Overexpression of HvCslF6 in barley grain alters carbohydrate partitioning plus transfer tissue and endosperm development

Lim, Wai Li; Collins, Helen; Byrt, Caitlin S.; Lahnstein, Jelle; Shirley, Neil J.; Aubert, Matthew K.; Tucker, Matthew R.; Peukert, Manuela; Matros, Andrea; Burton, Rachel A.

doi:10.1093/jxb/erz407

Cited by 20 publications

(9 citation statements)

References 78 publications

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“…The novelty in the present studies is the influence of β-glucan. The negative correlation between starch content and β-glucan content is well documented and explained (Lim et al, 2020). It is found here that pasting viscosities significantly and positively correlate to β-glucan content (Table 2).…”

Section: Discussionsupporting

confidence: 79%

The contribution of β-glucan and starch fine structure to texture of oat-fortified wheat noodles

et al. 2020

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Wheat flour noodles are sometimes fortified with β-glucan for nutritional value, but this can decrease eating quality. The contributions of β-glucan and starch molecular fine structure to physicochemical properties of wholemeal oat flour and to the texture of oat-fortified white salted noodles were investigated here. Hardness of oat-fortified noodles was controlled by the longer amylopectin chains (DP ≥ 26) and amount of longer amylose chains (DP ≥ 1000). Higher levels of β-glucan, in the range from 3.1 to 5.2%, result in increased noodle hardness. Pasting viscosities of wholemeal oat flour positively correlate with the hardness of oat-fortified noodles. The swelling power of oat flour is not correlated with either pasting viscosities of oat flour or noodle hardness. Longer amylopectin chains and the amount of longer amylose chains both control the pasting viscosities of oat flour, which in turn affect noodle texture. This provides new means, based on starch and β-glucan molecular structure, to choose oats with optimal starch structure and β-glucan content for targeted oat-fortified noodle quality.

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Section: Discussionsupporting

confidence: 79%

The contribution of β-glucan and starch fine structure to texture of oat-fortified wheat noodles

et al. 2020

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“…Dysfunction of these cells seems to cause impaired assimilate flow and/or signal transmission from maternal tissues, resulting in patterning defects specifically in adjacent endosperm regions. Similar effects on barley endosperm differentiation probably caused by non-functional ETCs and impaired transport were observed by overexpression of HvCeSLF6 (Wai et al, 2019). Subsequently, ETCs serve as a communication hub that integrates differentiation processes in filial grain organs.…”

Section: Development Of the Transfer Cell Anatomy Is A Prerequisite Fmentioning

confidence: 54%

Barley HISTIDINE KINASE 1 (HvHK1) coordinates transfer cell specification in the young endosperm

et al. 2020

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SUMMARY Cereal endosperm represents the most important source of the world’s food; nevertheless, the molecular mechanisms underlying cell and tissue differentiation in cereal grains remain poorly understood. Endosperm cellularization commences at the maternal–filial intersection of grains and generates endosperm transfer cells (ETCs), a cell type with a prominent anatomy optimized for efficient nutrient transport. Barley HISTIDINE KINASE1 (HvHK1) was identified as a receptor component with spatially restricted expression in the syncytial endosperm where ETCs emerge. Here, we demonstrate its function in ETC fate acquisition using RNA interference‐mediated downregulation of HvHK1. Repression of HvHK1 impairs cell specification in the central ETC region and the development of transfer cell morphology, and consecutively defects differentiation of adjacent endosperm tissues. Coinciding with reduced expression of HvHK1, disturbed cell plate formation and fusion were observed at the initiation of endosperm cellularization, revealing that HvHK1 triggers initial cytokinesis of ETCs. Cell‐type‐specific RNA sequencing confirmed loss of transfer cell identity, compromised cell wall biogenesis and reduced transport capacities in aberrant cells and elucidated two‐component signaling and hormone pathways that are mediated by HvHK1. Gene regulatory network modeling was used to specify the direct targets of HvHK1; this predicted non‐canonical auxin signaling elements as the main regulatory links governing cellularization of ETCs, potentially through interaction with type‐B response regulators. This work provides clues to previously unknown molecular mechanisms directing ETC specification, a process with fundamental impact on grain yield in cereals.

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“…Immunohistochemical detection of (1,3;1,4)‐β‐glucan confirmed the absence of this polysaccharide in mature grain of cslf6‐2 homozygous mutants, whereas a weak labelling was detected across aleurone and endosperm tissues in heterozygous lines. It is known that an inverse relationship exists between (1,3;1,4)‐β‐glucan and starch content in grains of several grass species, including barley, based on comparative studies across cereal species with contrasting levels of this polysaccharide (Trafford et al ., 2013; Lim et al ., 2019). The precise mechanism/s that regulate this relationship remain unknown.…”

Section: Discussionmentioning

confidence: 99%

Targeted mutation of barley (1,3;1,4)‐β‐glucan synthases reveals complex relationships between the storage and cell wall polysaccharide content

et al. 2020

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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.

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Overexpression of HvCslF6 in barley grain alters carbohydrate partitioning plus transfer tissue and endosperm development

Abstract: Overexpression of the HvCslF6 gene in hull-less barley grain perturbs sucrose uptake and allocation, and impairs transfer tissue and endosperm development

Cited by 20 publications

References 78 publications

The contribution of β-glucan and starch fine structure to texture of oat-fortified wheat noodles

The contribution of β-glucan and starch fine structure to texture of oat-fortified wheat noodles

Barley HISTIDINE KINASE 1 (HvHK1) coordinates transfer cell specification in the young endosperm

Targeted mutation of barley (1,3;1,4)‐β‐glucan synthases reveals complex relationships between the storage and cell wall polysaccharide content

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