Disruption of ATCSLD5 results in reduced growth, reduced xylan and homogalacturonan synthase activity and altered xylan occurrence in Arabidopsis.
Members of a large family of cellulose synthase-like genes (CSLs) are predicted to encode glycosyl transferases (GTs) involved in the biosynthesis of plant cell walls. The CSLA and CSLF families are known to contain mannan and glucan synthases, respectively, but the products of other CSLs are unknown. Here we report the effects of disrupting ATCSLD5 expression in Arabidopsis. Both stem and root growth were significantly reduced in ATCSLD5 knock-out plants, and these plants also had increased susceptibility to the cellulose synthase inhibitor isoxaben. Antibody and carbohydrate-binding module labelling indicated a reduction in the level of xylan in stems, and in vitro GT assays using microsomes from stems revealed that ATCSLD5 knock-out plants also had reduced xylan and homogalacturonan synthase activity. Expression in Nicotiana benthamiana of ATCSLD5 and ATCSLD3, fluorescently tagged at either the C- or the N-terminal, indicated that these GTs are likely to be localized in the Golgi apparatus. However, the position of the fluorescent tag affected the subcellular localization of both proteins. The work presented provides a comprehensive analysis of the effects of disrupting ATCSLD5 in planta, and the possible role(s) of this gene and other ATCSLDs in cell wall biosynthesis are discussed.
Bernal
Adriana Jimena
AJ
Department of Molecular Biology, Københavns Biocenter, Københavns Universitet, Ole Maaløes Vej 5, 2200 København N, Denmark.
Jensen
Jakob Krüger
JK
Harholt
Jesper
J
Sørensen
Susanne
S
Moller
Isabel
I
Blaukopf
Claudia
C
Johansen
Bo
B
de Lotto
Robert
R
Pauly
Markus
M
Scheller
Henrik Vibe
HV
Willats
William G T
WG
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D029681:Arabidopsis Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D001549:Benzamides / Q000494:pharmacology; D005964:Glucosyltransferases / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D005966:Glucuronidase / Q000032:analysis; D010368:Pectins / Q000096:biosynthesis; D010430:Pentosyltransferases / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics; D014990:Xylans / Q000378:metabolism*
D029681:Arabidopsis Proteins; D001549:Benzamides; D014990:Xylans; C057467:isoxaben; D010368:Pectins; C526694:CSLD5 protein, Arabidopsis; D005964:Glucosyltransferases; C478648:cellulose synthase; D010430:Pentosyltransferases; C038331:1,4-beta-D-xylan synthase; D005966:Glucuronidase; C003181:polygalacturonic acid