UGT74D1 is a novel auxin glycosyltransferase from Arabidopsis thaliana.
UGT74D1 is a novel auxin glycosyltransferase from Arabidopsis thaliana.
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DOI：
10.1371/journal.pone.0061705
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Hou BK
中科院分区：
文献类型：
--
作者：
Jin SH;Ma XM;Han P;Wang B;Sun YG;Zhang GZ;Li YJ;Hou BK
生长素是一类在植物生长发育过程中起重要作用的激素。生长素的糖基化被认为是控制活性生长素水平的重要机制。因此，鉴定生长素糖基转移酶对进一步了解生长素的调控具有重要意义。在这项研究中，我们生化筛选拟南芥糖基转移酶超家族的L组对生长素的酶活性。UGT 74 D1是一种新的生长素糖基转移酶。通过对生长素等8种底物的体外反应产物进行HPLC和LC-MS分析，发现UGT 74 D1对吲哚-3-丁酸[IBA]、吲哚-3-丙酸[IPA]、吲哚-3-乙酸[IAA]和萘乙酸[NAA]具有较强的葡萄糖基化活性，催化它们形成相应的葡萄糖酯。生化特性表明，该酶在pH 6.0的HEPES缓冲液和37°C下具有最大活性。此外，还对转UGT 74 D1基因拟南芥的粗蛋白进行了酶活性分析，并对转UGT 74 D1基因拟南芥的IBA代谢产物进行了分析。实验结果表明，在植物中过量生产UGT 74 D1确实导致IBA的葡萄糖缀合物水平增加。此外，UGT 74 D1过表达株系显示卷曲叶表型，表明UGT 74 D1在影响生长素活性方面的生理作用。本研究为进一步研究植物生长素糖基化调控提供了新的靶基因。
Auxin is one type of phytohormones that plays important roles in nearly all aspects of plant growth and developmental processes. The glycosylation of auxins is considered to be an essential mechanism to control the level of active auxins. Thus, the identification of auxin glycosyltransferases is of great significance for further understanding the auxin regulation. In this study, we biochemically screened the group L of Arabidopsis thaliana glycosyltransferase superfamily for enzymatic activity toward auxins. UGT74D1 was identified to be a novel auxin glycosyltransferase. Through HPLC and LC-MS analysis of reaction products in vitro by testing eight substrates including auxins and other compounds, we found that UGT74D1 had a strong glucosylating activity toward indole-3-butyric acid [IBA], indole-3-propionic acid [IPA], indole-3-acetic acid [IAA] and naphthaleneacetic acid [NAA], catalyzing them to form corresponding glucose esters. Biochemical characterization showed that this enzyme had a maximum activity in HEPES buffer at pH 6.0 and 37°C. In addition, the enzymatic activity analysis of crude protein and the IBA metabolite analysis from transgenic Arabidopsis plants overexpressing UGT74D1 gene were also carried out. Experimental results indicated that over-production of the UGT74D1 in plants indeed led to increased level of the glucose conjugate of IBA. Moreover, UGT74D1 overexpression lines displayed curling leaf phenotype, suggesting a physiological role of UGT74D1 in affecting the activity of auxins. Our current data provide a new target gene for further genetic studies to understand the auxin regulation by glycosylation in plants.
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