Section of Cell and Developmental Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, California 92093.
Plant Cell. 2010 Jan;22(1):249-59. doi: 10.1105/tpc.109.072066. Epub 2010 Jan 12.
To discover genes involved in nitric oxide (NO) metabolism, a genetic screen was employed to identify mutants defective in NO accumulation after treatment with the physiological inducer hydrogen peroxide. In wild-type Arabidopsis thaliana plants, NO levels increase eightfold in roots after H(2)O(2) treatment for 30 min. A mutant defective in H(2)O(2)-induced NO accumulation was identified, and the corresponding mutation was mapped to the prohibitin gene PHB3, converting the highly conserved Gly-37 to an Asp in the protein's SPFH domain. This point mutant and a T-DNA insertion mutant were examined for other NO-related phenotypes. Both mutants were defective in abscisic acid-induced NO accumulation and stomatal closure and in auxin-induced lateral root formation. Both mutants were less sensitive to salt stress, showing no increase in NO accumulation and less inhibition of primary root growth in response to NaCl treatment. In addition, light-induced NO accumulation was dramatically reduced in cotyledons. We found no evidence for impaired H(2)O(2) metabolism or signaling in the mutants as H(2)O(2) levels and H(2)O(2)-induced gene expression were unaffected by the mutations. These findings identify a component of the NO homeostasis system in plants and expand the function of prohibitin genes to include regulation of NO accumulation and NO-mediated responses.
为了发现参与一氧化氮(NO)代谢的基因,我们采用遗传筛选的方法来鉴定经生理诱导剂过氧化氢处理后 NO 积累缺陷的突变体。在野生型拟南芥中,H₂O₂处理 30 分钟后,根中的 NO 水平增加了 8 倍。我们鉴定到一个过氧化氢诱导的 NO 积累缺陷突变体,其相应的突变位于 PHB3 基因(即阻遏素基因),导致蛋白 SPFH 结构域中高度保守的甘氨酸 37 突变为天冬氨酸。该点突变体和 T-DNA 插入突变体被进一步用于研究其他与 NO 相关的表型。这两个突变体均不能积累 ABA 诱导的 NO,且气孔不能关闭,也不能形成生长素诱导的侧根。这两个突变体对盐胁迫的敏感性降低,NaCl 处理后 NO 积累没有增加,主根生长也没有受到抑制。此外,光诱导的 NO 积累在子叶中明显减少。我们没有发现突变体中 H₂O₂代谢或信号转导受损的证据,因为 H₂O₂水平及其诱导的基因表达不受突变的影响。这些发现鉴定了植物中 NO 动态平衡系统的一个组成部分,并扩展了阻遏素基因的功能,使其包括调节 NO 积累和 NO 介导的反应。