensive platform integrating all the above-mentioned algorithms, ranked the overall stability of these candidate genes. As a result, various sets of suitable reference genes were recommended specifically for experiments involving different tissues, developmental stages, sex, and C. maculate larvae treated with dietary double stranded RNA. This study represents the critical first step to establish a standardized RT-qPCR protocol for the functional genomics research in a ladybeetle C. maculate. Furthermore, it lays the foundation for conducting ecological risk assessment of RNAi-based gene silencing biotechnologies on non-target organisms; in this case, a key predatory biological control agent.
format Online
Article
Text
id pubmed-4674751
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-46747512015-12-16 Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae) Yang, Chunxiao Pan, Huipeng Noland, Jeffrey Edward Zhang, Deyong Zhang, Zhanhong Liu, Yong Zhou, Xuguo Sci Rep Article Reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) is a reliable technique for quantifying gene expression across various biological processes, of which requires a set of suited reference genes to normalize the expression data. Coleomegilla maculata (Coleoptera: Coccinellidae), is one of the most extensively used biological control agents in the field to manage arthropod pest species. In this study, expression profiles of 16 housekeeping genes selected from C. maculata were cloned and investigated. The performance of these candidates as endogenous controls under specific experimental conditions was evaluated by dedicated algorithms, including geNorm, Normfinder, BestKeeper, and ΔCt method. In addition, RefFinder, a comprehensive platform integrating all the above-mentioned algorithms, ranked the overall stability of these candidate genes. As a result, various sets of suitable reference genes were recommended specifically for experiments involving different tissues, developmental stages, sex, and C. maculate larvae treated with dietary double stranded RNA. This study represents the critical first step to establish a standardized RT-qPCR protocol for the functional genomics research in a ladybeetle C. maculate. Furthermore, it lays the foundation for conducting ecological risk assessment of RNAi-based gene silencing biotechnologies on non-target organisms; in this case, a key predatory biological control agent. Nature Publishing Group 2015-12-10 /pmc/articles/PMC4674751/ /pubmed/26656102 http://dx.doi.org/10.1038/srep18201 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Yang, Chunxiao
Pan, Huipeng
Noland, Jeffrey Edward
Zhang, Deyong
Zhang, Zhanhong
Liu, Yong
Zhou, Xuguo
Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae)
title Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae)
title_full Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae)
title_fullStr Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae)
title_full_unstemmed Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae)
title_short Selection of reference genes for RT-qPCR analysis in a predatory biological control agent, Coleomegilla maculata (Coleoptera: Coccinellidae)
title_sort selection of reference genes for rt-qpcr analysis in a predatory biological control agent, coleomegilla maculata (coleoptera: coccinellidae)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4674751/
https://www.ncbi.nlm.nih.gov/pubmed/26656102
http://dx.doi.org/10.1038/srep18201
work_keys_str_mv AT yangchunxiao selectionofreferencegenesforrtqpcranalysisinapredatorybiologicalcontrolagentcoleomegillamaculatacoleopteracoccinellidae
AT panhuipeng selectionofreferencegenesforrtqpcranalysisinapredatorybiologicalcontrolagentcoleomegillamaculatacoleopteracoccinellidae
AT nolandjeffreyedward selectionofreferencegenesforrtqpcranalysisinapredatorybiologicalcontrolagentcoleomegillamaculatacoleopteracoccinellidae
AT zhangdeyong selectionofreferencegenesforrtqpcranalysisinapredatorybiologicalcontrolagentcoleomegillamaculatacoleopteracoccinellidae
AT zhangzhanhong selectionofreferencegenesforrtqpcranalysisinapredatorybiologicalcontrolagentcoleomegillamaculatacoleopteracoccinellidae
AT liuyong selectionofreferencegenesforrtqpcranalysisinapredatorybiologicalcontrolagentcoleomegillamaculatacoleopteracoccinellidae
AT zhouxuguo selectionofreferencegenesforrtqpcranalysisinapredatorybiologicalcontrolagentcoleomegillamaculatacoleopteracoccinellidae