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...
Autores principales: | , , , , , , |
---|---|
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 |