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Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)

Buffalobur (Solanum rostratum Dunal), which belongs to the Solanaceae family, is a worldwide noxious invasive weed and is listed as one of the top 10 alien invasive species in China. It is harmful to humans and livestock because the entire plant is covered with spines containing toxins. Many studies...

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Autores principales: Zhao, Dandan, Wang, Xu, Chen, Jingchao, Huang, Zhaofeng, Huo, Heqiang, Jiang, Cuilan, Huang, Hongjuan, Zhang, Chaoxian, Wei, Shouhui
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6502881/
https://www.ncbi.nlm.nih.gov/pubmed/31061419
http://dx.doi.org/10.1038/s41598-019-43438-6
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author Zhao, Dandan
Wang, Xu
Chen, Jingchao
Huang, Zhaofeng
Huo, Heqiang
Jiang, Cuilan
Huang, Hongjuan
Zhang, Chaoxian
Wei, Shouhui
author_facet Zhao, Dandan
Wang, Xu
Chen, Jingchao
Huang, Zhaofeng
Huo, Heqiang
Jiang, Cuilan
Huang, Hongjuan
Zhang, Chaoxian
Wei, Shouhui
author_sort Zhao, Dandan
collection PubMed
description Buffalobur (Solanum rostratum Dunal), which belongs to the Solanaceae family, is a worldwide noxious invasive weed and is listed as one of the top 10 alien invasive species in China. It is harmful to humans and livestock because the entire plant is covered with spines containing toxins. Many studies have analysed the gene expression in this weed species under different stress conditions using quantitative real-time PCR (qPCR). However, until now, there has been no report on suitable reference genes in buffalobur. Herein, 14 candidate reference genes were selected and evaluated for their expression stability in buffalobur in different tissues, at different developmental stages, and in response to several stress conditions using the geNorm, NormFinder, BestKeeper and RefFinder statistical algorithms. The results showed that EF1α, ACT and SAND are suitable reference genes across all samples tested. We recommend the normalization of target gene expression under different experimental conditions using these three genes together. Validation of selected reference genes was achieved by assessing the relative expression levels of P5CS and GI. This work identified the appropriate reference genes for transcript normalization in buffalobur, which will be helpful in future genetic studies of this invasive weed.
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spelling pubmed-65028812019-05-20 Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal) Zhao, Dandan Wang, Xu Chen, Jingchao Huang, Zhaofeng Huo, Heqiang Jiang, Cuilan Huang, Hongjuan Zhang, Chaoxian Wei, Shouhui Sci Rep Article Buffalobur (Solanum rostratum Dunal), which belongs to the Solanaceae family, is a worldwide noxious invasive weed and is listed as one of the top 10 alien invasive species in China. It is harmful to humans and livestock because the entire plant is covered with spines containing toxins. Many studies have analysed the gene expression in this weed species under different stress conditions using quantitative real-time PCR (qPCR). However, until now, there has been no report on suitable reference genes in buffalobur. Herein, 14 candidate reference genes were selected and evaluated for their expression stability in buffalobur in different tissues, at different developmental stages, and in response to several stress conditions using the geNorm, NormFinder, BestKeeper and RefFinder statistical algorithms. The results showed that EF1α, ACT and SAND are suitable reference genes across all samples tested. We recommend the normalization of target gene expression under different experimental conditions using these three genes together. Validation of selected reference genes was achieved by assessing the relative expression levels of P5CS and GI. This work identified the appropriate reference genes for transcript normalization in buffalobur, which will be helpful in future genetic studies of this invasive weed. Nature Publishing Group UK 2019-05-06 /pmc/articles/PMC6502881/ /pubmed/31061419 http://dx.doi.org/10.1038/s41598-019-43438-6 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Zhao, Dandan
Wang, Xu
Chen, Jingchao
Huang, Zhaofeng
Huo, Heqiang
Jiang, Cuilan
Huang, Hongjuan
Zhang, Chaoxian
Wei, Shouhui
Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
title Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
title_full Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
title_fullStr Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
title_full_unstemmed Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
title_short Selection of reference genes for qPCR normalization in buffalobur (Solanum rostratum Dunal)
title_sort selection of reference genes for qpcr normalization in buffalobur (solanum rostratum dunal)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6502881/
https://www.ncbi.nlm.nih.gov/pubmed/31061419
http://dx.doi.org/10.1038/s41598-019-43438-6
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