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Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla
The northern root-knot nematode (Meloidogyne hapla) is a critical pathogen with a wide host range. Quantitative real-time polymerase chain reaction (qRT-PCR) has been used to elucidate gene expression and function of M. hapla. Suitable reference genes are required to ensure accurate results of qRT-P...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6583968/ https://www.ncbi.nlm.nih.gov/pubmed/31216347 http://dx.doi.org/10.1371/journal.pone.0218610 |
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author | Wu, Xiaojing Yu, Hongyan Yang, Rouwei Zhou, Yuanyuan Zhu, Xiaofeng Wang, Yuanyuan Liu, Xiaoyu Fan, Haiyan Chen, Lijie Duan, Yuxi |
author_facet | Wu, Xiaojing Yu, Hongyan Yang, Rouwei Zhou, Yuanyuan Zhu, Xiaofeng Wang, Yuanyuan Liu, Xiaoyu Fan, Haiyan Chen, Lijie Duan, Yuxi |
author_sort | Wu, Xiaojing |
collection | PubMed |
description | The northern root-knot nematode (Meloidogyne hapla) is a critical pathogen with a wide host range. Quantitative real-time polymerase chain reaction (qRT-PCR) has been used to elucidate gene expression and function of M. hapla. Suitable reference genes are required to ensure accurate results of qRT-PCR for normalising gene expression. Eleven candidate reference genes of M. hapla were selected to evaluate gene expression stability under different conditions. The stability of candidate reference genes was ranked using RefFinder analysis, and the optimal number of reference genes was recommended with geNorm. Notably, the most stable reference genes were SDHA, Mdh, and RpS6 for all samples; SDHA and RpS6 were particularly stable during development stage treatments, whereas Mdh and RpS6 were appropriate for temperature and inorganic compound treatments. In contrast, the least stable reference genes were Actin1 during development stages and all other treatments, GAPDH for temperature treatments, and α-Tub for inorganic compound treatments. One target gene, Mh-Hsp90, was used to verify the selection of reference genes, results showed Mdh and RpS6 could be used as suitable reference genes for M. hapla, and Mdh plus RpS6 were better. Our finding contributes to further work on gene transcription analysis in M. hapla. |
format | Online Article Text |
id | pubmed-6583968 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-65839682019-06-28 Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla Wu, Xiaojing Yu, Hongyan Yang, Rouwei Zhou, Yuanyuan Zhu, Xiaofeng Wang, Yuanyuan Liu, Xiaoyu Fan, Haiyan Chen, Lijie Duan, Yuxi PLoS One Research Article The northern root-knot nematode (Meloidogyne hapla) is a critical pathogen with a wide host range. Quantitative real-time polymerase chain reaction (qRT-PCR) has been used to elucidate gene expression and function of M. hapla. Suitable reference genes are required to ensure accurate results of qRT-PCR for normalising gene expression. Eleven candidate reference genes of M. hapla were selected to evaluate gene expression stability under different conditions. The stability of candidate reference genes was ranked using RefFinder analysis, and the optimal number of reference genes was recommended with geNorm. Notably, the most stable reference genes were SDHA, Mdh, and RpS6 for all samples; SDHA and RpS6 were particularly stable during development stage treatments, whereas Mdh and RpS6 were appropriate for temperature and inorganic compound treatments. In contrast, the least stable reference genes were Actin1 during development stages and all other treatments, GAPDH for temperature treatments, and α-Tub for inorganic compound treatments. One target gene, Mh-Hsp90, was used to verify the selection of reference genes, results showed Mdh and RpS6 could be used as suitable reference genes for M. hapla, and Mdh plus RpS6 were better. Our finding contributes to further work on gene transcription analysis in M. hapla. Public Library of Science 2019-06-19 /pmc/articles/PMC6583968/ /pubmed/31216347 http://dx.doi.org/10.1371/journal.pone.0218610 Text en © 2019 Wu et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Wu, Xiaojing Yu, Hongyan Yang, Rouwei Zhou, Yuanyuan Zhu, Xiaofeng Wang, Yuanyuan Liu, Xiaoyu Fan, Haiyan Chen, Lijie Duan, Yuxi Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla |
title | Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla |
title_full | Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla |
title_fullStr | Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla |
title_full_unstemmed | Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla |
title_short | Evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, Meloidogyne hapla |
title_sort | evaluation of suitable reference genes for gene expression analysis in the northern root-knot nematode, meloidogyne hapla |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6583968/ https://www.ncbi.nlm.nih.gov/pubmed/31216347 http://dx.doi.org/10.1371/journal.pone.0218610 |
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