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1.
Plant J ; 119(1): 364-382, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38652034

RESUMEN

Barley produces several specialized metabolites, including five α-, ß-, and γ-hydroxynitrile glucosides (HNGs). In malting barley, presence of the α-HNG epiheterodendrin gives rise to undesired formation of ethyl carbamate in the beverage production, especially after distilling. Metabolite-GWAS identified QTLs and underlying gene candidates possibly involved in the control of the relative and absolute content of HNGs, including an undescribed MATE transporter. By screening 325 genetically diverse barley accessions, we discovered three H. vulgare ssp. spontaneum (wild barley) lines with drastic changes in the relative ratios of the five HNGs. Knock-out (KO)-lines, isolated from the barley FIND-IT resource and each lacking one of the functional HNG biosynthetic genes (CYP79A12, CYP71C103, CYP71C113, CYP71U5, UGT85F22 and UGT85F23) showed unprecedented changes in HNG ratios enabling assignment of specific and mutually dependent catalytic functions to the biosynthetic enzymes involved. The highly similar relative ratios between the five HNGs found across wild and domesticated barley accessions indicate assembly of the HNG biosynthetic enzymes in a metabolon, the functional output of which was reconfigured in the absence of a single protein component. The absence or altered ratios of the five HNGs in the KO-lines did not change susceptibility to the fungal phytopathogen Pyrenophora teres causing net blotch. The study provides a deeper understanding of the organization of HNG biosynthesis in barley and identifies a novel, single gene HNG-0 line in an elite spring barley background for direct use in breeding of malting barley, eliminating HNGs as a source of ethyl carbamate formation in whisky production.


Asunto(s)
Glucósidos , Hordeum , Hordeum/genética , Hordeum/metabolismo , Hordeum/microbiología , Glucósidos/metabolismo , Nitrilos/metabolismo , Sitios de Carácter Cuantitativo , Uretano/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estudio de Asociación del Genoma Completo
2.
Development ; 148(5)2021 03 09.
Artículo en Inglés | MEDLINE | ID: mdl-33526582

RESUMEN

Cereal grain develops from fertilised florets. Alterations in floret and grain development greatly influence grain yield and quality. Despite this, little is known about the underlying genetic control of these processes, especially in key temperate cereals such as barley and wheat. Using a combination of near-isogenic mutant comparisons, gene editing and genetic analyses, we reveal that HvAPETALA2 (HvAP2) controls floret organ identity, floret boundaries, and maternal tissue differentiation and elimination during grain development. These new roles of HvAP2 correlate with changes in grain size and HvAP2-dependent expression of specific HvMADS-box genes, including the B-sister gene, HvMADS29 Consistent with this, gene editing demonstrates that HvMADS29 shares roles with HvAP2 in maternal tissue differentiation. We also discovered that a gain-of-function HvAP2 allele masks changes in floret organ identity and grain size due to loss of barley LAXATUM.A/BLADE-ON-PETIOLE2 (HvBOP2) gene function. Taken together, we reveal novel pleiotropic roles and regulatory interactions for an AP2-like gene controlling floret and grain development in a temperate cereal.


Asunto(s)
Proteínas de Homeodominio/metabolismo , Hordeum/metabolismo , Proteínas de Dominio MADS/metabolismo , Proteínas de Plantas/metabolismo , Alelos , Secuencia de Bases , Sistemas CRISPR-Cas/genética , Grano Comestible/anatomía & histología , Grano Comestible/metabolismo , Flores/crecimiento & desarrollo , Flores/metabolismo , Edición Génica , Regulación de la Expresión Génica de las Plantas , Genotipo , Proteínas de Homeodominio/química , Proteínas de Homeodominio/genética , Hordeum/crecimiento & desarrollo , Proteínas de Dominio MADS/genética , Mutagénesis , Fenotipo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
3.
New Phytol ; 237(6): 2136-2147, 2023 03.
Artículo en Inglés | MEDLINE | ID: mdl-36600397

RESUMEN

In cereal species, grain size is influenced by growth of the ovule integuments (seed coat), the spikelet hull (lemma and palea) and the filial endosperm. Whether a highly conserved ovule tissue, the nucellus, has any impact on grain size has remained unclear. Immunolabelling revealed that the barley nucellus comprises two distinct cell types that differ in terms of cell wall homogalacturonan (HG) accumulation. Transcriptional profiling of the nucellus identified two pectin methylesterase (PME) genes, OVULE PECTIN MODIFIER 1 (OPM1) and OPM2, which are expressed in the unfertilized ovule but absent from the seed. Ovules from an opm1 opm2 mutant and plants expressing an ovule-specific pectin methylesterase inhibitor (PMEI), exhibit reduced HG accumulation. This results in changes to ovule cell size and shape and ovules that are longer than wild-type (WT) controls. At grain maturity, this is manifested as significantly longer grain. These findings indicate that cell wall composition during ovule development acts to limit ovule and seed growth. The investigation of ovule PME and PMEI activity reveals an unexpected role of maternal tissues in controlling grain growth before fertilization, one that has been lacking from models exploring improvements in grain size.


Asunto(s)
Grano Comestible , Hordeum , Grano Comestible/genética , Óvulo Vegetal/metabolismo , Hordeum/genética , Semillas/genética , Pared Celular , Regulación de la Expresión Génica de las Plantas
4.
Plant J ; 108(2): 509-527, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34382710

RESUMEN

Transition to the reproductive phase, inflorescence formation and flower development are crucial elements that ensure maximum reproductive success in a plant's life cycle. To understand the regulatory mechanisms underlying correct flower development in barley (Hordeum vulgare), we characterized the multiovary 5 (mov5.o) mutant. This mutant develops abnormal flowers that exhibit mosaic floral organs typified by multiple carpels at the total or partial expense of stamens. Genetic mapping positioned mov5 on the long arm of chromosome 2H, incorporating a region that encodes HvLFY, the barley orthologue of LEAFY from Arabidopsis. Sequencing revealed that, in mov5.o plants, HvLFY contains a single amino acid substitution in a highly conserved proline residue. CRISPR-mediated knockout of HvLFY replicated the mov5.o phenotype, suggesting that HvLFYmov5 represents a loss of function allele. In heterologous assays, the HvLFYmov5 polymorphism influenced protein-protein interactions and affinity for a putative binding site in the promoter of HvMADS58, a C-class MADS-box gene. Moreover, molecular analysis indicated that HvLFY interacts with HvUFO and regulates the expression of floral homeotic genes including HvMADS2, HvMADS4 and HvMADS16. Other distinct changes in expression differ from those reported in the rice LFY mutants apo2/rfl, suggesting that LFY function in the grasses is modulated in a species-specific manner. This pathway provides a key entry point for the study of LFY function and multiple ovary formation in barley, as well as cereal species in general.


Asunto(s)
Flores/crecimiento & desarrollo , Hordeum/fisiología , Proteínas de Plantas/genética , Sustitución de Aminoácidos , Proteínas de Arabidopsis/genética , Sitios de Unión , Mapeo Cromosómico , Cromosomas de las Plantas , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , ADN de Plantas/metabolismo , Flores/fisiología , Regulación de la Expresión Génica de las Plantas , Genes Homeobox , Hordeum/crecimiento & desarrollo , Inflorescencia/genética , Mutación , Proteínas de Plantas/metabolismo , Polimorfismo de Nucleótido Simple , Factores de Transcripción/genética
5.
Plant J ; 104(4): 1009-1022, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-32890421

RESUMEN

Barley (Hordeum vulgare L) grain is comparatively rich in (1,3;1,4)-ß-glucan, a source of fermentable dietary fibre that protects against various human health conditions. However, low grain (1,3;1,4)-ß-glucan content is preferred for brewing and distilling. We took a reverse genetics approach, using CRISPR/Cas9 to generate mutations in members of the Cellulose synthase-like (Csl) gene superfamily that encode known (HvCslF6 and HvCslH1) and putative (HvCslF3 and HvCslF9) (1,3;1,4)-ß-glucan synthases. Resultant mutations ranged from single amino acid (aa) substitutions to frameshift mutations causing premature stop codons, and led to specific differences in grain morphology, composition and (1,3;1,4)-ß-glucan content. (1,3;1,4)-ß-Glucan was absent in the grain of cslf6 knockout lines, whereas cslf9 knockout lines had similar (1,3;1,4)-ß-glucan content to wild-type (WT). However, cslf9 mutants showed changes in the abundance of other cell-wall-related monosaccharides compared with WT. Thousand grain weight (TGW), grain length, width and surface area were altered in cslf6 knockouts, and to a lesser extent TGW in cslf9 knockouts. cslf3 and cslh1 mutants had no effect on grain (1,3;1,4)-ß-glucan content. Our data indicate that multiple members of the CslF/H family fulfil important functions during grain development but, with the exception of HvCslF6, do not impact the abundance of (1,3;1,4)-ß-glucan in mature grain.


Asunto(s)
Hordeum/enzimología , Proteínas de Plantas/metabolismo , beta-Glucanos/metabolismo , Pared Celular/metabolismo , Grano Comestible , Edición Génica , Glucosiltransferasas/genética , Glucosiltransferasas/metabolismo , Hordeum/genética , Mutagénesis Sitio-Dirigida , Mutación , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Polisacáridos/metabolismo
6.
J Exp Bot ; 72(7): 2383-2402, 2021 03 29.
Artículo en Inglés | MEDLINE | ID: mdl-33421064

RESUMEN

We profiled the grain oligosaccharide content of 154 two-row spring barley genotypes and quantified 27 compounds, mainly inulin- and neoseries-type fructans, showing differential abundance. Clustering revealed two profile groups where the 'high' set contained greater amounts of sugar monomers, sucrose, and overall fructans, but lower fructosylraffinose. A genome-wide association study (GWAS) identified a significant association for the variability of two fructan types: neoseries-DP7 and inulin-DP9, which showed increased strength when applying a novel compound ratio-GWAS approach. Gene models within this region included three known fructan biosynthesis genes (fructan:fructan 1-fructosyltransferase, sucrose:sucrose 1-fructosyltransferase, and sucrose:fructan 6-fructosyltransferase). Two other genes in this region, 6(G)-fructosyltransferase and vacuolar invertase1, have not previously been linked to fructan biosynthesis and showed expression patterns distinct from those of the other three genes, including exclusive expression of 6(G)-fructosyltransferase in outer grain tissues at the storage phase. From exome capture data, several single nucleotide polymorphisms related to inulin- and neoseries-type fructan variability were identified in fructan:fructan 1-fructosyltransferase and 6(G)-fructosyltransferase genes. Co-expression analyses uncovered potential regulators of fructan biosynthesis including transcription factors. Our results provide the first scientific evidence for the distinct biosynthesis of neoseries-type fructans during barley grain maturation and reveal novel gene candidates likely to be involved in the differential biosynthesis of various types of fructan in barley.


Asunto(s)
Hexosiltransferasas , Hordeum , Secuencia de Aminoácidos , Fructanos , Estudio de Asociación del Genoma Completo , Hexosiltransferasas/genética , Hexosiltransferasas/metabolismo , Hordeum/genética , Hordeum/metabolismo , Vacuolas/metabolismo
7.
Plant Physiol ; 175(4): 1720-1731, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29101279

RESUMEN

Increasing grain yield is an endless challenge for cereal crop breeding. In barley (Hordeum vulgare), grain number is controlled mainly by Six-rowed spike 1 (Vrs1), which encodes a homeodomain leucine zipper class I transcription factor. However, little is known about the genetic basis of grain size. Here, we show that extreme suppression of lateral florets contributes to enlarged grains in deficiens barley. Through a combination of fine-mapping and resequencing of deficiens mutants, we have identified that a single amino acid substitution at a putative phosphorylation site in VRS1 is responsible for the deficiens phenotype. deficiens mutant alleles confer an increase in grain size, a reduction in plant height, and a significant increase in thousand grain weight in contemporary cultivated germplasm. Haplotype analysis revealed that barley carrying the deficiens allele (Vrs1.t1) originated from two-rowed types carrying the Vrs1.b2 allele, predominantly found in germplasm from northern Africa. In situ hybridization of histone H4, a marker for cell cycle or proliferation, showed weaker expression in the lateral spikelets compared with central spikelets in deficiens Transcriptome analysis revealed that a number of histone superfamily genes were up-regulated in the deficiens mutant, suggesting that enhanced cell proliferation in the central spikelet may contribute to larger grains. Our data suggest that grain yield can be improved by suppressing the development of specific organs that are not positively involved in sink/source relationships.


Asunto(s)
Regulación de la Expresión Génica de las Plantas/fisiología , Hordeum/fisiología , Proteínas de Plantas/metabolismo , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Cruzamiento , Mapeo Cromosómico , Cromosomas de las Plantas/genética , Marcadores Genéticos , Estudio de Asociación del Genoma Completo , Haplotipos , Hordeum/genética , Mutación , Desarrollo de la Planta/genética , Desarrollo de la Planta/fisiología , Proteínas de Plantas/química , Proteínas de Plantas/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN de Planta/genética , ARN de Planta/metabolismo , Transcripción Genética
8.
BMC Genomics ; 18(1): 122, 2017 01 31.
Artículo en Inglés | MEDLINE | ID: mdl-28143400

RESUMEN

BACKGROUND: In plants carotenoids play an important role in the photosynthetic process and photo-oxidative protection, and are the substrate for the synthesis of abscisic acid and strigolactones. In addition to their protective role as antioxidants and precursors of vitamin A, in wheat carotenoids are important as they influence the colour (whiteness vs. yellowness) of the grain. Understanding the genetic basis of grain yellow pigments, and identifying associated markers provide the basis for improving wheat quality by molecular breeding. RESULTS: Twenty-four candidate genes involved in the biosynthesis and catabolism of carotenoid compounds have been identified in wheat by comparative genomics. Single nucleotide polymorphisms (SNPs) found in the coding sequences of 19 candidate genes allowed their chromosomal location and accurate map position on two reference consensus maps to be determined. The genome-wide association study based on genotyping a tetraploid wheat collection with 81,587 gene-associated SNPs validated quantitative trait loci (QTLs) previously detected in biparental populations and discovered new QTLs for grain colour-related traits. Ten carotenoid genes mapped in chromosome regions underlying pigment content QTLs indicating possible functional relationships between candidate genes and the trait. CONCLUSIONS: The availability of linked, candidate gene-based markers can facilitate breeding wheat cultivars with desirable levels of carotenoids. Identifying QTLs linked to carotenoid pigmentation can contribute to understanding genes underlying carotenoid accumulation in the wheat kernels. Together these outputs can be combined to exploit the genetic variability of colour-related traits for the nutritional and commercial improvement of wheat products.


Asunto(s)
Carotenoides/metabolismo , Pigmentación/genética , Pigmentos Biológicos/metabolismo , Triticum/genética , Triticum/metabolismo , Carotenoides/biosíntesis , Mapeo Cromosómico , Estudios de Asociación Genética , Estudio de Asociación del Genoma Completo , Redes y Vías Metabólicas , Fenotipo , Filogenia , Polimorfismo de Nucleótido Simple , Sitios de Carácter Cuantitativo , Triticum/clasificación
9.
Plant Physiol ; 170(3): 1549-65, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26754666

RESUMEN

Within the cereal grain, the endosperm and its nutrient reserves are critical for successful germination and in the context of grain utilization. The identification of molecular determinants of early endosperm development, particularly regulators of cell division and cell wall deposition, would help predict end-use properties such as yield, quality, and nutritional value. Custom microarray data have been generated using RNA isolated from developing barley grain endosperm 3 d to 8 d after pollination (DAP). Comparisons of transcript abundance over time revealed 47 gene expression modules that can be clustered into 10 broad groups. Superimposing these modules upon cytological data allowed patterns of transcript abundance to be linked with key stages of early grain development. Here, attention was focused on how the datasets could be mined to explore and define the processes of cell wall biosynthesis, remodeling, and degradation. Using a combination of spatial molecular network and gene ontology enrichment analyses, it is shown that genes involved in cell wall metabolism are found in multiple modules, but cluster into two main groups that exhibit peak expression at 3 DAP to 4 DAP and 5 DAP to 8 DAP. The presence of transcription factor genes in these modules allowed candidate genes for the control of wall metabolism during early barley grain development to be identified. The data are publicly available through a dedicated web interface (https://ics.hutton.ac.uk/barseed/), where they can be used to interrogate co- and differential expression for any other genes, groups of genes, or transcription factors expressed during early endosperm development.


Asunto(s)
Endospermo/genética , Perfilación de la Expresión Génica/métodos , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Hordeum/genética , Pared Celular/genética , Pared Celular/metabolismo , Análisis por Conglomerados , Grano Comestible/citología , Grano Comestible/embriología , Grano Comestible/genética , Endospermo/citología , Endospermo/embriología , Ontología de Genes , Redes Reguladoras de Genes , Hordeum/citología , Hordeum/embriología , Análisis de Secuencia por Matrices de Oligonucleótidos , Proteínas de Plantas/clasificación , Proteínas de Plantas/genética , Polinización/genética , Factores de Tiempo
10.
Proc Natl Acad Sci U S A ; 110(41): 16675-80, 2013 Oct 08.
Artículo en Inglés | MEDLINE | ID: mdl-24065816

RESUMEN

Within the cereal grasses, variation in inflorescence architecture results in a conspicuous morphological diversity that in crop species influences the yield of cereal grains. Although significant progress has been made in identifying some of the genes underlying this variation in maize and rice, in the temperate cereals, a group that includes wheat, barley, and rye, only the dosage-dependent and highly pleiotropic Q locus in hexaploid wheat has been molecularly characterized. Here we show that the characteristic variation in the density of grains along the inflorescence, or spike, of modern cultivated barley (Hordeum vulgare) is largely the consequence of a perturbed interaction between microRNA172 and its corresponding binding site in the mRNA of an APELATA2 (AP2)-like transcription factor, HvAP2. We used genome-wide association and biparental mapping to identify HvAP2. By comparing inflorescence development and HvAP2 transcript abundance in an extreme dense-spike mutant and its nearly isogenic WT line, we show that HvAP2 turnover driven by microRNA 172 regulates the length of a critical developmental window that is required for elongation of the inflorescence internodes. Our data indicate that this heterochronic change, an altered timing of developmental events caused by specific temporal variation in the efficiency of HvAP2 turnover, leads to the striking differences in the size and shape of the barley spike.


Asunto(s)
Flores/fisiología , Hordeum/genética , MicroARNs/metabolismo , Semillas/fisiología , Factores de Transcripción/metabolismo , Secuencia de Bases , Cartilla de ADN/genética , Flores/genética , Flores/ultraestructura , Estudio de Asociación del Genoma Completo , Hordeum/fisiología , MicroARNs/genética , Microscopía Electrónica de Rastreo , Datos de Secuencia Molecular , Análisis de Componente Principal , Reacción en Cadena en Tiempo Real de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADN , Factores de Transcripción/genética
11.
BMC Genomics ; 15: 907, 2014 Oct 17.
Artículo en Inglés | MEDLINE | ID: mdl-25326272

RESUMEN

BACKGROUND: (1,3;1,4)-ß-Glucan is an important component of the cell walls of barley grain as it affects processability during the production of alcoholic beverages and has significant human health benefits when consumed above recommended threshold levels. This leads to diametrically opposed quality requirements for different applications as low levels of (1,3;1,4)-ß-glucan are required for brewing and distilling and high levels for positive impacts on human health. RESULTS: We quantified grain (1,3;1,4)-ß-glucan content in a collection of 399 2-row Spring-type, and 204 2-row Winter-type elite barley cultivars originating mainly from north western Europe. We combined these data with genotypic information derived using a 9 K Illumina iSelect SNP platform and subsequently carried out a Genome Wide Association Scan (GWAS). Statistical analysis accounting for residual genetic structure within the germplasm collection allowed us to identify significant associations between molecular markers and the phenotypic data. By anchoring the regions that contain these associations to the barley genome assembly we catalogued genes underlying the associations. Based on gene annotations and transcript abundance data we identified candidate genes. CONCLUSIONS: We show that a region of the genome on chromosome 2 containing a cluster of CELLULOSE SYNTHASE-LIKE (Csl) genes, including CslF3, CslF4, CslF8, CslF10, CslF12 and CslH, as well as a region on chromosome 1H containing CslF9, are associated with the phenotype in this germplasm. We also observed that several regions identified by GWAS contain glycoside hydrolases that are possibly involved in (1,3;1,4)-ß-glucan breakdown, together with other genes that might participate in (1,3;1,4)-ß-glucan synthesis, re-modelling or regulation. This analysis provides new opportunities for understanding the genes related to the regulation of (1,3;1,4)-ß-glucan content in cereal grains.


Asunto(s)
Genómica , Hordeum/genética , Hordeum/metabolismo , Estaciones del Año , beta-Glucanos/metabolismo , Regulación de la Expresión Génica de las Plantas , Genoma de Planta/genética , Glucosiltransferasas/genética , Glicósido Hidrolasas/genética , Hordeum/enzimología , Hordeum/crecimiento & desarrollo , Polimorfismo de Nucleótido Simple/genética
12.
Front Plant Sci ; 15: 1268847, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38571708

RESUMEN

In the last century, breeding programs have traditionally favoured yield-related traits, grown under high-input conditions, resulting in a loss of genetic diversity and an increased susceptibility to stresses in crops. Thus, exploiting understudied genetic resources, that potentially harbour tolerance genes, is vital for sustainable agriculture. Northern European barley germplasm has been relatively understudied despite its key role within the malting industry. The European Heritage Barley collection (ExHIBiT) was assembled to explore the genetic diversity in European barley focusing on Northern European accessions and further address environmental pressures. ExHIBiT consists of 363 spring-barley accessions, focusing on two-row type. The collection consists of landraces (~14%), old cultivars (~18%), elite cultivars (~67%) and accessions with unknown breeding history (~1%), with 70% of the collection from Northern Europe. The population structure of the ExHIBiT collection was subdivided into three main clusters primarily based on the accession's year of release using 26,585 informative SNPs based on 50k iSelect single nucleotide polymorphism (SNP) array data. Power analysis established a representative core collection of 230 genotypically and phenotypically diverse accessions. The effectiveness of this core collection for conducting statistical and association analysis was explored by undertaking genome-wide association studies (GWAS) using 24,876 SNPs for nine phenotypic traits, four of which were associated with SNPs. Genomic regions overlapping with previously characterised flowering genes (HvZTLb) were identified, demonstrating the utility of the ExHIBiT core collection for locating genetic regions that determine important traits. Overall, the ExHIBiT core collection represents the high level of untapped diversity within Northern European barley, providing a powerful resource for researchers and breeders to address future climate scenarios.

13.
Front Plant Sci ; 14: 1095862, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37235033

RESUMEN

The phenolic acids, ferulic acid and p-coumaric acid, are components of plant cell walls in grasses, including many of our major food crops. They have important health-promoting properties in grain, and influence the digestibility of biomass for industrial processing and livestock feed. Both phenolic acids are assumed to be critical to cell wall integrity and ferulic acid, at least, is important for cross-linking cell wall components, but the role of p-coumaric acid is unclear. Here we identify alleles of a BAHD p-coumaroyl arabinoxylan transferase, HvAT10, as responsible for the natural variation in cell wall-esterified phenolic acids in whole grain within a cultivated two-row spring barley panel. We show that HvAT10 is rendered non-functional by a premature stop codon mutation in half of the genotypes in our mapping panel. This results in a dramatic reduction in grain cell wall-esterifed p-coumaric acid, a moderate rise in ferulic acid, and a clear increase in the ferulic acid to p-coumaric acid ratio. The mutation is virtually absent in wild and landrace germplasm suggesting an important function for grain arabinoxylan p-coumaroylation pre-domestication that is dispensable in modern agriculture. Intriguingly, we detected detrimental impacts of the mutated locus on grain quality traits where it was associated with smaller grain and poorer malting properties. HvAT10 could be a focus for improving grain quality for malting or phenolic acid content in wholegrain foods.

14.
Plant Physiol ; 155(2): 617-27, 2011 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-21088227

RESUMEN

Since the early 20th century, barley (Hordeum vulgare) has been a model for investigating the effects of physical and chemical mutagens and for exploring the potential of mutation breeding in crop improvement. As a consequence, extensive and well-characterized collections of morphological and developmental mutants have been assembled that represent a valuable resource for exploring a wide range of complex and fundamental biological processes. We constructed a collection of 881 backcrossed lines containing mutant alleles that induce a majority of the morphological and developmental variation described in this species. After genotyping these lines with up to 3,072 single nucleotide polymorphisms, comparison to their recurrent parent defined the genetic location of 426 mutant alleles to chromosomal segments, each representing on average <3% of the barley genetic map. We show how the gene content in these segments can be predicted through conservation of synteny with model cereal genomes, providing a route to rapid gene identification.


Asunto(s)
Genómica/métodos , Genotipo , Hordeum/genética , Alelos , Mapeo Cromosómico , Cruzamientos Genéticos , ADN de Plantas/genética , Genes de Plantas , Hordeum/crecimiento & desarrollo , Mutación , Oryza/genética , Polimorfismo de Nucleótido Simple , Sintenía
15.
Theor Appl Genet ; 125(1): 33-45, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22395962

RESUMEN

A typical barley (Hordeum vulgare) floret consists of reproductive organs three stamens and a pistil, and non-reproductive organs-lodicules and two floral bracts, abaxial called 'lemma' and adaxial 'palea'. The floret is subtended by two additional bracts called outer or empty glumes. Together these organs form the basic structural unit of the grass inflorescence, a spikelet. There are commonly three spikelets at each rachis (floral stem of the barley spike) node, one central and two lateral spikelets. Rare naturally occurring or induced phenotypic variants that contain a third bract subtending the central spikelets have been described in barley. The gene responsible for this phenotype was called the THIRD OUTER GLUME1 (Trd1). The Trd1 mutants fail to suppress bract growth and as a result produce leaf-like structures that subtend each rachis node in the basal portion of the spike. Also, floral development at the collar is not always suppressed. In rice and maize, recessive mutations in NECK LEAF1 (Nl1) and TASSEL SHEATH1 (Tsh1) genes, respectively, have been shown to be responsible for orthologous phenotypes. Fine mapping of the trd1 phenotype in an F(3) recombinant population enabled us to position Trd1 on the long arm of chromosome 1H to a 10 cM region. We anchored this to a conserved syntenic region on rice chromosome Os05 and selected a set of candidate genes for validation by resequencing PCR amplicons from a series of independent mutant alleles. This analysis revealed that a GATA transcription factor, recently proposed to be Trd1, contained mutations in 10 out of 14 independent trd1 mutant alleles that would generate non-functional TRD1 proteins. Together with genetic linkage data, we confirm the identity of Trd1 as the GATA transcription factor ortholog of rice Nl1 and maize Tsh1 genes.


Asunto(s)
Flores/genética , Genes de Plantas/genética , Genes Supresores , Hordeum/anatomía & histología , Hordeum/genética , Supresión Genética , Regiones no Traducidas 3'/genética , Regiones no Traducidas 5'/genética , Alelos , Secuencia de Bases , Cromosomas de las Plantas/genética , Cruzamientos Genéticos , Flores/ultraestructura , Estudios de Asociación Genética , Hordeum/ultraestructura , Datos de Secuencia Molecular , Mutación/genética , Oryza/genética , Fenotipo , Mapeo Físico de Cromosoma , Polimorfismo Genético
16.
Front Plant Sci ; 13: 883139, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36160970

RESUMEN

(1,3;1,4)-ß-Glucan is a non-cellulosic polysaccharide required for correct barley grain fill and plant development, with industrial relevance in the brewing and the functional food sector. Barley grains contain higher levels of (1,3;1,4)-ß-glucan compared to other small grain cereals and this influences their end use, having undesirable effects on brewing and distilling and beneficial effects linked to human health. HvCslF6 is the main gene contributing to (1,3;1,4)-ß-glucan biosynthesis in the grain. Here, the transcriptional regulation of HvCslF6 was investigated using an in-silico analysis of transcription factor binding sites (TFBS) in its putative promoter, and functional characterization in a barley protoplast transient expression system. Based on TFBS predictions, TF classes AP2/ERF, MYB, and basic helix-loop-helix (bHLH) were over-represented within a 1,000 bp proximal HvCslF6 promoter region. Dual luciferase assays based on multiple HvCslF6 deletion constructs revealed the promoter fragment driving HvCslF6 expression. Highest HvCslF6 promoter activity was narrowed down to a 51 bp region located -331 bp to -382 bp upstream of the start codon. We combined this with TFBS predictions to identify two MYB TFs: HvMYB61 and HvMYB46/83 as putative activators of HvCslF6 expression. Gene network analyses assigned HvMYB61 to the same co-expression module as HvCslF6 and other primary cellulose synthases (HvCesA1, HvCesA2, and HvCesA6), whereas HvMYB46/83 was assigned to a different module. Based on RNA-seq expression during grain development, HvMYB61 was cloned and tested in the protoplast system. The transient over-expression of HvMYB61 in barley protoplasts suggested a positive regulatory effect on HvCslF6 expression.

17.
Biochim Biophys Acta Biomembr ; 1863(10): 183669, 2021 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-34139196

RESUMEN

Leaf Na+ exclusion, mediated by plasma membrane-localised Class 1 High-affinity potassium (K+) Transporters (HKTs), is a key mechanism contributing to salinity tolerance of several major crop plants. We determined previously that the leucine to proline residue substitution at position 189 (L189P) in barley HvHKT1;5 disrupts its characteristic plasma membrane localisation and Na+ conductance. Here, we focus on a surprising observation that a single residue deletion of methionine at position 372 (M372del) within the conserved VMMYL motif in plant HKTs, restores plasma membrane localisation but not Na+ conductance in HvHKT1;5 P189. To clarify why the singular M372 deletion regains plasma membrane localisation, we built 3D models and defined α-helical assembly pathways of the P189 M372del mutant, and compared these findings to the wild-type protein, and the HvHKT1;5 L189 variant and its M372del mutant. We find that α-helical association and assembly pathways in HvHKT1;5 proteins fall in two contrasting categories. Inspections of structural flexibility through molecular dynamics simulations revealed that the conformational states of HvHKT1;5 P189 diverge from those of the L189 variant and M372del mutants. We propose that M372del in HvHKT1;5 P189 instigates structural rearrangements allowing routing to the plasma membrane, while the restoration of conductance would require further interventions. We integrate the microscopy, electrophysiology, and biocomputational data and discuss how a profound structural change in HvHKT1;5 P189 M372del impacts its α-helical protein association pathway and flexibility, and how these features underlie a delicate balance leading to restoring plasma membrane localisation but not Na+ conductance.


Asunto(s)
Eliminación de Gen , Genes de Plantas , Hordeum/genética , Mutación , Proteínas de Plantas/genética , Sodio/metabolismo , Secuencias de Aminoácidos , Membrana Celular/metabolismo , Hordeum/metabolismo , Proteínas de Plantas/química
18.
Biochem Soc Trans ; 38(2): 683-8, 2010 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-20298243

RESUMEN

The identification of genes underlying complex quantitative traits such as grain yield by means of conventional genetic analysis (positional cloning) requires the development of several large mapping populations. However, it is possible that phenotypically related, but more extreme, allelic variants generated by mutational studies could provide a means for more efficient cloning of QTLs (quantitative trait loci). In barley (Hordeum vulgare), with the development of high-throughput genome analysis tools, efficient genome-wide identification of genetic loci harbouring mutant alleles has recently become possible. Genotypic data from NILs (near-isogenic lines) that carry induced or natural variants of genes that control aspects of plant development can be compared with the location of QTLs to potentially identify candidate genes for development--related traits such as grain yield. As yield itself can be divided into a number of allometric component traits such as tillers per plant, kernels per spike and kernel size, mutant alleles that both affect these traits and are located within the confidence intervals for major yield QTLs may represent extreme variants of the underlying genes. In addition, the development of detailed comparative genomic models based on the alignment of a high-density barley gene map with the rice and sorghum physical maps, has enabled an informed prioritization of 'known function' genes as candidates for both QTLs and induced mutant genes.


Asunto(s)
Clonación Molecular/métodos , Hordeum/genética , Mutagénesis/fisiología , Plantas Modificadas Genéticamente/genética , Sitios de Carácter Cuantitativo/genética , Modelos Biológicos , Modelos Genéticos , Carácter Cuantitativo Heredable
19.
Commun Biol ; 3(1): 258, 2020 05 22.
Artículo en Inglés | MEDLINE | ID: mdl-32444849

RESUMEN

During plant growth, sodium (Na+) in the soil is transported via the xylem from the root to the shoot. While excess Na+ is toxic to most plants, non-toxic concentrations have been shown to improve crop yields under certain conditions, such as when soil K+ is low. We quantified grain Na+ across a barley genome-wide association study panel grown under non-saline conditions and identified variants of a Class 1 HIGH-AFFINITY-POTASSIUM-TRANSPORTER (HvHKT1;5)-encoding gene responsible for Na+ content variation under these conditions. A leucine to proline substitution at position 189 (L189P) in HvHKT1;5 disturbs its characteristic plasma membrane localisation and disrupts Na+ transport. Under low and moderate soil Na+, genotypes containing HvHKT1:5P189 accumulate high concentrations of Na+ but exhibit no evidence of toxicity. As the frequency of HvHKT1:5P189 increases significantly in cultivated European germplasm, we cautiously speculate that this non-functional variant may enhance yield potential in non-saline environments, possibly by offsetting limitations of low available K+.


Asunto(s)
Proteínas de Transporte de Catión/metabolismo , Regulación de la Expresión Génica de las Plantas , Hordeum/metabolismo , Proteínas de Plantas/metabolismo , Raíces de Plantas/metabolismo , Brotes de la Planta/metabolismo , Sodio/metabolismo , Proteínas de Transporte de Catión/genética , Estudio de Asociación del Genoma Completo , Hordeum/genética , Hordeum/crecimiento & desarrollo , Proteínas de Plantas/genética , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Brotes de la Planta/genética , Brotes de la Planta/crecimiento & desarrollo
20.
Sci Rep ; 9(1): 17250, 2019 11 21.
Artículo en Inglés | MEDLINE | ID: mdl-31754200

RESUMEN

The composition of plant cell walls is important in determining cereal end uses. Unlike other widely consumed cereal grains barley is comparatively rich in (1,3;1,4)-ß-glucan, a source of dietary fibre. Previous work showed Cellulose synthase-like genes synthesise (1,3;1,4)-ß-glucan in several tissues. HvCslF6 encodes a grain (1,3;1,4)-ß-glucan synthase, whereas the function of HvCslF9 is unknown. Here, the relationship between mRNA levels of HvCslF6, HvCslF9, HvGlbI (1,3;1,4)-ß-glucan endohydrolase, and (1,3;1,4)-ß-glucan content was studied in developing grains of four barley cultivars. HvCslF6 was differentially expressed during mid (8-15 DPA) and late (38 DPA) grain development stages while HvCslF9 transcript was only clearly detected at 8-10 DPA. A peak of HvGlbI expression was detected at 15 DPA. Differences in transcript abundance across the three genes could partially explain variation in grain (1,3;1,4)-ß-glucan content in these genotypes. Remarkably narrow sequence variation was found within the HvCslF6 promoter and coding sequence and does not explain variation in (1,3;1,4)-ß-glucan content. Our data emphasise the genotype-dependent accumulation of (1,3;1,4)-ß-glucan during barley grain development and a role for the balance between hydrolysis and synthesis in determining (1,3;1,4)-ß-glucan content, and suggests that other regulatory sequences or proteins are likely to be involved in this trait in developing grain.


Asunto(s)
Variación Genética/genética , Glucosiltransferasas/genética , Hordeum/genética , Hordeum/metabolismo , Proteínas de Plantas/genética , beta-Glucanos/metabolismo , Pared Celular/genética , Pared Celular/metabolismo , Fibras de la Dieta/metabolismo , Grano Comestible/genética , Grano Comestible/metabolismo , Genotipo , Glucosiltransferasas/metabolismo , Fenotipo , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Regiones Promotoras Genéticas/genética , Semillas/genética , Semillas/metabolismo
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