The protein structure of the cellulose synthase-like protein(CSL) was similar to cellulose synthase(CesA),including the conservative sequence D,D,D,QXXRW.One full-length cDNA of the cellulose synthase-like protein D(C...The protein structure of the cellulose synthase-like protein(CSL) was similar to cellulose synthase(CesA),including the conservative sequence D,D,D,QXXRW.One full-length cDNA of the cellulose synthase-like protein D(CslD) gene was cloned by reverse transcriptase(RT)-polymerase chain reaction(PCR) with 5’,3’ rapid amplification of cDNA ends(RACE) methods using degenerate primers designed from the homologous sequences of the CesA genes.A multiple comparison sequence analysis was conducted concurrently with bioinformatic methods to analyze the obtained sequence.Results of the sequence analysis showed that this cDNA was 4 150 bp in length and contained a single open reading frame encoding a protein of 1 132 amino acids.The multiple comparison sequence analysis showed that the deduced amino acid sequence shared high similarity (over 71%) with the ClCslD genes from Populus tremuloides,Oryza sativa,and Arabidopsis thaliana. This work will help lay an important foundation for further molecular studies with cellulose synthesis of plants.展开更多
The plant cell wall serves as a barrier in defense against pathogen invasion.However,the specific contribution of cell walls in vascular tissues to plant immunity remains largely unexplored.In this study,we demonstrat...The plant cell wall serves as a barrier in defense against pathogen invasion.However,the specific contribution of cell walls in vascular tissues to plant immunity remains largely unexplored.In this study,we demonstrate that OsCSLC3,a member of the rice cellulose synthase-like(CSL)gene family,is predominantly expressed in vascular tissues and that its overexpression promotes hemicellulose biosynthesis.This enhancement of hemicellulose accumulation is associated with improved disease resistance.Targeted editing of conserved cis-regulatory elements in the OsCSLC35′untranslated region(UTR)showed that deletion of the specific fragment(−575 to−824 bp)elevated OsCSLC3 transcript levels,promoted hemicellulose accumulation,enhanced disease resistance,and improved agronomic traits.Our findings highlight a previously underappreciated role for hemicellulose in plant immunity and demonstrate that precise 5′UTR editing is a promising strategy for improving disease resistance and agronomic traits.展开更多
In cereals, the presence of soluble polysaccharides including（1,3;1,4）-b-glucan has downstream implications for human health, animal feed and biofuel applications. Sorghum bicolor（L.） Moench is a versatile crop, b...In cereals, the presence of soluble polysaccharides including（1,3;1,4）-b-glucan has downstream implications for human health, animal feed and biofuel applications. Sorghum bicolor（L.） Moench is a versatile crop, but there are limited reports regarding the content of such soluble polysaccharides.Here, the amount of（1,3;1,4）-b-glucan present in sorghum tissues was measured using a Megazyme assay. Very low amounts were present in the grain, ranging from 0.16%–0.27%（w/w）, while there was a greater quantity in vegetative tissues at 0.12–1.71%（w/w）. The fine structure of（1,3;1,4）-b-glucan, as denoted by the ratio of cellotriosyl and cellotetraosyl residues,was assessed by high performance liquid chromatography（HPLC） and ranged from 2.6–3：1 in the grain, while ratios in vegetative tissues were lower at 2.1–2.6：1. The distribution of（1,3;1,4）-b-glucan was examined using a specific antibody and observed with fl uorescence and transmission electron microscopy. Micrographs showed a variable distribution of（1,3;1,4）-b-glucan in fl uenced by temporal and spatial factors. The sorghum orthologs of genes implicated in the synthesis of（1,3;1,4）-b-glucan in other cereals, such as the Cellulose synthase-like（Csl） F and H gene families were de fined.Transcript pro filing of these genes across sorghum tissues was carried out using real-time quantitative polymerase chain reaction, indicating that, as in other cereals, Csl F6 transcripts dominated.展开更多
The CELLULOSE SYNTHASE-LIKE C （CSLC） family is an ancient lineage within the CELLULOSE SYNTHASE/CELLULOSE SYNTHASE-LIKE （CESA/CSL） polysaccharide synthase superfamily that is thought to have arisen before the dive...The CELLULOSE SYNTHASE-LIKE C （CSLC） family is an ancient lineage within the CELLULOSE SYNTHASE/CELLULOSE SYNTHASE-LIKE （CESA/CSL） polysaccharide synthase superfamily that is thought to have arisen before the divergence of mosses and vascular plants. As studies in the flowering plant Arabidopsis have suggested synthesis of the （1,4）-β-glucan backbone of xyloglucan （XyG）, a wall polysaccharide that tethers adjacent cellulose microfibrils to each other, as a probable function for the CSLCs, CSLC function was investigated in barley （Hordeum vulgate L.）, a species with low amounts of XyG in its walls. Four barley CSLCgenes were identified （designated HvCSLC1-4）. Phylogenetic analysis reveals three well supported clades of CSLCs in flowering plants, with barley having representatives in two of these clades. The four barley CSLCs were expressed in various tissues, with in situ PCR detecting transcripts in all cell types of the coleoptile and root, including cells with primary and secondary cell walls. Co-expression analysis showed that HvCSLC3 was coordinately expressed with putative XyG xylosyltransferase genes. Both immuno-EM and membrane fractionation showed that HvCSLC2 was located in the plasma membrane of barley suspension-cultured cells and was not in internal membranes such as endoplasmic reticulum or Golgi apparatus. Based on our current knowledge of the sub-cellular locations of polysaccharide synthesis, we conclude that the CSLC family probably contains more than one type of polysaccharide synthase.展开更多
文摘The protein structure of the cellulose synthase-like protein(CSL) was similar to cellulose synthase(CesA),including the conservative sequence D,D,D,QXXRW.One full-length cDNA of the cellulose synthase-like protein D(CslD) gene was cloned by reverse transcriptase(RT)-polymerase chain reaction(PCR) with 5’,3’ rapid amplification of cDNA ends(RACE) methods using degenerate primers designed from the homologous sequences of the CesA genes.A multiple comparison sequence analysis was conducted concurrently with bioinformatic methods to analyze the obtained sequence.Results of the sequence analysis showed that this cDNA was 4 150 bp in length and contained a single open reading frame encoding a protein of 1 132 amino acids.The multiple comparison sequence analysis showed that the deduced amino acid sequence shared high similarity (over 71%) with the ClCslD genes from Populus tremuloides,Oryza sativa,and Arabidopsis thaliana. This work will help lay an important foundation for further molecular studies with cellulose synthesis of plants.
基金supported by the Major Projects in Agricultural Biological Breeding(2022ZD04002)to Xuewei ChenNational Natural Science Foundation of China(32121003 and 32425005)to Xuewei Chen+3 种基金(32172419 and 32372555)to Weitao LiCornerstone Science Foundation through the XPLORER PRIZE and New Cornerstone Investigator Program to Xuewei ChenSichuan Science and Technology Program(2023NSFSC0005)to Xuewei Chen,(2023NSFSC1996)to Weitao Li,(2023NSFSC0155)to Qingqing Hou(2024YFNH0014)to Junjie Yin.
文摘The plant cell wall serves as a barrier in defense against pathogen invasion.However,the specific contribution of cell walls in vascular tissues to plant immunity remains largely unexplored.In this study,we demonstrate that OsCSLC3,a member of the rice cellulose synthase-like(CSL)gene family,is predominantly expressed in vascular tissues and that its overexpression promotes hemicellulose biosynthesis.This enhancement of hemicellulose accumulation is associated with improved disease resistance.Targeted editing of conserved cis-regulatory elements in the OsCSLC35′untranslated region(UTR)showed that deletion of the specific fragment(−575 to−824 bp)elevated OsCSLC3 transcript levels,promoted hemicellulose accumulation,enhanced disease resistance,and improved agronomic traits.Our findings highlight a previously underappreciated role for hemicellulose in plant immunity and demonstrate that precise 5′UTR editing is a promising strategy for improving disease resistance and agronomic traits.
基金supported by funding from the Australian Development Scholarship (ADS),Australia Awards and the Australian Research Council Centre of Excellence in Plant Cell Walls,The University of Adelaide
文摘In cereals, the presence of soluble polysaccharides including（1,3;1,4）-b-glucan has downstream implications for human health, animal feed and biofuel applications. Sorghum bicolor（L.） Moench is a versatile crop, but there are limited reports regarding the content of such soluble polysaccharides.Here, the amount of（1,3;1,4）-b-glucan present in sorghum tissues was measured using a Megazyme assay. Very low amounts were present in the grain, ranging from 0.16%–0.27%（w/w）, while there was a greater quantity in vegetative tissues at 0.12–1.71%（w/w）. The fine structure of（1,3;1,4）-b-glucan, as denoted by the ratio of cellotriosyl and cellotetraosyl residues,was assessed by high performance liquid chromatography（HPLC） and ranged from 2.6–3：1 in the grain, while ratios in vegetative tissues were lower at 2.1–2.6：1. The distribution of（1,3;1,4）-b-glucan was examined using a specific antibody and observed with fl uorescence and transmission electron microscopy. Micrographs showed a variable distribution of（1,3;1,4）-b-glucan in fl uenced by temporal and spatial factors. The sorghum orthologs of genes implicated in the synthesis of（1,3;1,4）-b-glucan in other cereals, such as the Cellulose synthase-like（Csl） F and H gene families were de fined.Transcript pro filing of these genes across sorghum tissues was carried out using real-time quantitative polymerase chain reaction, indicating that, as in other cereals, Csl F6 transcripts dominated.
文摘The CELLULOSE SYNTHASE-LIKE C （CSLC） family is an ancient lineage within the CELLULOSE SYNTHASE/CELLULOSE SYNTHASE-LIKE （CESA/CSL） polysaccharide synthase superfamily that is thought to have arisen before the divergence of mosses and vascular plants. As studies in the flowering plant Arabidopsis have suggested synthesis of the （1,4）-β-glucan backbone of xyloglucan （XyG）, a wall polysaccharide that tethers adjacent cellulose microfibrils to each other, as a probable function for the CSLCs, CSLC function was investigated in barley （Hordeum vulgate L.）, a species with low amounts of XyG in its walls. Four barley CSLCgenes were identified （designated HvCSLC1-4）. Phylogenetic analysis reveals three well supported clades of CSLCs in flowering plants, with barley having representatives in two of these clades. The four barley CSLCs were expressed in various tissues, with in situ PCR detecting transcripts in all cell types of the coleoptile and root, including cells with primary and secondary cell walls. Co-expression analysis showed that HvCSLC3 was coordinately expressed with putative XyG xylosyltransferase genes. Both immuno-EM and membrane fractionation showed that HvCSLC2 was located in the plasma membrane of barley suspension-cultured cells and was not in internal membranes such as endoplasmic reticulum or Golgi apparatus. Based on our current knowledge of the sub-cellular locations of polysaccharide synthesis, we conclude that the CSLC family probably contains more than one type of polysaccharide synthase.
