四川农业大学学报 ›› 2015, Vol. 33 ›› Issue (01): 6-12.doi: 10.16036/j.issn.1000-2650.2015.01.002

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菊花CmLhcb1基因的克隆及其转基因植株的筛选

韩霜1, 逯昀2   

  1. 1. 商丘师范学院生命科学学院, 植物与微生物互作重点实验室, 河南 商丘 476000;
    2. 商丘职业技术学院成教学院, 河南 商丘 476000
  • 收稿日期:2014-10-21 出版日期:2015-03-31 发布日期:2017-02-28
  • 作者简介:韩霜,博士,讲师,主要从事植物光合作用方面研究,E-mail:htshd-012@163.com。
  • 基金资助:
    河南省科技厅基础与前沿项目(142300410321);河南省重点科技攻关项目(142102110184)

Cloning of the CmLhcb1 Gene and Screening of Transgenic Plant of Chrysanthemum morifolium

HAN Shuang1, LU Yun2   

  1. 1. Key Laboratory of Plant-Microbe interactions, Department of Life Science, Shangqiu Normal University, Shangqiu 476000, Henan, China;
    2. College of Adult education, Shangqiu Polytechnic, Shangqiu 476000, Henan, China
  • Received:2014-10-21 Online:2015-03-31 Published:2017-02-28

摘要: [目的] 克隆CmLhcb1基因并转到拟南芥中,从而探索此基因的功能。[方法] 以菊花品种'清露'幼苗为试验材料,采用RT-PCR和RACE技术克隆获得其叶绿素a/b结合蛋白基因CmLhcb1,并进一步构建表达载pEarleyGate103-CmLhcb1,经农杆菌介导法,筛选获得3个T3代阳性转基因拟南芥植株。[结果] 该基因序列编码的蛋白与其他物种相似度高,主要差异存在于C端前50个氨基酸。基因表达特征分析表明,该基因在15%光照和赤霉素条件下,表达量明显增加(P<0.05),叶片中表达量最大,根中表达量最低,白天的表达量比晚上的高30倍。[结论] 菊花CmLhcb1基因受光和赤霉素诱导,有组织表达特异性。

关键词: 菊花, 基因, 克隆, 表达载体

Abstract: [Objective] The gene of CmLhcb1 was cloned and transferred into Arabidopsis. The function of the gene was explored. [Method] Chrysanthemum named ‘Puma sunny’ seeding was used as experimental material. We use PCR (Polymerase Chain Reaction)method in vitro to amplify chlorophyll a/b binding protein gene (CmLhcb1). The plant expression vector of chrysanthemum pEarleyGate103-CmLhcb1 was constructed. We got 3 T3 positive transgenic Arabidopsis plants by agrobacterium mediated method. [Results] There was high similarity between the protein coding sequence of the gene and the those of other species. The major differences were in the former 50 amino acids of C terminal. The analysis of gene expression profile showed that the gene expression increased significantly under the conditions of 15% light and gibberellin (P<0.05). The expression was the greatest in leaves but was the lowest in root. The expression of day was 30 times higher than that of the evening. [Conclusion] The CmLhcb1 gene of C. morifolium had significant specific tissue expression when it was induced by light or gibberellin.

Key words: Chrysanthemum morifolium, gene, cloning, expression vector

中图分类号: 

  • S682.1+1