Disruption of ATCSLD5 results in reduced growth, reduced xylan and homogalacturonan synthase activity and altered xylan occurrence in Arabidopsis
Disruption of ATCSLD5 results in reduced growth, reduced xylan and homogalacturonan synthase activity and altered xylan occurrence in Arabidopsis
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DOI：
10.1111/j.1365-313x.2007.03281.x
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发表时间：
2007-12-01
期刊：
影响因子：
7.2
通讯作者：
Willats, William G. T.
中科院分区：
文献类型：
--
作者：
Bernal, Adriana Jimena;Jensen, Jakob Kruger;Willats, William G. T.
纤维素酶样基因（CSLs）是一类编码糖基转移酶（GTs）的基因，参与植物细胞壁的生物合成。已知CSLA和CSLF家族分别含有甘露聚糖和葡聚糖糖苷酶，但其他CSL的产物未知。在这里，我们报告了在拟南芥中破坏ATCSLD 5表达的影响。在ATCSLD 5基因敲除的植物中，茎和根的生长都显著降低，并且这些植物对纤维素合成酶抑制剂异恶酰草胺的敏感性也增加。抗体和碳水化合物结合模块标记表明，木聚糖在茎的水平降低，并在体外GT测定使用微粒体从茎显示，ATCSLD 5敲除植物也有木聚糖和homogalacturonan合成酶活性降低。ATCSLD 5和ATCSLD 3在本氏烟草（Nicotiana benthamiana）中的表达，在C-或N-末端荧光标记，表明这些GT可能定位于高尔基体中。然而，荧光标记的位置影响了这两种蛋白质的亚细胞定位。所提出的工作提供了在植物中破坏ATCSLD 5的影响的全面分析，并讨论了该基因和其他ATCSLDs在细胞壁生物合成中的可能作用。
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.