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作物杂志,2016, 第4期: 62–67 doi: 10.16035/j.issn.1001-7283.2016.04.010

• 遗传育种·种质资源·生物科技 • 上一篇    下一篇

亚麻糖基转移酶基因LuUGT72E1的克隆与表达分析

袁红梅1,2,郭文栋3,赵丽娟4,于莹2,吴建忠2,程莉莉2,赵东升2,康庆华2,黄文功2,姚玉波2,宋喜霞2,姜卫东2,刘岩2,马廷芬2,吴广文2,关凤芝2   

  1. 1 黑龙江省农业科学院博士后工作站,150086,黑龙江哈尔滨
    2 黑龙江省农业科学院经济作物研究所,150086,黑龙江哈尔滨
    3 黑龙江省科学院自然与生态研究所,150040,黑龙江哈尔滨
    4 黑龙江省农业科学院作物育种所,150086,黑龙江哈尔滨
  • 收稿日期:2016-04-19 修回日期:2016-06-06 出版日期:2016-08-15 发布日期:2018-08-26
  • 通讯作者: 关凤芝
  • 作者简介:作者简介：袁红梅,副研究员,从事作物遗传育种研究
  • 基金资助:
    哈尔滨市科技局创新人才项目(2013RFQYJ162);黑龙江省博士后项目(LBH-Z13181);国家麻类产业技术体系(CARS-19-S03)

Cloning and Expression Analysis of the Glycosyltransferase Gene LuUGT72E1 in Flax

Yuan Hongmei1,2,Guo Wendong3,Zhao Lijuan4,Yu Ying2,Wu Jianzhong2,Cheng Lili2,Zhao Dongsheng2,Kang Qinghua2,Huang Wengong2,Yao Yubo2,Song Xixia2,Jiang Weidong2,Liu Yan2,Ma Tingfen2,Wu Guangwen2,Guan Fengzhi2   

  1. 1 Heilongjiang Academy of Agricultural Sciences Postdoctoral Programme,Harbin 150086,Heilongjiang,China
    2 Industrial Crops Institute,Heilongjiang Academy of Agricultural Sciences,Harbin 150086,Heilongjiang,China
    3 Nature and Ecology Institute,Heilongjiang Academy of Sciences,Harbin 150040,Heilongjiang,China
    4 Crop Breeding Institute,Heilongjiang Academy of Agricultural Sciences,Harbin 150086,Heilongjiang,China
  • Received:2016-04-19 Revised:2016-06-06 Online:2016-08-15 Published:2018-08-26
  • Contact: Fengzhi Guan

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摘要：

以亚麻茎部组织总RNA为模版,通过反转录PCR获得糖基转移酶LuUGT72E1基因的全长开放读码框序列。序列分析结果表明：开放读码框全长1 476bp,共编码491个氨基酸。通过多氨基酸比对发现,该蛋白与亚麻UGT72N2蛋白亲缘关系最近,相似度达99%,与毛果杨、巨桉、木薯、雷蒙德氏棉及拟南芥UGT72E1蛋白的氨基酸序列相似性在58%~51%。分子进化分析表明,该蛋白与拟南芥UGT72E1~3聚为一类,推测该蛋白参与木质素单体的糖基化修饰过程。基因表达谱结果表明,该基因被BR、Brz诱导下调表达。本研究为亚麻的纤维品质改良提供了候选基因。

关键词: 亚麻, 糖基转移酶, 克隆, 表达分析

Abstract:

The total RNA extracted from flax stems was used for RT-PCR, then the full-length open reading frame (ORF) sequence of glycosyltransferase gene (LuUGT72E1) was amplified. The sequence analysis showed that the full-length ORF of LuUGT72E1 consisted of 1 476bp, encoding 491 amino acids. Homology analysis of the deduced amino acid showed 99% identity to UGT72N2 from flax and 58% to 51% identity to UGT72E1 from Populus trichocarpa, Eucalyptus grandis, Manihot esculenta, Gossypium raimondii and Arabidopsis thaliana. The phylogeny analysis showed that this protein clustered with UGT72E1~3 from Arabidopsis and might play key roles in the process of monolignols glucosylation. The gene expression profiling showed that LuUGT72E1 gene could be induced by BR and Brz. This study could provide candidate genes for fibre improvementin flax.

Key words: Flax, Glycosyltransferase, Cloning, Expression analysis

图1

LuUGT72E1基因克隆电泳结果 M：DNA marker(DL2000);1.cDNA模板PCR扩增产物"

图2

LuUGT72E1的核苷酸序列和推导的氨基酸序列"

图3

LuUGT72E1蛋白的预测功能结构域"

图4

糖基转移酶氨基酸序列的多序列比对"

图5

亚麻和拟南芥糖基转移酶基因家族的进化关系"

图6

BR、Brz诱导木质素合成相关基因的表达分析"

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