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非特异性磷脂酶C(NPC,nonspecific phospholipase C)水解磷脂酰胆碱或磷脂酰乙醇胺产生第二信使二酰甘油(DAG),NPC类基因在植物处于干旱、高盐、低磷等非生物胁迫条件下时发挥重要的作用,并参与脱落酸(ABA)、油菜素内酯(BL)等激素信号传导途径。本研究以谷子品种龙谷25为试验材料,通过序列比对,克隆到一个新的NPC类基因,命名为Si NPC4。该基因被定位在谷子第8条染色体上,编码区全长2877 bp,由3个外显子和2个内含子组成,编码512个氨基酸,蛋白分子量为56.77 k D。系统进化树分析表明该基因位于NPC基因家族第3亚族,保守结构域分析表明该蛋白含有保守的磷脂酶结构域和4个基序。亚细胞定位结果显示,Si NPC4蛋白被定位在细胞质、细胞膜、细胞核中。基因表达谱分析结果表明,该基因主要在谷子根部表达,并受干旱、盐、低温、黑暗、ABA、BL、赤霉素(GA)和茉莉酸甲酯(Me JA)等的诱导表达。将Si NPC4基因转入拟南芥中,转基因拟南芥与野生型拟南芥相比对ABA和BL激素的敏感性降低,推测该基因可能作为负调控因子参与植物对ABA和BL激素的响应过程。在干旱和高盐处理下,过表达植株与野生型植株长势没有明显差异。
Non-specific phospholipase C (NPC) hydrolyzes phosphatidylcholine or phosphatidylethanolamine to generate second messenger diacylglycerol (DAG). The NPC genes are under conditions of abiotic, high salt, Play an important role under the next, and participate in hormone ABA, brassinosteroid (BL) and other hormone signaling pathways. In this study, the millet variety Longgu 25 was used as experimental material, and a new NPC gene was cloned by sequence alignment and named as Si NPC4. The gene is located on chromosome 8 of the millet. The coding region is 2877 bp in length. It consists of 3 exons and 2 introns and encodes a protein of 512 amino acids with a molecular mass of 56.77 kD. Phylogenetic tree analysis showed that this gene was located in the third subfamily of NPC gene family. The analysis of conserved domain showed that the protein contains conserved phospholipase domain and four motifs. Subcellular localization results show that Si NPC4 protein is localized in the cytoplasm, cell membrane and nucleus. The results of gene expression profiling showed that the gene was mainly expressed in the roots of millet and was induced by drought, salt, hypothermia, darkness, ABA, BL, GA and Me JA. The Si NPC4 gene was transferred into Arabidopsis and the sensitivity of transgenic Arabidopsis plants to ABA and BL hormone decreased compared with that of wild-type Arabidopsis thaliana, suggesting that this gene may be involved in plant responses to ABA and BL hormones as a negative regulator process. Under drought and high salinity treatments, there was no significant difference in the growth of over-expressing and wild-type plants.