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A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process

Brassinosteroids (BRs) are plant steroid hormones, regulating a broad range of physiological processes. The largest amount of data related with BR biosynthesis has been gathered in Arabidopsis thaliana, however understanding of this process is far less elucidated in monocot crops. Up to now, only fo...

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Autores principales: Gruszka, Damian, Gorniak, Malgorzata, Glodowska, Ewelina, Wierus, Ewa, Oklestkova, Jana, Janeczko, Anna, Maluszynski, Miroslaw, Szarejko, Iwona
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4849053/
https://www.ncbi.nlm.nih.gov/pubmed/27110778
http://dx.doi.org/10.3390/ijms17040600
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author Gruszka, Damian
Gorniak, Malgorzata
Glodowska, Ewelina
Wierus, Ewa
Oklestkova, Jana
Janeczko, Anna
Maluszynski, Miroslaw
Szarejko, Iwona
author_facet Gruszka, Damian
Gorniak, Malgorzata
Glodowska, Ewelina
Wierus, Ewa
Oklestkova, Jana
Janeczko, Anna
Maluszynski, Miroslaw
Szarejko, Iwona
author_sort Gruszka, Damian
collection PubMed
description Brassinosteroids (BRs) are plant steroid hormones, regulating a broad range of physiological processes. The largest amount of data related with BR biosynthesis has been gathered in Arabidopsis thaliana, however understanding of this process is far less elucidated in monocot crops. Up to now, only four barley genes implicated in BR biosynthesis have been identified. Two of them, HvDWARF and HvBRD, encode BR-6-oxidases catalyzing biosynthesis of castasterone, but their relation is not yet understood. In the present study, the identification of the HvDWARF genomic sequence, its mutational and functional analysis and characterization of new mutants are reported. Various types of mutations located in different positions within functional domains were identified and characterized. Analysis of their impact on phenotype of the mutants was performed. The identified homozygous mutants show reduced height of various degree and disrupted skotomorphogenesis. Mutational analysis of the HvDWARF gene with the “reverse genetics” approach allowed for its detailed functional analysis at the level of protein functional domains. The HvDWARF gene function and mutants’ phenotypes were also validated by measurement of endogenous BR concentration. These results allowed a new insight into the BR biosynthesis in barley.
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spelling pubmed-48490532016-05-04 A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process Gruszka, Damian Gorniak, Malgorzata Glodowska, Ewelina Wierus, Ewa Oklestkova, Jana Janeczko, Anna Maluszynski, Miroslaw Szarejko, Iwona Int J Mol Sci Article Brassinosteroids (BRs) are plant steroid hormones, regulating a broad range of physiological processes. The largest amount of data related with BR biosynthesis has been gathered in Arabidopsis thaliana, however understanding of this process is far less elucidated in monocot crops. Up to now, only four barley genes implicated in BR biosynthesis have been identified. Two of them, HvDWARF and HvBRD, encode BR-6-oxidases catalyzing biosynthesis of castasterone, but their relation is not yet understood. In the present study, the identification of the HvDWARF genomic sequence, its mutational and functional analysis and characterization of new mutants are reported. Various types of mutations located in different positions within functional domains were identified and characterized. Analysis of their impact on phenotype of the mutants was performed. The identified homozygous mutants show reduced height of various degree and disrupted skotomorphogenesis. Mutational analysis of the HvDWARF gene with the “reverse genetics” approach allowed for its detailed functional analysis at the level of protein functional domains. The HvDWARF gene function and mutants’ phenotypes were also validated by measurement of endogenous BR concentration. These results allowed a new insight into the BR biosynthesis in barley. MDPI 2016-04-22 /pmc/articles/PMC4849053/ /pubmed/27110778 http://dx.doi.org/10.3390/ijms17040600 Text en © 2016 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gruszka, Damian
Gorniak, Malgorzata
Glodowska, Ewelina
Wierus, Ewa
Oklestkova, Jana
Janeczko, Anna
Maluszynski, Miroslaw
Szarejko, Iwona
A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process
title A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process
title_full A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process
title_fullStr A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process
title_full_unstemmed A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process
title_short A Reverse-Genetics Mutational Analysis of the Barley HvDWARF Gene Results in Identification of a Series of Alleles and Mutants with Short Stature of Various Degree and Disturbance in BR Biosynthesis Allowing a New Insight into the Process
title_sort reverse-genetics mutational analysis of the barley hvdwarf gene results in identification of a series of alleles and mutants with short stature of various degree and disturbance in br biosynthesis allowing a new insight into the process
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4849053/
https://www.ncbi.nlm.nih.gov/pubmed/27110778
http://dx.doi.org/10.3390/ijms17040600
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