绿色杜氏藻不同β.pdf

Hereditas (Beijing) 2017 年 2 月, 39(2): 156 ―165 研究报告 绿色杜氏藻不同β-胡萝卜素羟化酶基因家族胁迫 应答模式研究 朱帅旗，龚一富，章丽，俞凯，王何瑜，严小军 宁波大学海洋学院，宁波 315211 摘要 : β-胡萝卜素羟化酶(β-carotenoid hydroxylase, CHYB)是植物类胡萝卜素生物合成途径中的一个重要限速 酶。本研究对绿色杜氏藻转录组测序数据进行分析，获得2 条 β-胡萝卜素羟化酶家族基因chyb1 和chyb2 。采 用染色体步移法分别克隆并获得了绿色杜氏藻chyb1 和chyb2 基因的启动子序列，全长分别为 1080 bp(GenBank 登录号：KY012338)和 1155 bp (GenBank 登录号：KY012339) 。利用Plantcare 软件分析两个启动子的顺式作用 元件，结果表明绿色杜氏藻chyb1 基因启动子含有与甲基茉莉酸、花生四烯酸、水杨酸等非生物胁迫相关的顺 式作用元件，而绿色杜氏藻chyb2 基因启动子含有与光照相关的顺式作用元件。通过 qRT-PCR 分析了绿色杜 氏藻CHYB 基因家族在不同胁迫下的基因表达水平，结果表明该基因家族的基因表达水平与启动子调控相关， 且不同的家族基因应答不同的胁迫。 关键词 : 绿色杜氏藻；β-胡萝卜素羟化酶；基因家族；启动子；胁迫 Expression analysis of different β-carotenoid hydroxylase gene families in stress response in Dunaliella viridis Shuaiqi Zhu, Yifu Gong, Li Zhang, Kai Yu, Heyu Wang, Xiaojun Yan School of Marine Sciences, Ningbo University, Ningbo 315211, China Abstract: β-carotenoid hydroxylase (CHYB) is an important rate-limiting enzyme in the biosynthesis of plant ca- rotenoid. In this study, chyb1 and chyb2, two gene families in Dunaliella viridis were obtained by RNA-seq. The fragment of promoters of CHYB family genes, 1 080 bp for chyb1 (GenBank No. KY012338) and 1 155 bp for chyb2 (GenBank No. KY012339) were cloned by the Genome Walking Technology, respectively. Cis-acting elements of two promoters were analyzed by Plantcare soft. The results show that the chyb1 gene promoter contains more cis-acting elements in responses to abiotic stresses, such as methyl jasmonate, arachidonic acid, acetylsalicylic acid, and so on. On the other hand, the chyb2 promoter contain s more cis-acting elements in response to light stress. qRT-PCR results 收稿日期: 2016−09−22 ; 修回日期: 2016−12−04 基金项目: 浙江省科技厅重点科技创新团队项目(编号：2012R10029-07, 2010R50029) ，宁波科技攻关项目(编号：2013C10018)和宁波大学优秀学位论 文培育基金资助项目(编号：py2014020) 资助[Supported by Key Scientific an