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园艺学报 ›› 2015, Vol. 42 ›› Issue (12): 2497-2504.doi: 10.16420/j.issn.0513-353x.2015-0098

• 研究报告 • 上一篇    下一篇

枳橙CPMAX2基因的克隆、载体构建及其表达研究

袁飞荣1,2,袁耀明2,3,李智鑫2,3,Alessandra Gentile4,邓子牛5,*   

  1. 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 University,Dipartimento Diortofloroarboricolturae Tecnologie Agroalimentari,Catania 95123,Italy;5国家柑橘改良中心长沙分中心,长沙 410128
  • 出版日期:2015-12-25 发布日期:2015-12-25
  • 基金资助:

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

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

YUAN Fei-rong1,2,YUAN Yao-ming2,3,LI Zhi-xin2,3,Alessandra Gentile4,and DENG Zi-niu5,*   

  1. 1Department of Landscape Architecture,Wuhan Bioengineering Institute,Wuhan 430415,China;2 Center of Applied Biotechnology,Wuhan Bioengineering Institute,Wuhan 430415,China;3 College of Life Science and Technology,Wuhan Bioengineering Institute,Wuhan 430415,China;4 Dipartimento Diortofloroarboricolturae Tecnologie Agroalimentari,Catania University,Catania 95123,Italy;5 National Center of Citrus Improvement,Changsha Subcenter,Changsha 410128,China
  • Online:2015-12-25 Published:2015-12-25

摘要:

转rolABC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rolABC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rolABC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rolABC基因枳橙莲座型分枝生长中扮演重要角色。

关键词: 柑橘, 独脚金内酯, CPMAX2, 腋芽萌发

Abstract:

Transgenic citrange lines with rolABC 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 cDNA 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-pCAMBIA1301 for further study. The results showed that the cDNA 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 arehighly 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 rolABC genes.

Key words: citrus, strigolactone, CPMAX2 gene, axillary bud outgrowth

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