title: Loss of Cellulose synthase-like F6 function affects mixed-linkage glucan deposition, cell wall mechanical properties, and defense responses in vegetative tissues of rice.
abstract: Mixed-linkage glucan (MLG) is a cell wall polysaccharide containing a backbone of unbranched (1,3)- and (1,4)-linked β-glucosyl residues. Based on its occurrence in plants and chemical characteristics, MLG has primarily been associated with the regulation of cell wall expansion due to its high and transient accumulation in young, expanding tissues. The Cellulose synthase-like F (CslF) subfamily of glycosyltransferases has previously been implicated in mediating the biosynthesis of this polymer. We confirmed that the rice (Oryza sativa) CslF6 gene mediates the biosynthesis of MLG by overexpressing it in Nicotiana benthamiana. Rice cslf6 knockout mutants show a slight decrease in height and stem diameter but otherwise grew normally during vegetative development. However, cslf6 mutants display a drastic decrease in MLG content (97% reduction in coleoptiles and virtually undetectable in other tissues). Immunodetection with an anti-MLG monoclonal antibody revealed that the coleoptiles and leaves retain trace amounts of MLG only in specific cell types such as sclerenchyma fibers. These results correlate with the absence of endogenous MLG synthase activity in mutant seedlings and 4-week-old sheaths. Mutant cell walls are weaker in mature stems but not seedlings, and more brittle in both stems and seedlings, compared to wild type. Mutants also display lesion mimic phenotypes in leaves, which correlates with enhanced defense-related gene expression and enhanced disease resistance. Taken together, our results underline a weaker role of MLG in cell expansion than previously thought, and highlight a structural role for MLG in nonexpanding, mature stem tissues in rice.
authors.affiliation: Joint BioEnergy Institute, Emeryville, California 94608, USA.
authors.forename: Miguel E
authors.initials: ME
authors.lastname: Vega-Sánchez
authors.forename: Yves
authors.initials: Y
authors.lastname: Verhertbruggen
authors.forename: Ulla
authors.initials: U
authors.lastname: Christensen
authors.forename: Xuewei
authors.initials: X
authors.lastname: Chen
authors.forename: Vaishali
authors.initials: V
authors.lastname: Sharma
authors.forename: Patanjali
authors.initials: P
authors.lastname: Varanasi
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Jobling
authors.forename: Mark
authors.initials: M
authors.lastname: Talbot
authors.forename: Rosemary G
authors.initials: RG
authors.lastname: White
authors.forename: Michael
authors.initials: M
authors.lastname: Joo
authors.forename: Seema
authors.initials: S
authors.lastname: Singh
authors.forename: Manfred
authors.initials: M
authors.lastname: Auer
authors.forename: Henrik V
authors.initials: HV
authors.lastname: Scheller
authors.forename: Pamela C
authors.initials: PC
authors.lastname: Ronald
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism*; D004789:Enzyme Activation; D018506:Gene Expression Regulation, Plant; D055786:Gene Knockout Techniques; D017343:Genes, Plant; D005936:Glucans / Q000378:metabolism*; D005964:Glucosyltransferases / Q000378:metabolism*; D009154:Mutation; D012275:Oryza / Q000201:enzymology* / Q000235:genetics / Q000382:microbiology; D010935:Plant Diseases / Q000276:immunology / Q000382:microbiology; D057865:Plant Immunity; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018547:Plant Stems / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism / Q000382:microbiology; D036226:Seedlings / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014974:Xanthomonas / Q000276:immunology / Q000472:pathogenicity
chemical_list: D000911:Antibodies, Monoclonal; D005936:Glucans; D010940:Plant Proteins; D005964:Glucosyltransferases; C478648:cellulose synthase