National Key Laboratory of Crop Genetic Improvement and National Center of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, China.
Plant Physiol. 2010 Feb;152(2):876-90. doi: 10.1104/pp.109.149856. Epub 2009 Dec 9.
Mitogen-activated protein kinase (MAPK) cascades have been identified in various signaling pathways involved in plant development and stress responses. We identified a drought-hypersensitive mutant (drought-hypersensitive mutant1 [dsm1]) of a putative MAPK kinase kinase (MAPKKK) gene in rice (Oryza sativa). Two allelic dsm1 mutants were more sensitive than wild-type plants to drought stress at both seedling and panicle development stages. The dsm1 mutants lost water more rapidly than wild-type plants under drought stress, which was in agreement with the increased drought-sensitivity phenotype of the mutant plants. DSM1-RNA interference lines were also hypersensitive to drought stress. The predicted DSM1 protein belongs to a B3 subgroup of plant Raf-like MAPKKKs and was localized in the nucleus. By real-time PCR analysis, the DSM1 gene was induced by salt, drought, and abscisic acid, but not by cold. Microarray analysis revealed that two peroxidase (POX) genes, POX22.3 and POX8.1, were sharply down-regulated compared to wild type, suggesting that DSM1 may be involved in reactive oxygen species (ROS) signaling. Peroxidase activity, electrolyte leakage, chlorophyll content, and 3,3'-diaminobenzidine staining revealed that the dsm1 mutant was more sensitive to oxidative stress due to an increase in ROS damage caused by the reduced POX activity. Overexpression of DSM1 in rice increased the tolerance to dehydration stress at the seedling stage. Together, these results suggest that DSM1 might be a novel MAPKKK functioning as an early signaling component in regulating responses to drought stress by regulating scavenging of ROS in rice.
植物发育和应激反应涉及的各种信号通路中已经鉴定出有丝分裂原激活的蛋白激酶(MAPK)级联。我们在水稻(Oryza sativa)中鉴定出一个假定的丝裂原激活蛋白激酶激酶激酶(MAPKKK)基因的干旱敏感突变体(干旱敏感突变体 1[dsm1])。两个等位基因的 dsm1 突变体在幼苗和穗发育阶段对干旱胁迫比野生型植物更敏感。在干旱胁迫下,dsm1 突变体比野生型植物更快地失去水分,这与突变体植物增加的干旱敏感性表型一致。DSM1-RNA 干扰系也对干旱胁迫敏感。预测的 DSM1 蛋白属于植物 Raf 样 MAPKKKs 的 B3 亚群,定位于细胞核中。通过实时 PCR 分析,DSM1 基因被盐、干旱和脱落酸诱导,但不受冷诱导。微阵列分析显示,与野生型相比,两个过氧化物酶(POX)基因 POX22.3 和 POX8.1 被明显下调,表明 DSM1 可能参与活性氧(ROS)信号转导。过氧化物酶活性、电解质渗漏、叶绿素含量和 3,3'-二氨基联苯胺染色表明,由于 POX 活性降低导致 ROS 损伤增加,dsm1 突变体对氧化应激更敏感。在水稻中过表达 DSM1 可提高幼苗期脱水胁迫的耐受性。综上所述,这些结果表明,DSM1 可能是一种新型的 MAPKKK,通过调节 ROS 的清除来作为早期信号成分调节水稻对干旱胁迫的响应。