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园艺学报 ›› 2018, Vol. 45 ›› Issue (6): 1041-1053.doi: 10.16420/j.issn.0513-353x.2017-0719

• 研究论文 • 上一篇    下一篇

苹果生长素阻遏蛋白基因MdIAA26的分子克隆与功能鉴定

韩朋良,刘肖娟,刘 鑫,董元花,胡大刚*,郝玉金*   

  1. 山东农业大学园艺科学与工程学院,作物生物学国家重点实验室,农业部黄淮地区园艺作物生物学与种质创制重点实验室,山东泰安 271018
  • 出版日期:2018-06-25 发布日期:2018-06-25
  • 基金资助:
    国家自然科学基金项目(31601728);山东省自然科学基金项目(ZR2016CQ13);山东农业大学杰出青年基金项目(564024);山东农业大学科技创新基金项目(24024)

Molecular Cloning and Functional Identification of Apple Auxin Repressor Protein Gene MdIAA26

HAN Pengliang,LIU Xiaojuan,LIU Xin,DONG Yuanhua,HU Dagang*,and HAO Yujin*   

  1. State Key Laboratory of Crop Biology,MOA Key Laboratory of Horticultural Crop Biology(Huanghuai Region)and Germplasm Innovation,College of Horticulture Science and Engineering,Shandong Agricultural University,Tai’an,Shandong 271018,China
  • Online:2018-06-25 Published:2018-06-25

摘要: 从‘皇家嘎拉’苹果(Malus × domestica Borkh.)中克隆了一个生长素阻遏蛋白家族基因(基因序列号MDP0000164095)。测序发现,该基因包含长为978 bp完整的开放阅读框,编码325个氨基酸。系统进化树分析表明,这一生长素阻遏蛋白与拟南芥IAA26同源序列相似性最高,因此将该基因命名为MdIAA26。利用PlantCare数据库进行基因启动子顺式作用元件预测分析发现,MdIAA26启动子序列中含有与脱落酸(ABA)、赤霉素(GA)、水杨酸(SA)及干旱信号相关的顺式作用元件。荧光定量PCR分析表明,MdIAA26在苹果的各组织中均有表达,在根和叶片中表达量相对较高;苹果组培苗中MdIAA26的表达明显受到ABA和IAA的诱导。MdIAA26过量表达的苹果愈伤组织在外源ABA处理下和MdIAA26异位表达拟南芥在外源ABA、PEG和IAA处理条件下,生长势均明显比野生型对照强,表明MdIAA26降低了对ABA和IAA的敏感性和对干旱的抗性。

关键词: 苹果, MdIAA26, AtIAA26, 生长素阻遏蛋白, 脱落酸

Abstract: An auxin repressor protein family gene(GenBank accession number:MDP0000164095)was cloned from‘Royal Gala’apple cultivar(Malus × domestica Borkh.). It contains complete open reading frame(ORF)of 978 bp in length and encodes 325 amino acids. Phylogenetic tree analysis showed that the deduced peptides exhibited the highest sequence similarity with AtIAA26(Arabidopsis),which was also indicated as an auxin repressor. Based on the result,we named this deduced gene as MdIAA26. In silico analysis suggested that the promoter sequence of MdIAA26 contained several typical cis-acting elements,including abscisic acid-,gibberellin-,salicylic acid- and drought-responsive elements by using PlantCare Databases. qPCR assays showed that MdIAA26 constitutively expressed in different tissues of apple,with higher expression levels in roots and leaves. The expression pattern of MdIAA26 in apple tissue culture seedling was noticeably induced by exogenous ABA and IAA. Further,the MdIAA26 overexpressed apple calli were treated with exogenous ABA and ectopical expressed Arabidopsis plants were treated with exogenous ABA,PEG and IAA. The growth potential of transgenic materials were significantly stronger than the wild type(WT,the control). All this suggests that transgenic materials were more insensitive to exogenous ABA and IAA and less resistant to drought than the control.

Key words: apple, MdIAA26, AtIAA26, auxin repressor protein, abscisic acid

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