中国农业科学 ›› 2017, Vol. 50 ›› Issue (17): 3274-3285.doi: 10.3864/j.issn.0578-1752.2017.17.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

拟南芥仅含START结构域亚家族基因特性与功能分析

闫青地,赵亚林,张昊,高静,高梦烛,王倩,王蕊,王凤茹,董金皋   

  1. 河北农业大学生命科学学院/河北省植物生理和分子病理学重点实验室,河北保定071001
  • 收稿日期:2017-03-03 出版日期:2017-09-01 发布日期:2017-09-01
  • 通讯作者: 王凤茹,E-mail:wfr15931945160@126.com。董金皋,E-mail:dongjingao@126.com
  • 作者简介:闫青地,E-mail:1073886780@qq.com;赵亚林,E-mail:1520469399@qq.com。闫青地和赵亚林为同等贡献作者。
  • 基金资助:
    河北省自然科学基金(C2017204060)

Analysis on Gene Feature and Function of the Subfamily Members Containing START Domain Only in Arabidopsis thaliana

YAN QingDi, ZHAO YaLin, ZHANG Hao, GAO Jing, GAO MengZhu, WANG Qian, WANG Rui, WANG FengRu, DONG JinGao   

  1. College of Life Science, Agricultural University of Hebei/Key Laboratory of Hebei Province for Plant Physiology and Molecular Plant Pathology, Baoding 071001, Hebei
  • Received:2017-03-03 Online:2017-09-01 Published:2017-09-01

摘要: 【目的】分析拟南芥中仅含START结构域(the lipid/sterol-binding StAR-related lipid transfer protein domains)亚家族成员的启动子元件及表达特性,阐明启动子元件与表达特性的相互关系,为明确仅含START结构域亚家族的生物学功能,解析植物生长发育机理、更好地促控植物生长提供理论依据。【方法】利用生物信息学方法分析仅含START结构域亚家族6个成员的亲缘关系及各自启动子区所包含的所有元件,对启动子元件的功能进行分析和分类;利用real-time PCR方法研究亚家族成员的表达特性;用突变体分析确认基因的功能;分析通过启动子元件分析预测的基因功能与通过突变体分析确认的基因功能间的相关性,为基因的功能和作用机制分析提供科研思路和方法。【结果】生物信息学分析表明,拟南芥中仅含START结构域亚家族成员共6个,分为3个分支,每个分支一对基因,每一对基因中均有一个基因的启动子中含有高转录水平元件5′ UTR Py-rich stretch,6个亚家族成员的启动子区均含有多个光、激素和逆境响应元件,也存在各自特异的响应元件;用real-time PCR分析基因的表达量,结果与生物信息学预测结果一致:At3g13062的表达量明显高于同分支的At1g55960,At3g23080的表达量明显高于同分支的At4g14500,At1g64720的表达量明显高于同分支的At5g54170。组织定位分析表明亚家族成员有特异的时空表达特性:At3g13062和At1g55960在幼苗期就有明显表达,但At3g13062主要在子叶、下胚轴和根中表达,而At1g55960主要在根中表达量较多;在成苗期拟南芥中,At3g13062主要在叶片中表达,在根中表达量较少;而At1g55960在成苗期拟南芥的叶片和根系的表达量均较多;At3g23080和At4g14500在幼苗期表达量较少,而在成苗期表达量增加,At3g23080主要分布于莲座叶中,At4g14500主要分布于莲座叶叶柄部。上述结果表明仅含START结构域亚家族成员可能在拟南芥不同时期和不同的部位分别行使着功能。利用生物信息学对启动子元件分析发现,各个基因均具有胚乳特意表达的Skn-1 motif元件,暗示着该家族基因可能参与调控胚乳的发育,进而影响种子的活力和萌发;通过对At3g23080和At4g14500的T-DNA插入突变体种子的分析表明,At3g23080和At4g14500表达量下降均会导致种子活力和萌发率的下降,这与生物信息学预测结果一致。【结论】拟南芥中仅含START结构域的亚家族有6个成员;6个亚家族成员可能在光、激素和逆境调控的发育过程中具有重要的作用;含START结构域的亚家族6个成员虽然同源关系较近,但具有各自的时空表达特性,在拟南芥的不同组织和不同发育时期执行着各自的功能。基因启动子元件与基因功能有直接关系,启动子区具有胚乳特意表达Skn-1 motif元件的At3g23080和At4g14500表达量下降均会导致种子活力和萌发率下降。

关键词: 拟南芥, START结构域, 启动子元件, 表达特性, 基因功能

Abstract: 【Objective】 The objective of this study is to analyze the promoter elements and expression feature of the genes coding the subfamily containing the START domain (the lipid/sterol-binding StAR-related lipid transfer protein domains) only in Arabidopsis thaliana, expound the relationship between the cis-acting element and the gene function, and the results of study will provide an important theoretical basis for clarifying the biological function of the genes, analyzing the plant growth and development mechanism and promoting or suppressing plant growth better. 【Method】 Using bioinformatics method to analyze the genetic relationship of 6 members and all the elements contained in their respective promoter regions. The function of the promoter elements of subfamily were analyzed and classified, and the expression features and the regulation factors of these subfamily members were predicted. Real-time PCR method was employed to determine the expression characteristics of subfamily members, clarify the relationship between the test results and the conclusions of promoter analysis, thus providing scientific research ideas and direction for analysis of the function and mechanism of genes. The relationship between the gene promoter element and function was confirmed after analysis of the gene mutation type surface. 【Result】 There are 6 members belong to the subfamily containing START domain only in A. thaliana, divided into three branches, every branch has a pair of genes, each pair of genes has a promoter containing high transcription element 5′ UTR Py-rich stretch. The promoter regions of the 6 subfamily members contain multiple common response elements, such as light, hormone and stress, and also have specific response elements for their own. The real-time PCR results of the genes expression level showed that the expression level of At3g13062 was significantly higher than the same branch gene At1g55960, At3g23080 was significantly higher than the same branch gene At4g14500, At1g64720 was significantly higher than the same branch gene At5g54170. All the subfamily members have higher expression levels at seedling stage, but less expression levels at ageing stage, and have its own specific expression of space and time characteristics. In the gene promoter analysis of the family containing START domain structure only, it was found that each gene promoter has a cis-acting element Skn-1 motif which was specially expressed in endosperm, implied that the family genes may be involved in regulation of endosperm development. Seeds analysis of At3g23080 and At4g14500 T-DNA insertion mutant showed that At3g23080 and At4g14500 expression decreased and caused a decline in the seed vigor. Conclusion There are 6 members in the subfamily containing START domain only. Six members of the family may be play an important role in the response process to light, hormones and stress. the 6 family members have a close kinship, but their expression characteristics of time and space are different and carry out their respective functions in different development periods of A. thaliana. There is a direct relationship between gene promoter elements and the gene function, and the promoter region with endosperm expressed specially element Skn1 motif in At3g23080 and At4g14500, so the decreased expression of At3g23080 or At4g14500 caused a decline in seed vigor.

Key words: Arabidopsis thaliana, START domain, promoter elements; expression feature, gene function