Samples of ten winter wheat varieties, grown at four locations on the eastern side of the UK were assessed for alcohol yield. There was a strong negative association between alcohol yield and grain nitrogen content, but this was strongly influenced by differences between the sites, with a range of alcohol yields observed at given nitrogen levels. Inclusion of grain size parameters, thousand grain weight and grain length:width ratio, along with nitrogen, in a multiple regression, enabled more accurate prediction of alcohol yield for most of the varieties, indicating that grain size would be a useful parameter for selecting the best samples within varieties. A small-scale measurement of hot water extract, along with grain nitrogen content, gave a reasonable prediction of alcohol yield, but the regression equation was not consistent across seasons. Starch content and turbidity (a measure of the ease of starch release) were both shown to be significantly influenced by variety, but not to give significant correlations with alcohol yield over a range of samples. Positive expression of these characters appeared, however, to contribute to improved alcohol yield in certain of the varieties. It was concluded that different genetic mechanisms influence alcohol yield and that improved breeding lines could be developed by combining positive alcohol yield attributes from complementary parents.
Barley (Hordeum vulgare L) grain is comparatively rich in (1,3;1,4)-b-glucan, a source of fermentable dietary fibre that protects against various human health conditions. However, low grain (1,3;1,4)-b-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)-b-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)-b-glucan content. (1,3;1,4)-b-Glucan was absent in the grain of cslf6 knockout lines, whereas cslf9 knockout lines had similar (1,3;1,4)-b-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)-b-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)-b-glucan in mature grain.
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+.
We quantified grain sodium (Na + ) content across a barley GWAS panel grown under optimal conditions. We identified a strong association with a region containing two low and one high Na + accumulating haplotypes of a Class 1 HIGH-AFFINITY POTASSIUM TRANSPORTER (HKT1;5) known to be involved in regulating plant Na + homeostasis. The haplotypes exhibited an average 1.8-fold difference in grain Na + content. We show that an L189P substitution disrupts Na + transport in the high Na + lines, disturbs the plasma membrane localisation typical of HKT1;5 and induces a conformational change in the protein predicted to compromise function. Under NaCl stress, lines containing P189 accumulate high levels of Na + , but show no significant difference in biomass. P189 increases in frequency from wildspecies to elite cultivars leading us to speculate that the compromised haplotype is undergoing directional selection possibly due to the value of Na + as a functional nutrient in non-saline environments.
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