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园艺学报 ›› 2019, Vol. 46 ›› Issue (10): 1989-1998.doi: 10.16420/j.issn.0513-353x.2019-0004

• 研究论文 • 上一篇    下一篇

甜瓜CmCRL1的克隆与表达分析

张焕欣,曹 娜,杨惠栋,李国权*,朱方红*   

  1. 江西省农业科学院园艺研究所,南昌 330200
  • 出版日期:2019-10-25 发布日期:2019-10-25
  • 基金资助:
    国家现代农业产业技术体系建设专项资金项目(CARS-25);公益性行业(农业)科研专项(201503110-05);江西省青年科学基金项目(20192BAB214017);江西省科技支撑计划项目(20141BBF60017);江西省农业科学院创新基金项目(20181CBS002)

Cloning and Expression Analysis of CmCRL1 in Melon

ZHANG Huanxin,CAO Na,YANG Huidong,LI Guoquan*,and ZHU Fanghong*   

  1. Institute of Horticulture,Jiangxi Academy of Agricultural Sciences,Nanchang 330200,China
  • Online:2019-10-25 Published:2019-10-25

摘要: 以甜瓜(Cucumis melo L.)品系‘L8’为试材,采用PCR方法克隆到CmCRL1(Crown Rootless1)基因,登录号为MELO3C012908,并通过Real-time PCR方法检测了CmCRL1的组织表达特异性和对低温、山梨醇、PEG6000、盐胁迫和淹水胁迫的响应模式。结果显示:CmCRL1含2个外显子和1个内含子,cDNA序列长度为996 bp,开放阅读框(ORF)长度为732 bp,编码243个氨基酸;CmCRL1蛋白中存在1个保守的LOB结构域,属于LBD/AS2(Lateral Organ Boundaries Domain/Asymmetric Leaves2)家族成员;多重比对分析发现CmCRL1与黄瓜Csa3G396920、西瓜Cla005992、拟南芥AtLBD16和AtLBD29、水稻OsCRL1、玉米ZmRTCS和ZmRTCL的LOB结构域高度保守,序列相似性分别为100%、99.02%、74.51%、86.27%、88.24%、89.22%和80.39%;组织表达特异性分析显示CmCRL1在根中优势表达,表达量分别是子叶、下胚轴、上胚轴、叶片和茎尖的178.39倍、7.54倍、26.95倍、65.19倍和75.86倍;CmCRL1能响应低温、山梨醇、PEG6000和盐等胁迫;此外,在淹水胁迫下甜瓜不定根发生过程中CmCRL1表达上调。

关键词: 甜瓜, CmCRL1, 基因克隆, 基因表达

Abstract: In the present study,CmCRL1(MELO3C012908)was cloned by PCR approach from‘L8’melon strain(Cucumis melo L.). The tissue-specific transcription pattern of CmCRL1 and responses to low temperature,sorbitol,PEG6000,salt and waterlogging stresses were analyzed by real-time PCR method. CmCRL1 gene contained two exons and one intron. The length of cDNA sequence was 996 bp and the open reading frame(ORF)was 732 bp which encoded a protein of 243 amino acids. Sequence analysis showed that CmCRL1 habored a highly conserved LOB domain,belonging to the LBD/AS2(Lateral Organ Boundaries Domain/Asymmetric Leaves2)family. Multiple alignment analysis revealed that the LOB domain of CmCRL1 had a high conservation with Csa3G396920,Cla005992,AtLBD16,AtLBD29,OsCRL1,ZmRTCS and ZmRTCL,with sequence consistency of 100%,99.02%,74.51%,86.27%,88.24%,89.22% and 80.39%,respectively. Transcriptional expression pattern showed that CmCRL1 was predominantly expressed in roots. The expression level of CmCRL1 in roots was 178.39,7.54,26.95,65.19 and 75.86 times higher than in cotyledons,hypocotyls,epicotyls,leaves and shoot tips. Furthermore,CmCRL1 was identified as being responsive to low temperature,sorbitol,PEG6000 and salt stresses. In addition,CmCRL1 was strongly up-regulated during adventitious root development under waterlogging stress in melon.

Key words: melon, CmCRL1, gene cloning, gene expression

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