荔枝古树胚性愈伤组织LcCu/Zn-SOD3基因启动子的克隆及功能验证  

Cloning and Functional Analysis of LcCu/Zn-SOD3 Promoter from Embryogenic Callus of the Ancient Litchi Tree

作　　者：练从龙[1] 卢秉国[1] 赖钟雄[1] 冯新[1] 林玉玲[1] 陈裕坤[1] 张梓浩[1] 

机构地区：[1]福建农林大学园艺植物生物工程研究所,福州350002

出　　处：《西北植物学报》2015年第1期16-22,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：福建省农业科技平台(2008N2001);国家科技支持计划项目(2007BAD07B01)

摘　　要：以荔枝古树"宋荔"胚性愈伤组织为材料,采用接头染色体步移法,分离获得LcCu/Zn-SOD3启动子片段长度为1 426bp,命名为ProLcCSD3(GenBank登录号:KF672186.1)。生物信息学分析表明,该启动子含有多个逆境应答元件、激素应答元件、胚乳特异表达元件,且可能受WRKY和MYB等转录因子调控。通过双酶切方法,以ProLcCSD3替换载体pCAMBIA1301上的CaMv35S启动子,构建了重组质粒p1301-proLcCSD3-GUS,并成功转化农杆菌EHA105和GV3101。注射烟草的瞬时表达分析表明,该启动子片段可以驱动下游报告基因表达。转化拟南芥分析表明,该启动子驱动的下游GUS可以在拟南芥的根、茎、叶中表达,且可响应NaCl、PEG-6000、ABA、MeJA和损伤等非生物胁迫。研究表明,荔枝古树LcCu/Zn-SOD3可能参与多种非生物胁迫应答和激素信号转导途径。In this experiment, the 1 426 bp promoter fragment of LcCu/Zn-SOD3 from embryogenic callus of the ancient litchi tree ＇Songli＇ which named ProLcCSD3（GenBank. KF672186.1） was cloned by means of adapter chromosome walking method. The bioinformaties analysis showed that ProLcCSD3 contains a number of environmental stress response elements,hormone response elements,endosperm expression elements and also might be regulation by MYB and WRKY transcription factors. A novel plant expression vector p1301-proLcCSD3-GUS was constructed with ProLcCSD3 replace 35S promoter in the pCAMBIA1301 vector by using the double-digested method and then transferred into Agrobacterium strain EHA105 and GV3101. Furthermore,transformation analysis showed that ProLcCSD3 has promoter activity by the infection of tobacco leaves and could drive the downstream reporter GUS gene expression in the roots, stems and leaves by Agrobacterium-mediated transformation of Arabidopsis. And stress treatments showed that Pro- LcCSD3 could response to some abiotic stresses, such as NaC1,PEG-6000,ABA, MeJA and wounding. Our results demonstrated that LcCu/Zn-SOD3 might response to abiotic stresses and participate in the signal transduction pathways of hormones in the ancient litchi tree.

关 键 词：荔枝古树 胚性愈伤组织 Cu/Zn-SOD3 启动子 功能验证 

分 类 号：Q785[生物学—分子生物学] Q786