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Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum

A strict relationship exists between the Sirex noctilio and the Amylostereum areolatum, which is carried and spread by its partner. The growth and development of this symbiotic fungus is key to complete the life history of the Sirex woodwasp. Real-time quantitative polymerase chain reaction (RT-qPCR...

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Autores principales: Fu, Ningning, Li, Jiaxing, Wang, Ming, Ren, Lili, Zong, Shixiang, Luo, Youqing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8790227/
https://www.ncbi.nlm.nih.gov/pubmed/35095826
http://dx.doi.org/10.3389/fmicb.2021.827241
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author Fu, Ningning
Li, Jiaxing
Wang, Ming
Ren, Lili
Zong, Shixiang
Luo, Youqing
author_facet Fu, Ningning
Li, Jiaxing
Wang, Ming
Ren, Lili
Zong, Shixiang
Luo, Youqing
author_sort Fu, Ningning
collection PubMed
description A strict relationship exists between the Sirex noctilio and the Amylostereum areolatum, which is carried and spread by its partner. The growth and development of this symbiotic fungus is key to complete the life history of the Sirex woodwasp. Real-time quantitative polymerase chain reaction (RT-qPCR) is used to measure gene expression in samples of A. areolatum at different growth stages and explore the key genes and pathways involved in the growth and development of this symbiotic fungus. To obtain accurate RT-qPCR data, target genes need to be normalized by reference genes that are stably expressed under specific experimental conditions. In our study, the stability of 10 candidate reference genes in symbiotic fungal samples at different growth and development stages was evaluated using geNorm, NormFinder, BestKeeper, delta Ct methods, and RefFinder. Meanwhile, laccase1 was used to validate the stability of the selected reference gene. Under the experimental conditions of this study, p450, CYP, and γ-TUB were identified as suitable reference genes. This work is the first to systematically evaluate the reference genes for RT-qPCR results normalization during the growth of this symbiotic fungus, which lays a foundation for further gene expression experiments and understanding the symbiotic relationship and mechanism between S. noctilio and A. areolatum.
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spelling pubmed-87902272022-01-27 Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum Fu, Ningning Li, Jiaxing Wang, Ming Ren, Lili Zong, Shixiang Luo, Youqing Front Microbiol Microbiology A strict relationship exists between the Sirex noctilio and the Amylostereum areolatum, which is carried and spread by its partner. The growth and development of this symbiotic fungus is key to complete the life history of the Sirex woodwasp. Real-time quantitative polymerase chain reaction (RT-qPCR) is used to measure gene expression in samples of A. areolatum at different growth stages and explore the key genes and pathways involved in the growth and development of this symbiotic fungus. To obtain accurate RT-qPCR data, target genes need to be normalized by reference genes that are stably expressed under specific experimental conditions. In our study, the stability of 10 candidate reference genes in symbiotic fungal samples at different growth and development stages was evaluated using geNorm, NormFinder, BestKeeper, delta Ct methods, and RefFinder. Meanwhile, laccase1 was used to validate the stability of the selected reference gene. Under the experimental conditions of this study, p450, CYP, and γ-TUB were identified as suitable reference genes. This work is the first to systematically evaluate the reference genes for RT-qPCR results normalization during the growth of this symbiotic fungus, which lays a foundation for further gene expression experiments and understanding the symbiotic relationship and mechanism between S. noctilio and A. areolatum. Frontiers Media S.A. 2022-01-12 /pmc/articles/PMC8790227/ /pubmed/35095826 http://dx.doi.org/10.3389/fmicb.2021.827241 Text en Copyright © 2022 Fu, Li, Wang, Ren, Zong and Luo. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Fu, Ningning
Li, Jiaxing
Wang, Ming
Ren, Lili
Zong, Shixiang
Luo, Youqing
Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum
title Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum
title_full Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum
title_fullStr Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum
title_full_unstemmed Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum
title_short Identification and Validation of Reference Genes for Gene Expression Analysis in Different Development Stages of Amylostereum areolatum
title_sort identification and validation of reference genes for gene expression analysis in different development stages of amylostereum areolatum
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8790227/
https://www.ncbi.nlm.nih.gov/pubmed/35095826
http://dx.doi.org/10.3389/fmicb.2021.827241
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