不结球白菜BcOPR3基因的克隆与功能分析

发布时间：2018-03-19 01:23

本文选题：不结球白菜　切入点：BcOPR　出处：《南京农业大学学报》2017年05期 　论文类型：期刊论文

【摘要】：[目的]12-氧-植物二烯酸还原酶(12-oxo-phytodienoic acid reductase,OPR)是茉莉酸生物合成途径的关键酶,催化12-氧-植物二烯(OPDA)还原反应生成茉莉酸化合物,广泛参与植物生长过程中的防御系统。本文旨在克隆和研究不结球白菜Bc OPR3基因,研究其对信号分子和非生物胁迫应答的影响。[方法]对不结球白菜OPR3基因进行克隆、生物信息分析、亚细胞定位,并对其在霜霉病菌、脱落酸(ABA)、茉莉酸(JA)、水杨酸(SA)、机械伤害、低温和热激胁迫下的表达水平进行了分析。[结果]Bc OPR3在2个不结球白菜品种中的序列一致,其Bc OPR3蛋白与同科油菜(Brassica napus)的同源性高达98%,进化关系与之最相近;亚细胞定位结果显示Bc OPR3定位在细胞膜上;实时荧光定量PCR(RT-q PCR)检测结果表明,不结球白菜霜霉病菌、ABA、JA、SA、机械伤害、低温和热激胁迫均能诱导Bc OPR3基因表达。霜霉病菌侵染下,Bc OPR3在抗性自交系‘苏州青’和感病自交系‘矮脚黄’中存在差异表达,并且在抗病自交系中表达量较高。[结论]从不结球白菜克隆得到的Bc OPR3,通过表达分析发现,Bc OPR3可能存在对非生物胁迫的共响应,且在不结球白菜中该基因是在细胞膜上起作用。在非生物胁迫下,基因具有不同的应答模式,抗性自交系Bc OPR3基因的高表达是其具有更高抗性的主要原因。
[Abstract]:[objective] 12-oxo-phytodienoic acid reductases (12-oxo-phytodienoic acid reductases), which catalyze the reduction of 12-oxo-phytodienoic acid reductases to jasmonate compounds, are the key enzymes in the biosynthesis of jasmonic acid. The purpose of this paper is to clone and study the Bc OPR3 gene of non-heading cabbage and its effect on signal molecule and abiotic stress response. [methods] the OPR3 gene of non-heading Chinese cabbage was cloned. Biological information analysis, subcellular localization, and mechanical injury in downy mildew, abscisic acid abscisic acid, jasmonic acid, salicylate, SAA, The expression level of BC OPR3 was analyzed under low temperature and heat shock stress. [results] the sequence of BC OPR3 in two non-heading Chinese cabbage cultivars was the same, and the similarity of BC OPR3 protein to Brassica napus was 98, and the evolutionary relationship was the most similar. The results of subcellular localization showed that BC OPR3 was located on the cell membrane, and the results of real-time fluorescence quantitative PCR(RT-q PCR showed that the non-heading Chinese cabbage downy mildew strain ABAJA JASAA was mechanical injury. The expression of BC OPR3 gene was induced by low temperature stress and heat shock stress. Bc OPR3 was differentially expressed in resistant inbred lines' Suzhou green 'and susceptible inbred line' dwarf foot yellow 'under downy mildew infection. The expression level of Bc OPR3 was higher in resistant inbred lines. [conclusion] Bc-OPR3, which was cloned from non-heading Chinese cabbage, was found to have co-response to abiotic stress by expression analysis. Under abiotic stress, the gene had different response patterns, and the high expression of BC OPR3 gene in resistant inbred lines was the main reason for its higher resistance.
【作者单位】：南京农业大学作物遗传与种质创新国家重点实验室/农业部华东地区园艺作物生物学与种质创新重点实验室/南京农业大学园艺学院;
【基金】：国家自然科学基金项目(31272173,31471886)
【分类号】：S634.3

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