罗汉果乙烯合成酶SgA CS1基因的克隆与表达分析

发布时间：2018-09-06 18:59

【摘要】：在转录组unigene序列基础上,结合RACE技术,从罗汉果总RNA中克隆乙烯合成关键酶-1-氨基环丙烷-1-羧酸合成酶1(ACS1)全长cDNA,同时,应用hi TAIL PCR技术获得Sg ACS1的DNA全长。通过实时荧光定量PCR检测该基因在雄株、雌株以及Ag~+处理后的雌株在取/授粉前后的叶、茎、芽、花蕾、花中的相对表达量。克隆得到1 749 bp的cDNA全长,最长开放阅读框为1 530 bp,编码509个氨基酸,命名为Sg ACS1(GenBank登录号为KX620760),编码蛋白与同源物种苦瓜的相应蛋白序列相似性为86%,该蛋白具有磷酸吡哆醛转移酶结构域和转氨酶结合结构域,属于天冬氨酸转氨酶家族;获得Sg ACS1的DNA全长为3 245 bp,其中含有3个内含子,该内含子和5'UTR含有多个高水平转录调控因子、激素响应元件和环境胁迫相关的作用元件,暗示该基因参与转氨反应、生物体内乙烯的合成代谢等生物学过程,并且与其他激素共同发挥作用维持机体的正常生命活动。实时荧光定量PCR结果表明,Sg ACS1在罗汉果各个部位和时期均有表达,其中在授粉后的雌株花蕾中相对表达量最高,其次是授粉前后雌株的花和取粉前后雄株的芽,而Ag~+处理后的雌株在授粉后的花蕾和芽中的相对表达量最低,说明该暗示罗汉果Sg ACS1参与花芽形成、雄蕊原基分化、蕾中雄蕊发育以及雌花形成,Sg ACS1在雌株蕾中受到授粉或银离子诱导上调表达。
[Abstract]:On the basis of transcriptome UniGene sequence and RACE technique, the full-length cDNA of the key enzyme of ethylene synthesis - 1-aminocyclopropane-1-carboxylic acid synthase 1 (ACS1) was cloned from grosvenorii total RNA, and the full-length of Sg ACS1 was obtained by hi-TAIL PCR. 1 749 BP cDNA was cloned from leaves, stems, buds, buds and flowers before and after pollination. The longest open reading frame was 1 530 BP and encoded 509 amino acids. It was named Sg ACS1 (GenBank login number KX620760), and the sequence similarity of the encoded protein was 86% with that of the homologous bitter gourd. Transferase domain and transaminase binding domain belong to aspartate aminotransferase family; the total length of DNA obtained from Sg ACS1 is 3 245 bp, which contains three introns. The intron and 5'UTR contain many high-level transcription regulators, hormone response elements and environmental stress-related elements, suggesting that the gene is involved in transamination reaction, organisms. Real-time fluorescence quantitative PCR showed that Sg ACS1 was expressed in all parts and stages of grosvenoria grosvenorii, and the relative expression of Sg ACS1 was the highest in the female buds after pollination, followed by the flowers of the female plants before and after pollination. The relative expression of Sg ACS1 in the buds and buds of male plants before and after pollination was the lowest, indicating that Sg ACS1 was involved in flower bud formation, stamen primordium differentiation, stamen development and female flower formation. Sg ACS1 was up-regulated by pollination or silver ion in female buds.
【作者单位】：广西大学农学院;广西药用植物园广西药用资源保护与遗传改良重点实验室;九江市第三人民医院;
【基金】：国家自然科学基金项目(31260359) 广西大学博士启动项目(XBZ110586)共同资助
【分类号】：S668.9;Q943.2

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