An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background

Plant breeding relies on the meiotic recombination or crossing over to generate the new combinations of the alleles along and among the chromosomes. However, crossing over is constrained in the crops such as barley by a combination of the low frequency and biased distribution. In this study, we atte...

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Autores principales:	Mikel Arrieta, Malcolm Macaulay, Isabelle Colas, Miriam Schreiber, Paul D. Shaw, Robbie Waugh, Luke Ramsay
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	meiosis suppressor screen recombination barley desynaptic mutant crossover Plant culture SB1-1110
Acceso en línea:	https://doaj.org/article/bddb56ed4f4d438aa5222b998c4732da
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spelling	oai:doaj.org-article:bddb56ed4f4d438aa5222b998c4732da2021-11-12T07:05:39ZAn Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background1664-462X10.3389/fpls.2021.706560https://doaj.org/article/bddb56ed4f4d438aa5222b998c4732da2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fpls.2021.706560/fullhttps://doaj.org/toc/1664-462XPlant breeding relies on the meiotic recombination or crossing over to generate the new combinations of the alleles along and among the chromosomes. However, crossing over is constrained in the crops such as barley by a combination of the low frequency and biased distribution. In this study, we attempted to identify the genes that limit the recombination by performing a suppressor screen for the restoration of fertility to the semi-fertile barley mutant desynaptic10 (des10), carrying a mutation in the barley ortholog of MutL-Homolog 3 (HvMLH3), a member of the MutL-homolog (MLH) family of DNA mismatch repair genes. des10 mutants exhibit reduced recombination and fewer chiasmata, resulting in the loss of obligate crossovers (COs) leading to chromosome mis-segregation. We identified several candidate suppressor lines and confirmed their restored fertility in an Hvmlh3 background in the subsequent generations. We focus on one of the candidate suppressor lines, SuppLine2099, which showed the most complete restoration of fertility. We characterized this line by using a target-sequence enrichment and sequencing (TENSEQ) capture array representing barley orthologs of 46 meiotic genes. We found that SuppLine2099 contained a C/T change in the anti-CO gene RecQ-like helicase 4 (RECQL4) resulting in the substitution of a non-polar glycine to a polar aspartic acid (G700D) amino acid in the conserved helicase domain. Single nucleotide polymorphism (SNP) genotyping of F3 populations revealed a significant increase in the recombination frequency in lines with Hvrecql4 in the Hvmlh3 background that was associated with the restoration of fertility. The genotyping also indicated that there was nearly double the recombination levels in homozygous Hvrecql4 lines compared to the wild type (WT). However, we did not observe any significant change in the distribution of CO events. Our results confirm the anti-CO role of RECQL4 in a large genome cereal and establish the possibility of testing the utility of increasing recombination in the context of traditional crop improvement.Mikel ArrietaMalcolm MacaulayIsabelle ColasMiriam SchreiberPaul D. ShawRobbie WaughRobbie WaughLuke RamsayFrontiers Media S.A.articlemeiosissuppressor screenrecombinationbarleydesynaptic mutantcrossoverPlant cultureSB1-1110ENFrontiers in Plant Science, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	meiosis suppressor screen recombination barley desynaptic mutant crossover Plant culture SB1-1110
spellingShingle	meiosis suppressor screen recombination barley desynaptic mutant crossover Plant culture SB1-1110 Mikel Arrieta Malcolm Macaulay Isabelle Colas Miriam Schreiber Paul D. Shaw Robbie Waugh Robbie Waugh Luke Ramsay An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background
description	Plant breeding relies on the meiotic recombination or crossing over to generate the new combinations of the alleles along and among the chromosomes. However, crossing over is constrained in the crops such as barley by a combination of the low frequency and biased distribution. In this study, we attempted to identify the genes that limit the recombination by performing a suppressor screen for the restoration of fertility to the semi-fertile barley mutant desynaptic10 (des10), carrying a mutation in the barley ortholog of MutL-Homolog 3 (HvMLH3), a member of the MutL-homolog (MLH) family of DNA mismatch repair genes. des10 mutants exhibit reduced recombination and fewer chiasmata, resulting in the loss of obligate crossovers (COs) leading to chromosome mis-segregation. We identified several candidate suppressor lines and confirmed their restored fertility in an Hvmlh3 background in the subsequent generations. We focus on one of the candidate suppressor lines, SuppLine2099, which showed the most complete restoration of fertility. We characterized this line by using a target-sequence enrichment and sequencing (TENSEQ) capture array representing barley orthologs of 46 meiotic genes. We found that SuppLine2099 contained a C/T change in the anti-CO gene RecQ-like helicase 4 (RECQL4) resulting in the substitution of a non-polar glycine to a polar aspartic acid (G700D) amino acid in the conserved helicase domain. Single nucleotide polymorphism (SNP) genotyping of F3 populations revealed a significant increase in the recombination frequency in lines with Hvrecql4 in the Hvmlh3 background that was associated with the restoration of fertility. The genotyping also indicated that there was nearly double the recombination levels in homozygous Hvrecql4 lines compared to the wild type (WT). However, we did not observe any significant change in the distribution of CO events. Our results confirm the anti-CO role of RECQL4 in a large genome cereal and establish the possibility of testing the utility of increasing recombination in the context of traditional crop improvement.
format	article
author	Mikel Arrieta Malcolm Macaulay Isabelle Colas Miriam Schreiber Paul D. Shaw Robbie Waugh Robbie Waugh Luke Ramsay
author_facet	Mikel Arrieta Malcolm Macaulay Isabelle Colas Miriam Schreiber Paul D. Shaw Robbie Waugh Robbie Waugh Luke Ramsay
author_sort	Mikel Arrieta
title	An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background
title_short	An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background
title_full	An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background
title_fullStr	An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background
title_full_unstemmed	An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background
title_sort	induced mutation in hvrecql4 increases the overall recombination and restores fertility in a barley hvmlh3 mutant background
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/bddb56ed4f4d438aa5222b998c4732da
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An Induced Mutation in HvRECQL4 Increases the Overall Recombination and Restores Fertility in a Barley HvMLH3 Mutant Background

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