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Cloning and expression analysis of BcOPR3 gene in non-heading Chinese cabbage
文锴 王远 胡蝶 袁敬平 侯喜林 李英
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, 江苏 南京 210095
WEN Kai WANG Yuan HU Die YUAN Jingping HOU Xilin LI Ying
State Key Laboratory of Crop Genetics and Germplasm Enhancement/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China, Ministry of Agriculture/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
Brassica campestris ssp. chinensis MakinoBcOPR3clonesubcellular locationRT-qPCR
[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BcOPR3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]BcOPR3在2个不结球白菜品种中的序列一致,其BcOPR3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BcOPR3定位在细胞膜上;实时荧光定量PCR(RT-qPCR)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BcOPR3基因表达。霜霉病菌侵染下,BcOPR3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BcOPR3,通过表达分析发现,BcOPR3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BcOPR3基因的高表达是其具有更高抗性的主要原因。
[Objectives]The 12-oxo-phytodienoic acid reductase(OPR)is a key enzyme in the biosynthesis pathway of jasmonic acid,which catalyzes the reduction of 12-oxo-phytodienoic acid to produce jasmonic acid compounds,and it is widely involved in the process of plant growth and defense system. Cloning and characterization of BcOPR3 from non-heading Chinese cabbage is significant for researching its effect on the responses of signal molecules and abiotic stress.[Methods]To explore the function of the OPR3 gene in non-heading Chinese cabbage,BcOPR3 was cloned and bioinformatics analysis was conducted. The sub-cellular localization was researched. The expression level of BcOPR3 was analyzed under Hyaloperonospora parasitica,abscissic acid(ABA),jasmonic acid(JA),salicylic acid(SA),mechanical injury,hypothermia and heat shock stress treatments.[Results]The sequence of BcOPR3 was consistent in two kinds of non-heading Chinese cabbage. The homology between Brassica campestris ssp.chinensis Makino and Brassica napus was up to 98%,based on the close relationship. BcOPR3 gene was verified to be located in the cytoplasm. RT-qPCR analysis showed that Hyaloperonospora parasitica,ABA,JA,SA,mechanical injury,hypothermia and heat shock stress induced the expression of BcOPR3. The BcOPR3 gene had a different expression between resistant line ‘Suzhouqing’ and susceptible inbred line ‘Aijiaohuang’ ,and it had a higher expression on the ‘Suzhouqing’ than the ‘Aijiaohuang’ under downy mildew infection.[Conclusions]The BcOPR3 gene may co-respond to abiotic stress and the gene plays an important role in the cytoplasm. The high transcription level of BcOPR3 gene is an important reason for the resistance of disease resistant varieties to downy mildew.


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更新日期/Last Update: 2017-09-23