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植物研究 ›› 2016, Vol. 36 ›› Issue (6): 902-908.doi: 10.7525/j.issn.1673-5102.2016.06.014

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苹果生长素输入载体MdAUX1的基因克隆及在拟南芥和烟草中的遗传转化

安建平, 王小非, 刘鑫, 李浩浩, 由春香, 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, 泰安 271018
  • 收稿日期:2016-09-05 出版日期:2016-11-15 发布日期:2016-11-16
  • 通讯作者: 郝玉金,E-mail:haoyujin@sdau.edu.cn E-mail:haoyujin@sdau.edu.cn
  • 作者简介:安建平(1990-),男,硕士研究生,主要从事果树生物技术的研究。
  • 基金资助:
    国家自然科学基金(31430074);山东省农业工程和产业体系(SDAIT-03-033-03)

Molecular Cloning and Genetic Transformation Analysis of an Apple Auxin Influx Transporter Gene MdAUX1 in Arabidopsis thaliana and Tobacco

AN Jian-Ping, WANG Xiao-Fei, LIU Xin, LI Hao-Hao, YOU Chun-Xiang, HAO Yu-Jin   

  1. College of Horticulture Science and Engineering, Shandong Agricultural University, State Key Laboratory of Crop Biology, Tai'an 271018
  • Received:2016-09-05 Online:2016-11-15 Published:2016-11-16
  • Supported by:
    The National Natural Science Foundation of China(31430074);Shandong Agricultural Engineering and industrial System(SDAIT-03-033-03)

摘要: 生长素的极性运输对调节植物生长发育起重要作用。生长素转运蛋白调控生长素的极性运输。本研究从‘嘎啦’(Malus×domestic‘Royal Gala’)苹果中克隆了生长素输入载体基因MdAUX1(基因序列号:MDP0000384437)。序列分析表明,该基因包含长为1 443 bp完整的开放阅读框,编码含有480个氨基酸的蛋白。氨基酸序列比对和系统进化树分析表明,AUX1在不同物种间具有高度的序列保守性,其中,苹果MdAUX1与欧洲甜樱桃PaAUX1亲缘关系最近。半定量PCR分析显示,MdAUX1在苹果的茎中表达量较高。通过农杆菌介导的遗传转化获得转MdAUX1基因的拟南芥和烟草。转基因拟南芥根系表型观察结果显示,MdAUX1基因在拟南芥中异位表达后,会抑制主根伸长,促进侧根形成;在外源NAA处理下,转基因拟南芥对主根伸长的抑制作用得到部分恢复。在烟草中过量表达MdAUX1基因能够显著促进植株地上部分生长。以上结果说明苹果MdAUX1基因在调节植物生长发育中发挥重要作用。

关键词: 苹果, 生长素输入载体, MdAUX1, 遗传转化

Abstract: Auxin has been identified to play critical roles in regulating plant growth and development. The polar transport of auxin is regulated by auxin transporters. In this study, an auxin influx transporter gene named MdAUX1 was cloned from Malus×domestic‘Royal Gala’. Sequence analysis showed that the length of MdAUX1 gene was 1 443 bp, which encoded 480 amino acids. Amino acid sequence alignment and phylogenetic tree analysis showed that the AUX1 had a high sequence conservation among different species, and apple MdAUX1 exhibited the highest sequence similarity with Prunus avium PaAUX1. RT-PCR analysis demonstrated the MdAUX1 gene was mainly expressed in stem. We generated transgenic Arabidopsis and tobacco plants expressing the apple MdAUX1 by Agrobacterium-mediated genetic transformation. Ectopic expression of MdAUX1 in Arabidopsis exhibited the phenotype of inhibition of primary root elongation and increase lateral root number. Furthermore, there was no significant inhibition on primary root elongation phenotype when treated with the NAA. Overexpressing MdAUX1 in tobacco could significantly promote the growth of aerial parts of plant. These results suggested that MdAUX1 played a vital role in plant growth and development.

Key words: apple, auxin influx transporters, MdAUX1, genetic transformation

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