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热带作物学报 ›› 2020, Vol. 41 ›› Issue (5): 964-970.doi: 10.3969/j.issn.1000-2561.2020.05.016

• 生物技术与组织培养 • 上一篇    下一篇

龙眼DlICE1基因克隆及低温胁迫表达分析

霍雯,李佳蜜,徐小萍,赖瑞联,陈晓慧,林玉玲,陈裕坤,赖钟雄()   

  1. 福建农林大学园艺植物生物工程研究所,福建福州 350002
  • 收稿日期:2019-04-28 修回日期:2019-09-09 出版日期:2020-05-25 发布日期:2020-06-15
  • 通讯作者: 赖钟雄 E-mail:laizx01@163.com
  • 作者简介:霍 雯(1993—),女,硕士研究生,研究方向:果树生物技术。
  • 基金资助:
    国家自然科学基金(31572088);福建省高原学科建设项目(102/71201801101);福建农林大学科技创新基金

Cloning and Expression of Low Temperature Stress of DlICE1 in Dimocarpus longan Lour.

HUO Wen,LI Jiami,XU Xiaoping,LAI Ruilian,CHEN Xiaohui,LIN Yuling,CHEN Yukun,LAI Zhongxiong()   

  1. Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou, Fujian 35002, China
  • Received:2019-04-28 Revised:2019-09-09 Online:2020-05-25 Published:2020-06-15
  • Contact: LAI Zhongxiong E-mail:laizx01@163.com

摘要:

本研究以龙眼胚性愈伤组织为材料,根据龙眼转录组数据库,采用RACE-PCR和RT-PCR技术获取龙眼DlICE1 cDNA全长序列,并对其进行生物信息学分析与低温胁迫下表达分析。结果表明,龙眼DlICE1 cDNA全长为2327 bp,其中开放式阅读框(ORF)序列长1614 bp,共编码537个氨基酸。生物信息学分析表明,龙眼DlICE1编码的蛋白质属于不稳定的弱酸性蛋白,不具有典型的信号肽,且不属于分泌蛋白;亚细胞定位表明其位于细胞核内,与甜橙的同源蛋白亲缘关系最近;实时荧光定量PCR表明,龙眼DlICE1能够有效响应低温胁迫,低温诱导其表达量提高。研究结果显示,龙眼DlICE1基因参与龙眼胚性愈伤组织抗寒过程。

关键词: 龙眼, 胚性愈伤组织, DlICE1基因, 基因克隆, 实时荧光定量PCR

Abstract:

In this experiment, the embryogenic callus of longan were used as the materials, the cDNA sequence of DlICE1 was cloned from longan embryogenic callus by the method of RACE-PCR and RT-PCR, then it was analyzed by bioinformatical methods and expression analysis under low temperature stress. The full-length sequence of DlICE1 was 2327 bp, which contained an 1614 bp open reading frame encoded 537 amino acids. The bioinformatics analysis indicated that DlICE1 belonged to the unstable acidic protein, which didn’t has a typical signal peptide and wasn’t a secreted protein; subcellular localization indicated that it is located in the nucleus, which was closest to the homologous protein of sweet orange. The qRT-PCR results indicated that the longan DlICE1 could effectively respond to low temperature stress and induce increased expression at low temperature. In summary, the study suggests that the longan DlICE1 gene may be involved in the cold resistance in longan embryogenic callus.

Key words: Dimocarpus longan Lour., embryogenic callus, DlICE1 gene, gene cloning, real-time fluorescent quantitative PCR

中图分类号: 

  • S667.2