枳橙CPMAX2基因的克隆、载体构建及其表达研究  

Cloning,Constructing the Over-expression Vector and Expression Analysis of CPMAX2 in Transgenic Citrange with rolABC Genes

作　　者：袁飞荣[1,2] 袁耀明[2,3] 李智鑫[2,3] Alessandra Gentile 邓子牛[5] 

机构地区：[1]武汉生物工程学院园林系,武汉410415 [2]武汉生物工程学院应用生物技术研究中心,武汉430415 [3]武汉生物工程学院生命科学与技术学院,武汉430415 [4]Catania Univeralty,Dipartimento Diortofloroarboricoltuae Tecnologle Agroallmentarl,Catania 95123,Italy [5]国家柑橘改良中心长沙分中心,长沙410128

出　　处：《园艺学报》2015年第12期2497-2504,共8页Acta Horticulturae Sinica

基　　金：国家自然科学基金青年基金项目(31301750);国家‘863’计划项目(2011AA100205)

摘　　要：转rol ABC基因枳橙莲座状分枝、矮化性状突出。为探索其侧枝生长的植物激素调控机制,通过RT-PCR法从枳橙中克隆到独脚金内酯信号转导关键元件CPMAX2基因,分析了该基因在转rol ABC基因枳橙腋生嫩叶中的转录表达,构建了CPMAX2植物过表达载体。研究表明:CPMAX2与甜橙中同源基因一致性为99.66%,编码694个氨基酸,具有一个典型的F-box蛋白结构域和两个LRR重复区,与甜橙MAX2氨基酸序列一致性为99.14%。CPMAX2在转rol ABC基因枳橙3个株系腋生嫩叶中的转录表达均比野生型显著下调。试验表明CPMAX2在转rol ABC基因枳橙莲座型分枝生长中扮演重要角色。Transgenic citrange lines with rol ABC genes behave resset branching and extreme dwarfing. To explore the regulatory mechanism of plant hormones in axillary shoot growth of transgenic citrange,a full length c DNA of CPMAX2 was cloned from citrange[Citrus sinensis（L.）Osb × Poncirus trifoliate（L.）Raf.] by RT-PCR in this study. The expression of CPMAX2 was detected in axillary tender leaves of 3 transgenic citrange lines and the wild type;Additionally,we constructed an over-expression vector CPMAX2-p CAMBIA1301 for further study. The results showed that the c DNA sequence and its putative peptide sequence shared 99.66% and 99.14% of identity with its citrus sinensis ortholog MAX2. The deduced amino acid sequence contained putatively one F-box and two LRR repeat domains that are highly conserved in MAX2 genes. CPMAX2 was obviously down-expressed in tender leaves of 3 transgenic citrange lines compared with the wild type. The results suggest CPMAX2 play an important role in regulating the rosette shoot growth in transgenic citrange with rol ABC genes.

关 键 词：柑橘 独脚金内酯 CPMAX2 腋芽萌发 

分 类 号：S666[农业科学—果树学]