牡丹PoCCS1基因的克隆与表达分析  

Cloning and Expression Analysis of PoCCS1 Gene in Tree Peony

作　　者：唐欣 周天华[1] 鲁仪增 张利 范志斌 李志祺 袁思怡 TANG Xin;ZHOU Tian-hua;LU Yi-zeng;ZHANG Li;FAN Zhi-bin;LI Zhi-qi;YUAN Si-yi(College of Agricultural and Biological Engineering,Heze Uninversity,Heze 274015,Shandong,China;Shandong Provincial Center of Forest and Grass Germplasm Resources,Jinan 250102,Shandong,China)

机构地区：[1]菏泽学院农业与生物工程学院,山东菏泽274015 [2]山东省林草种质资源中心,山东济南250102

出　　处：《西北林学院学报》2023年第2期84-91,共8页Journal of Northwest Forestry University

基　　金：菏泽学院博士基金(XY16BS34);菏泽学院培育项目(XY18PY05);菏泽学院牡丹研究院培育项目(PR15);山东省高等学校青创科技支持计划(鲁教人字〔2019〕6号);山东省自然科学基金青年项目(ZR2020QC166)。

摘　　要：铜/锌超氧化物歧化酶铜伴侣基因(copper chaperone for superoxide dismutase,CCS)负责将铜离子转运至Cu/Zn-SOD,从而激活SOD酶活性,参与植物活性氧清除,在植物抗逆中发挥重要作用。研究旨在克隆牡丹PoCCS 1基因,并对基因序列特征、组织表达模式、盐胁迫和干旱胁迫下的表达模式、不同牡丹品种氧化胁迫下的表达模式进行分析,为深入研究PoCCS 1在牡丹非生物胁迫响应中的功能奠定基础。利用RT-PCR技术克隆‘凤丹’PoCCS 1基因(GenBank登录号:MZ574405),利用生物信息学方法分析PoCCS1序列特征,利用实时荧光定量PCR(qRT-PCR)分析PoCCS 1的表达模式。结果表明,PoCCS 1基因编码区全长为996 bp,编码蛋白含331个氨基酸,相对分子量为34.98 ku,包含植物CCS蛋白的3个典型结构域,进化分析表明PoCCS1与多种植物的CCS同源性较高,且与葡萄VvCCS亲缘关系最近;PoCCS 1在‘凤丹’的根、茎和叶中表达水平相近,‘凤丹’盐胁迫8 h后,PoCCS 1在叶片和根中的表达受到显著诱导,干旱胁迫8 h后,PoCCS 1在叶片中的表达下调,根中表达无显著变化;4个抗氧化能力不同的牡丹品种‘鲁荷红’‘凤丹’‘香玉’和‘乌云集盛’氧化胁迫处理后PoCCS 1表达模式差异显著,随处理时间增加,在抗氧化能力强的‘乌云集盛’和‘香玉’中PoCCS 1的表达显著上调并在处理4 h后保持较高水平,在抗氧化能力较弱的‘凤丹’中表达稳定,在抗氧化能力最弱的‘鲁荷红’中先升后降并在处理4 h后显著下调。综上,PoCCS 1基因编码蛋白具有植物CCS的完整典型结构,可能参与牡丹响应盐胁迫和干旱胁迫,并可能与不同牡丹品种抗氧化能力差异相关。Copper(Cu)chaperone for superoxide dismutase(CCS)can activate Cu/Zn-SODs through delivering copper them,which plays important roles in reactive oxygen species scavenging and plant resistance.This study was conducted to clone PoCCS 1 gene from Paeonia ostii and analyze the gene sequence characteristics,tissue expression patterns,expression profiles under salt stress and drought stress,and expression patterns under oxidative stress of different peony varieties,so as to lay a foundation for further study on the function of PoCCS 1 in abiotic stress response in tree peony.PoCCS 1 gene(GenBank accession number:MZ574405)was cloned from'Feng Dan'by RT-PCR,PoCCS1 sequence characteristics were analyzed by bioinformatics,and the expression patterns of the PoCCS 1 were analyzed by real-time fluorescence quantitative PCR(qRT-PCR).The results showed that the full length of coding region of PoCCS 1 was 996 bp,which encoded protein contained 331 amino acids.The relative molecular weight of PoCCS1 was 34.98 ku,which contained 3 typical domains of plant CCS.Phylogenetic analysis indicated that PoCCS1 was highly homologous with many other plants and most closely related to VvCCS.The expression level of PoCCS 1 was similar in the leaves,stems,and roots of'Feng Dan'.After 8 hours under salt stress,the expression of PoCCS 1 in the leaves and roots was significantly induced.After 8 hours under drought stress,the expression of PoCCS 1 in the leaves were significantly down-regulated,while the expression in roots had no significant change.The expression patterns of PoCCS 1 under oxidative stress in four tree peony varieties'Lu He Hong','Feng Dan','Xiang Yu'and'Wu Yun Ji Sheng'with different antioxidant capacities were significantly different.With time of oxidative stress treatment increasing,PoCCS 1 expression in'Xiang Yu'and'Wu Yun Ji Sheng'with strong antioxidant capacity was significantly up-regulated,and maintained higher level after treatment for 4 h.The expression level of PoCCS 1 in'Feng Dan'with weak antioxidant capacity was stab

关 键 词：牡丹 PoCCS 1 克隆 非生物胁迫 表达分析 

分 类 号：S685.11[农业科学—观赏园艺]