枣树ZjSOD1基因参与非生物胁迫响应的研究  

Studies on the role of ZjSOD1 gene from Ziziphus jujuba in response to abiotic stresses

作　　者：聂园军 郭慧娜 张春芬 肖蓉 邓舒 李倩 曹秋芬 NIE Yuanjun;GUO Huina;ZHANG Chunfen;XIAO Rong;DENG Shu;LI Qian;CAO Qiufen(College of Agricultural Economics&Management,Shanxi Agricultural University,Taiyuan 030006,Shanxi,China;Institute of Pomology,Shanxi Agricultural University,Taigu 030815,Shanxi,China;College of Life Sciences,Shanxi Agricultural University,Taiyuan 030031,Shanxi,China)

机构地区：[1]山西农业大学农业经济管理学院,太原030006 [2]山西农业大学果树研究所,山西太谷030815 [3]山西农业大学生命科学学院,太原030031

出　　处：《果树学报》2020年第9期1294-1304,共11页Journal of Fruit Science

基　　金：国家自然科学基金(31372033);国家基础研究计划项目(2012FY110100-5);山西省科技成果转化引导专项(201904D131039);山西省农业科学院攻关项目(YGG1432)。

摘　　要：【目的】克隆1个枣树超氧化物歧化酶基因ZjSOD1,并对其功能进行研究,为其在枣树抗逆基因工程改良中的利用奠定基础。【方法】采用PCR克隆ZjSOD1 cDNA序列,运用实时定量RT-PCR方法研究其在高盐和PEG6000胁迫下转录水平的变化。构建ZjSOD1过量表达载体转化拟南芥,研究其在拟南芥中响应非生物胁迫应答的功能,测定氧化酶活性及生理指标。【结果】ZjSOD1基因cDNA序列开放阅读框全长为699 bp,编码232个氨基酸,理论等电点为8.59,属于Fe-SOD家族成员。在转录水平上,ZjSOD1明显受NaCl和PEG6000诱导。在拟南芥中过量表达ZjSOD1基因导致植株对干旱、高盐更加敏感,但对H2O2耐受性显著增强。在NaCl、PEG6000和H2O2胁迫条件下,过量表达Zj-SOD1转基因植株中SOD、CAT和POD酶活性、脯氨酸和丙二醛含量、电解质渗透率均显著发生改变。实时定量RTPCR结果显示参与活性氧(ROS)、高盐和干旱胁迫相关信号通路的基因在ZjSOD1转基因株系中表达量发生明显改变。【结论】ZjSOD1在植物生长发育过程中参与氧化、干旱、高盐胁迫应答。【Objective】Superoxide dismutases(SODs)are involved in protecting plants against diverse biotic and abiotic stresses.In this study,a novel Fe-SOD gene,named ZjSOD1,was cloned from Ziziphus jujuba Mill.Hupingzao and its function was studied.The expression of ZjSOD1 of jujube plantlets generated from tissue culture was studied under different abiotic stress treatments.【Methods】The ORF sequences of the ZjSOD1 cDNA was cloned by polymerase chain reaction(PCR).The expression of the ZjSOD1 under salt,and drought stresses was analyzed by quantitative RT-PCR.To characterize the function of the ZjSOD1 gene in plant development,coding sequence of ZjSOD1 was inserted into the plant expression vectors PEZR(K)-LNY driven by CaMV 35S promoter and introduced into Arabidopsis.Subsequently,seeds of the wild type and homozygotes of T3 generation of the ZjSOD1 overexpression transgenic lines of Arabidopsis were sowed on MS agar media with or without NaCl,PEG6000 and H2O2,respectively.The seed germination rates,the cotyledon greening rate and the growth rates of the primary roots of the wild type and ZjSOD1 lines were monitored under different abiotic stresses.The levels of proline,malondialdehyde(MDA),electrolyte leakage and the specific activities of various antioxidant enzymes were detected in the leaf tissues for evaluating the extent of cellular damage of both the ZjSOD1 transgenic lines and the wild type during salt,PEG6000 and H2O2 treatments.In addition,we analyzed the expression levels of antioxidant enzymes(APX1,GPX3 and CAT1)in the ROS scavenging system,drought/high salinity-responsive genes(RD29A and ERD15)and SOS2(involved in SOS signaling pathway)were employed as markers for monitoring ROS signaling and stress-response pathways in Arabidopsis by quantitative RT-PCR.【Results】The ZjSOD1 contained a 699 bp of open reading frame(ORF)encoding a protein of 232 amino acids(including 22 acidic amino acids and 24 basic amino acids)with a calculated molecular mass of 25.5971 ku and an isoelectric point of 8.59.Zj-SOD1

关 键 词：枣树 ZjSOD1 渗透/高盐/H2O2胁迫 幼苗发育 

分 类 号：S665.1[农业科学—果树学]