Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis

Abstract Bromus sterilis is an annual weedy grass, causing high yield losses in winter cereals. Frequent use of herbicides had led to the evolution of herbicide resistance in this species. Mechanisms underlying herbicide resistance in B. sterilis must be uncovered because this problem is becoming a...

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Autores principales:	Madhab Kumar Sen, Kateřina Hamouzová, Pavlina Košnarová, Amit Roy, Josef Soukup
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/221951f3b4bc497facb2de462851c1c1
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spelling	oai:doaj.org-article:221951f3b4bc497facb2de462851c1c12021-12-02T16:10:38ZIdentification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis10.1038/s41598-021-92780-12045-2322https://doaj.org/article/221951f3b4bc497facb2de462851c1c12021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92780-1https://doaj.org/toc/2045-2322Abstract Bromus sterilis is an annual weedy grass, causing high yield losses in winter cereals. Frequent use of herbicides had led to the evolution of herbicide resistance in this species. Mechanisms underlying herbicide resistance in B. sterilis must be uncovered because this problem is becoming a global threat. qRT-PCR and the next-generation sequencing technologies can elucidate the resistance mechanisms. Although qRT-PCR can calculate precise fold changes, its preciseness depends on the expression of reference genes. Regardless of stable expression in any given condition, no gene can act as a universal reference gene. Hence, it is necessary to identify the suitable reference gene for each species. To our knowledge, there are no reports on the suitable reference gene in any brome species so far. Thus, in this paper, the stability of eight genes was evaluated using qRT-PCR experiments followed by expression stability ranking via five most commonly used software for reference gene selection. Our findings suggest using a combination of 18S rRNA and ACCase to normalise the qRT-PCR data in B. sterilis. Besides, reference genes are also recommended for different experimental conditions. The present study outcomes will facilitate future molecular work in B. sterilis and other related grass species.Madhab Kumar SenKateřina HamouzováPavlina KošnarováAmit RoyJosef SoukupNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Madhab Kumar Sen Kateřina Hamouzová Pavlina Košnarová Amit Roy Josef Soukup Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis
description	Abstract Bromus sterilis is an annual weedy grass, causing high yield losses in winter cereals. Frequent use of herbicides had led to the evolution of herbicide resistance in this species. Mechanisms underlying herbicide resistance in B. sterilis must be uncovered because this problem is becoming a global threat. qRT-PCR and the next-generation sequencing technologies can elucidate the resistance mechanisms. Although qRT-PCR can calculate precise fold changes, its preciseness depends on the expression of reference genes. Regardless of stable expression in any given condition, no gene can act as a universal reference gene. Hence, it is necessary to identify the suitable reference gene for each species. To our knowledge, there are no reports on the suitable reference gene in any brome species so far. Thus, in this paper, the stability of eight genes was evaluated using qRT-PCR experiments followed by expression stability ranking via five most commonly used software for reference gene selection. Our findings suggest using a combination of 18S rRNA and ACCase to normalise the qRT-PCR data in B. sterilis. Besides, reference genes are also recommended for different experimental conditions. The present study outcomes will facilitate future molecular work in B. sterilis and other related grass species.
format	article
author	Madhab Kumar Sen Kateřina Hamouzová Pavlina Košnarová Amit Roy Josef Soukup
author_facet	Madhab Kumar Sen Kateřina Hamouzová Pavlina Košnarová Amit Roy Josef Soukup
author_sort	Madhab Kumar Sen
title	Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis
title_short	Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis
title_full	Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis
title_fullStr	Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis
title_full_unstemmed	Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis
title_sort	identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in bromus sterilis
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/221951f3b4bc497facb2de462851c1c1
work_keys_str_mv	AT madhabkumarsen identificationofthemostsuitablereferencegeneforgeneexpressionstudieswithdevelopmentandabioticstressresponseinbromussterilis AT katerinahamouzova identificationofthemostsuitablereferencegeneforgeneexpressionstudieswithdevelopmentandabioticstressresponseinbromussterilis AT pavlinakosnarova identificationofthemostsuitablereferencegeneforgeneexpressionstudieswithdevelopmentandabioticstressresponseinbromussterilis AT amitroy identificationofthemostsuitablereferencegeneforgeneexpressionstudieswithdevelopmentandabioticstressresponseinbromussterilis AT josefsoukup identificationofthemostsuitablereferencegeneforgeneexpressionstudieswithdevelopmentandabioticstressresponseinbromussterilis
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Identification of the most suitable reference gene for gene expression studies with development and abiotic stress response in Bromus sterilis

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