A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-β-d-glucan synthesis in transgenic Arabidopsis Export

@article{citeulike:4364906, abstract = {10.1073/pnas.0902019106 The walls of grasses and related members of the Poales are characterized by the presence of the polysaccharide (,, ,)-β-D-glucan (β-glucan). To date, only members of the grass-specific () gene family have been implicated in its synthesis. Assuming that other grass-specific genes also might encode synthases for this polysaccharide, we cloned , a gene from barley ( L.), and expressed an epitope-tagged version of the cDNA in , a species with no genes and no β-glucan in its walls. Transgenic lines that had detectable amounts of the epitope-tagged HvCSLH1 protein accumulated β-glucan in their walls. The presence of β-glucan was confirmed by immunoelectron microscopy (immuno-EM) of sectioned tissues and chemical analysis of wall extracts. In the chemical analysis, characteristic tri- and tetra-saccharides were identified by high-performance anion-exchange chromatography and MALDI-TOF MS following their release from transgenic walls by a specific β-glucan hydrolase. Immuno-EM also was used to show that the epitope-tagged HvCSLH1 protein was in the endoplasmic reticulum and Golgi-associated vesicles, but not in the plasma membrane. In barley, was expressed at very low levels in leaf, floral tissues, and the developing grain. In leaf, expression was highest in xylem and interfascicular fiber cells that have walls with secondary thickenings containing β-glucan. Thus both the and families contribute to β-glucan synthesis in grasses and probably do so independently of each other, because there is no significant transcriptional correlation between these genes in the barley tissues surveyed.}, author = {Doblin, Monika S. and Pettolino, Filomena A. and Wilson, Sarah M. and Campbell, Rebecca and Burton, Rachel A. and Fincher, Geoffrey B. and Newbigin, Ed and Bacic, Antony}, citeulike-article-id = {4364906}, citeulike-linkout-0 = {http://dx.doi.org/10.1073/pnas.0902019106}, citeulike-linkout-1 = {http://www.pnas.org/content/0902019106v1//.abstract}, citeulike-linkout-2 = {http://www.pnas.org/content/0902019106v1//.full.pdf}, citeulike-linkout-3 = {http://view.ncbi.nlm.nih.gov/pubmed/19321749}, citeulike-linkout-4 = {http://www.hubmed.org/display.cgi?uids=19321749}, day = {7}, doi = {10.1073/pnas.0902019106}, journal = {Proceedings of the National Academy of Sciences}, keywords = {cesa}, month = {April}, number = {14}, pages = {5996--6001}, posted-at = {2009-04-19 15:01:04}, priority = {2}, title = {A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-β-d-glucan synthesis in transgenic Arabidopsis}, url = {http://dx.doi.org/10.1073/pnas.0902019106}, volume = {106}, year = {2009} }

TY - JOUR ID - citeulike:4364906 L3 - citeulike-article-id:4364906 N2 - 10.1073/pnas.0902019106 The walls of grasses and related members of the Poales are characterized by the presence of the polysaccharide (,, ,)-β-D-glucan (β-glucan). To date, only members of the grass-specific () gene family have been implicated in its synthesis. Assuming that other grass-specific genes also might encode synthases for this polysaccharide, we cloned , a gene from barley ( L.), and expressed an epitope-tagged version of the cDNA in , a species with no genes and no β-glucan in its walls. Transgenic lines that had detectable amounts of the epitope-tagged HvCSLH1 protein accumulated β-glucan in their walls. The presence of β-glucan was confirmed by immunoelectron microscopy (immuno-EM) of sectioned tissues and chemical analysis of wall extracts. In the chemical analysis, characteristic tri- and tetra-saccharides were identified by high-performance anion-exchange chromatography and MALDI-TOF MS following their release from transgenic walls by a specific β-glucan hydrolase. Immuno-EM also was used to show that the epitope-tagged HvCSLH1 protein was in the endoplasmic reticulum and Golgi-associated vesicles, but not in the plasma membrane. In barley, was expressed at very low levels in leaf, floral tissues, and the developing grain. In leaf, expression was highest in xylem and interfascicular fiber cells that have walls with secondary thickenings containing β-glucan. Thus both the and families contribute to β-glucan synthesis in grasses and probably do so independently of each other, because there is no significant transcriptional correlation between these genes in the barley tissues surveyed. IS - 14 JF - Proceedings of the National Academy of Sciences EP - 6001 TI - A barley cellulose synthase-like CSLH gene mediates (1,3;1,4)-β-d-glucan synthesis in transgenic Arabidopsis VL - 106 SP - 5996 KW - cesa AU - Doblin, Monika AU - Pettolino, Filomena AU - Wilson, Sarah AU - Campbell, Rebecca AU - Burton, Rachel AU - Fincher, Geoffrey AU - Newbigin, Ed AU - Bacic, Antony PY - 2009/04/07/ UR - http://dx.doi.org/10.1073/pnas.0902019106 ER -