Tanaka Keita, Hayashi Ken-ichiro, Natsume Masahiro, Kamiya Yuji, Sakakibara Hitoshi, Kawaide Hiroshi, Kasahara Hiroyuki
United Graduate School of Agricultural Science, Tokyo University of Agriculture & Technology, Tokyo, 183-8509 Japan.
Plant Cell Physiol. 2014 Jan;55(1):218-28. doi: 10.1093/pcp/pct173. Epub 2013 Nov 26.
IAA is a naturally occurring auxin that plays a crucial role in the regulation of plant growth and development. The endogenous concentration of IAA is spatiotemporally regulated by biosynthesis, transport and its inactivation in plants. Previous studies have shown that the metabolism of IAA to 2-oxindole-3-acetic acid (OxIAA) and OxIAA-glucoside (OxIAA-Glc) may play an important role in IAA homeostasis, but the genes involved in this metabolic pathway are still unknown. In this study, we show that UGT74D1 catalyzes the glucosylation of OxIAA in Arabidopsis. By screening yeasts transformed with Arabidopsis UDP-glycosyltransferase (UGT) genes, we found that OxIAA-Glc accumulates in the culture media of yeasts expressing UGT74D1 in the presence of OxIAA. Further, we showed that UGT74D1 expressed in Escherichia coli converts OxIAA to OxIAA-Glc. The endogenous concentration of OxIAA-Glc decreased by 85% while that of OxIAA increased 2.5-fold in ugt74d1-deficient mutants, indicating the major role of UGT74D1 in OxIAA metabolism. Moreover, the induction of UGT74D1 markedly increased the level of OxIAA-Glc and loss of root gravitropism. These results indicate that UGT74D1 catalyzes a committed step in the OxIAA-dependent IAA metabolic pathway in Arabidopsis.
吲哚-3-乙酸(IAA)是一种天然存在的生长素,在植物生长和发育的调控中起着关键作用。IAA的内源浓度在植物体内通过生物合成、运输及其失活进行时空调节。先前的研究表明,IAA代谢为2-氧代吲哚-3-乙酸(OxIAA)和OxIAA-葡萄糖苷(OxIAA-Glc)可能在IAA稳态中发挥重要作用,但参与该代谢途径的基因仍不清楚。在本研究中,我们表明UGT74D1催化拟南芥中OxIAA的糖基化反应。通过筛选用拟南芥尿苷二磷酸糖基转移酶(UGT)基因转化的酵母,我们发现,在存在OxIAA的情况下,OxIAA-Glc在表达UGT74D1的酵母培养基中积累。此外,我们表明在大肠杆菌中表达的UGT74D1将OxIAA转化为OxIAA-Glc。在ugt74d1缺陷型突变体中,OxIAA-Glc的内源浓度降低了85%,而OxIAA的浓度增加了2.5倍,这表明UGT74D1在OxIAA代谢中起主要作用。此外,UGT74D1的诱导显著增加了OxIAA-Glc的水平并导致根失去向重力性。这些结果表明,UGT74D1催化拟南芥中依赖OxIAA的IAA代谢途径中的一个关键步骤。