决明不同组织莽草酸激酶基因的表达模式与蒽醌含量分析  

Tissue-specific expression pattern and total anthraquione contents analysis of shikimate kinase from Cassia obtusifolia

作　　者：李洋洋[1] 李卓彬[1] 俞继华[1] 李娟娟[1] 王万军[1] 周嘉裕[1] 廖海[1] 

机构地区：[1]西南交通大学生命科学与工程学院,成都610031

出　　处：《生物学杂志》2016年第3期30-33,共4页Journal of Biology

基　　金：国家自然科学基金项目(NAC特异基因调控铁皮石斛原球茎原分生组织形成的分子机理研究;No.31271302);西南交通大学大学生科研训练项目(2014065)

摘　　要：分析莽草酸激酶基因在决明(Cassia obtusifolia L.)不同组织中的表达模式,同时测定不同组织中总蒽醌的含量,为研究莽草酸激酶在决明蒽醌生物合成中的作用提供理论依据。提取决明种子、愈伤组织与成熟叶等10个不同组织的总RNA,反转录合成c DNA,采用荧光定量PCR检测莽草酸激酶基因在不同组织的表达情况。同时,采用超声波强化法提取决明不同组织的总蒽醌,通过分光光度法测定其含量。莽草酸激酶基因在决明的根、愈伤组织与子叶中的表达量较高,在种子、果荚与茎中表达量较低。决明的种子、果荚、花及根中蒽醌类化合物含量较高,而成熟叶和胚轴则不含。决明中蒽醌的生物合成涉及多个组织,其生物合成的早期阶段(包括莽草酸激酶催化的反应)主要发生在根与子叶等组织,形成的中间产物运输到种子等组织中,完成生物合成的后期阶段并积累蒽醌终产物,为合理利用决明不同药用部位提供理论依据。Tissue-specific expression pattern of shikimate kinase from Cassia obtusifolia L. was analyzed and then the total anthraquione content in different organs was determined,which aimed at providing a theoretical basis for the study of the role of shikimate kinase from Cassia obtusifolia in anthraquinone biosynthesis. The total RNA of nonage seed,callus and mature leaves and so on of C. obtusifolia were extracted and subjected for c DNA synthesis. The expression quantitative analysis of Co SK in different tissues was estimated by real-time PCR. At the same time the anthraquione was extracted by ultrasonic extracting method,and the total anthraquione contents were determined by UV. Shikimate kinase showed a significantly high expression level in the root,callus and cotyledon of C. obtusifolia and a relatively low expression level in stem,pod and nonage seed. Anthraquione illustrated high content in seeds,pod,flowers and roots of C. obtusifolia,whereas mature leaves and hypocotyls did not find anthraquinone. The biosynthesis of C. obtusifolia in anthraquinone involved multiple organizations and in early stages（ including shikimate kinase-catalyzed reaction） occured mainly in roots and mature leaves and other tissues. The intermediate products were transported to seeds and other tissues and completed the later stages of the biosynthesis and accumulation of anthraquinone final product,which was supposed to provide a guideline for the rational utilization of different medicinal parts of C. obtusifolia.

关 键 词：决明 莽草酸激酶 组织差异表达 总蒽醌 含量测定 

分 类 号：Q344.13[生物学—遗传学]