水母雪莲花青素合成相关基因的克隆及表达  

Cloning and Expression of Anthocyanin-related Gene in Saussurea medusa Maxim.

作　　者：郭佳磊 王联星 李凤珍 史国民 赵伟民 何涛 GUO Jialei;WANG Lianxing;LI Fengzhen;SHI Guomin;ZHAO Weimin;HE Tao(School of Ecol-Environmental Engineering, Qinghai University, Xining 810016, China;2Key Laboratory of Landscape Plants of Qinghai Province, Xining 810016, China;3State Key Laboratory of Plateau Ecology and Agriculture, Xining 810016, China)

机构地区：[1]青海大学生态环境工程学院,西宁810016 [2]青海省园林植物与观赏园艺重点实验室,西宁810016 [3]省部共建三江源生态与高原农牧业国家重点实验室,西宁810016

出　　处：《西北植物学报》2020年第11期1840-1846,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31960222,31360095)。

摘　　要：为研究高山植物水母雪莲对强紫外辐射的分子适应机制,采用RT-PCR结合RACE技术从水母雪莲中克隆了参与调控花青素合成的相关基因(SmMYB1),并采用hi-TAIL PCR方法扩增了该基因的启动子序列。结果表明:(1)序列分析显示,SmMYB1基因cDNA序列(GenBank登录号MT188353)全长1011 bp,编码269个氨基酸,gDNA序列含有2个内含子和3个外显子。(2)生物信息学分析显示,SmMYB1基因编码蛋白和菊科MYB蛋白亲缘关系最近,含有保守的[DE]Lx(2)[RK]x(3)Lx(6)Lx(3)R和ANDV两个模体,属于拟南芥MYB第六亚族。SmMYB1基因启动子(GenBank登录号MT188354)全长1407 bp,含有多个光响应元件。(3)荧光定量分析发现,SmMYB1基因在根、茎、叶和花中均表达,且在花中的表达量最高;在紫外胁迫下,SmMYB1基因的表达量在4 h达到最高,随后逐渐降低。研究推测SmMYB1基因可能参与调控水母雪莲花青素的合成以及对紫外辐射的响应过程。In order to study the molecular adaptation mechanism of the alpine plant Saussia medusa to high ultraviolet radiation,we cloned the cDNA and promoter sequences of anthocyanin-related SmMYB1 gene by RT-PCR combined with RACE methods and hi-TAIL PCR.The results show that:(1)the length of SmMYB1 cDNA(GenBank:MT188353)is 1011 bp encoding 269 amino acids,and the sequence of gDNA contains two introns and three extrons.(2)Bioinformatics analysis show SmMYB1 protein has more high similarity with MYB in Asteraceae contained the[DE]Lx(2)[RK]x(3)Lx(6)Lx(3)R and ANDV motifs,and belongs to the sixth subfamily of Arabidopsis thaliana.The length of SmMYB1 promoter(GenBank:MT188354)is 1407 bp,which contains many light responsive elements.(3)qRT-PCR shows that SmMYB1 gene is expressed in roots,stems,leaves and flowers,and the highest expression emerges in the flowers.Under ultraviolet radiation stress,the expression of SmMYB1 reaches the highest at 4 h,and then decreases gradually.It is speculated that SmMYB1 can regulate anthocyanin synthesis and is involved in the UV responsive pathway in S.medusa.

关 键 词：水母雪莲 SmMYB1基因 花青素 启动子 表达分析 

分 类 号：Q785[生物学—分子生物学] Q786