Phospholipase Dα1 and Phosphatidic Acid Regulate NADPH Oxidase Activity and Production of Reactive Oxygen Species in ABA-Mediated Stomatal Closure in Arabidopsis
Phospholipase Dα1 and Phosphatidic Acid Regulate NADPH Oxidase Activity and Production of Reactive Oxygen Species in ABA-Mediated Stomatal Closure in Arabidopsis
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DOI：
10.1105/tpc.108.062992
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发表时间：
2009-08-01
期刊：
影响因子：
11.6
通讯作者：
Wang, Xuemin
中科院分区：
文献类型：
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作者：
Zhang, Yanyan;Zhu, Huiying;Wang, Xuemin
我们确定了磷脂酶 D α 1 (PLD α 1) 及其脂质产物磷脂酸 (PA) 在脱落酸 (ABA) 诱导拟南芥保卫细胞中活性氧 (ROS) 产生中的作用。 pld α 1 突变体未能在保卫细胞中响应 ABA 产生 ROS。 ABA 刺激野生型保卫细胞中的 NADPH 氧化酶活性，但不刺激 pld α 1 细胞中的 NADPH 氧化酶活性，而 PA 刺激两种基因型中的 NADPH 氧化酶活性。 PA 与重组拟南芥 NADPH 氧化酶 RbohD（呼吸爆发氧化酶同源物 D）和 RbohF 结合。鉴定了 PA 结合基序，RbohD 中 Arg 残基 149、150、156 和 157 的突变导致 PA 结合丧失和 RbohD 的 PA 激活丧失。表达非 PA 结合 RbohD 的 rbohD 突变体在 ABA 介导的 ROS 产生和气孔关闭中受到损害。此外，PLD α1 保卫细胞中 ABA 诱导的一氧化氮 (NO) 产生受到损害。 PA 与 ABI1 蛋白磷酸酶 2C 结合的破坏不会影响 ABA 诱导的 ROS 或 NO 的产生，但 PA-ABI1 相互作用是 ABA、H2O2 或 NO 诱导的气孔关闭所必需的。因此，PA 是连接保卫细胞中 ABA 信号网络中不同成分的中心脂质信号分子。
We determined the role of Phospholipase D alpha 1 (PLD alpha 1) and its lipid product phosphatidic acid (PA) in abscisic acid (ABA)-induced production of reactive oxygen species (ROS) in Arabidopsis thaliana guard cells. The pld alpha 1 mutant failed to produce ROS in guard cells in response to ABA. ABA stimulated NADPH oxidase activity in wild-type guard cells but not in pld alpha 1 cells, whereas PA stimulated NADPH oxidase activity in both genotypes. PA bound to recombinant Arabidopsis NADPH oxidase RbohD (respiratory burst oxidase homolog D) and RbohF. The PA binding motifs were identified, and mutation of the Arg residues 149, 150, 156, and 157 in RbohD resulted in the loss of PA binding and the loss of PA activation of RbohD. The rbohD mutant expressing non-PA-binding RbohD was compromised in ABA-mediated ROS production and stomatal closure. Furthermore, ABA-induced production of nitric oxide (NO) was impaired in pld alpha 1 guard cells. Disruption of PA binding to ABI1 protein phosphatase 2C did not affect ABA-induced production of ROS or NO, but the PA-ABI1 interaction was required for stomatal closure induced by ABA, H2O2, or NO. Thus, PA is as a central lipid signaling molecule that links different components in the ABA signaling network in guard cells.