三角褐指藻crtiso基因克隆与表达调控研究  

Cloning and Expression Analysis of Carotenoid Isomerase Gene From Phaeodactylum tricornutum

作　　者：管悦琳 龚一富[1] 朱帅旗 俞凯[1] 王何瑜[1] 严小军[1] GUAN Yuelin;GONG Yifu;ZHU Shuaiqi;YU Kai;WANG Heyu;YAN Xiaojun(Key Laboratory of Applied Marine Biotechnology,Ministry of Education/School of Marine Sciences,Ningbo University,Ningbo,Zhejiang 315211)

机构地区：[1]宁波大学海洋学院/教育部应用海洋生物技术重点实验室,浙江宁波315211

出　　处：《核农学报》2018年第11期2098-2106,共9页Journal of Nuclear Agricultural Sciences

基　　金：国家科技部星火计划项目(2015GA701001;2014GA701020);浙江省科技厅重点科技创新团队项目(2012R10029-07;2010R50029);宁波科技攻关项目(2013C10018)

摘　　要：为探索三角褐指藻岩藻黄素的生物合成与crtiso基因表达调控的关系,本研究通过转录组测序获得了三角褐指藻crtiso基因cDNA全长序列,并研究了乙酰水杨酸(ASA)、花生四烯酸(AA)、甲基茉莉酸(MeJA)和硫酸铈铵(ACS)4种诱导子不同浓度处理对三角褐指藻crtiso基因表达的影响。结果表明,三角褐指藻crtiso cDNA全长2 116 bp,开放阅读框(ORF)1 902 bp,编码635个氨基酸。crtiso蛋白为亲水性不稳定蛋白,相对分子质量67 803.00 Mr,理论等电点7.14,蛋白质二级结构中的主要构成元件是α螺旋、β折叠和无规则卷曲,并存在保守区域和结构域。系统进化树分析表明,三角褐指藻crtiso蛋白与海虹束毛藻亲缘关系较近。诱导子表达结果表明,ASA、AA、MeJA和ACS均能显著提高三角褐指藻crtiso基因的表达水平,当ASA、AA、MeJA和ACS质量浓度分别为10 mg·L^(-1)、0.1 mg·L^(-1)、50μmol·L^(-1)和0.2 mg·L^(-1)时,三角褐指藻crtiso基因表达量最高。相关性分析表明,三角褐指藻crtiso基因表达量与岩藻黄素含量呈线性关系,表明三角褐指藻岩藻黄素的生物合成是通过调控crtiso基因的表达来实现的,本试验结果为进一步研究岩藻黄素的合成代谢提供了重要的依据。In order to explore the relationship between the biosynthesis of fucoxanthin and the regulation of crtiso gene expression,the full-length cDNA sequence of crtiso was obtained by transcriptome sequencing and the effects of acetylsalicylic acid（ ASA）,arachidonic acid（ AA）,methyl jasmonate（ MeJA） and ammonium ceric sulfate（ ACS） on the crtiso gene expression of Phaeodactylum tricornutum at different concentrations were studied. The results showed that the crtiso cDNA was 2 116 bp in length and 1 902 bp in open reading frame（ ORF）,encoding 635 amino acids. crtiso protein is a hydrophilic labile protein,relative molecular mass is 67 803. 00 Mr,with the theoretical isoelectric point of7. 14,the main component of protein secondary structure is α-helix,β-sheet and random curl,with conserved regions and domains. The phylogentic tree analysis indicated that the crtiso protein of Phaeodactylum tricornutum had a close genetic relationship with Cyanbacteria. The results of elicitor expression indicated that ASA,AA,MeJA and ACS could significantly increase the expression level of crtiso gene. When the concentration of ASA,AA,MeJA and ACS were 10 mg·L^-1,0. 1 mg·L^-1,50 μmol·L^-1 and 0. 2 mg·L^-1 respectively,the expression of crtiso gene was the highest.Correlation analysis showed that the expression level of crtiso gene and the content of fucoxanthin was linear related,indicating that the biosynthesis of fucoxanthin triptolide by regulating crtiso gene expression. This study provides an important basis for further studing the synthesis of fucoxanthin in the future.

关 键 词：三角褐指藻 类胡萝卜素异构酶 岩藻黄素 基因克隆 表达分析 

分 类 号：Q943.2[生物学—植物学]