蒙古黄芪叶绿体基因组研究

发布时间：2018-05-14 21:12

本文选题：蒙古黄芪 + 叶绿体基因组　；参考：《山西农业大学》2016年博士论文

【摘要】：在亚洲,植物黄芪(Astragalus membranaceus)是一种重要的药用植物。用于中药生产加工的主要是黄芪的两个变种,蒙古黄芪(A. membranaceus (Fisch.) Bunge var. mongholicus (Bunge) P.K. Hsiao)和膜荚黄芪(A. membranaceus (Fisch.) Bunge var. membranaceus)。黄芪的一些其他变种也经常被用于中药生产。因此为了规范生产和科学研究,急需开发可用于分辨不同黄芪变种的高分辨率的遗传标记。本研究中,使用第二代高通量测序技术对蒙古黄芪叶绿体基因组进行了测序和分析。测序平台为hiseq2000,黄芪叶绿体基因组短读序拼接使用Abyss软件,叶绿体基因组基因注释使用CPGAVAS软件,重复序列分析使用MISA软件、Tandem Repeats软件和REPuter软件。之后对组装注释后完整的黄芪叶绿体基因组进行系统进化分析和比较基因组学分析。组装后得到的完整蒙古黄芪叶绿体基因组全长为123,582bp。蒙古黄芪叶绿体基因组结构并不呈现典型的四段式结构,而是只具有单个反向重复拷贝(IR区)。蒙古黄芪叶绿体基因组编码110个基因,包括76个蛋白编码基因、30个tRNA基因和4个rRNA基因。在使用PCR技术填补经组装得到的大片段之间缺口区的过程中,发现并验证了蒙古黄芪叶绿体基因组上5个种内的高可变位点,其中3个还表现出胞质异质性。进一步分析发现,蒙古黄芪叶绿体基因组于典型的被子植物叶绿体基因组相比较存在三个基因缺失的现象和两个大片段的倒位现象。对已公开完整叶绿体基因组序列的36种豆科蝶形花亚科植物的叶绿体基因组序列进行比较基因组学分析。结果显示,蝶形花亚科植物的叶绿体基因组在种间和种内水平具有动态变化的特点,这些变化包括大量的高可变位点、频繁的基因缺失和大片段序列的倒位现象。本研究对中药材黄芪重要的来源植物蒙古黄芪经行完整叶绿体基因组的测序组装并进行了分析。其研究结果为基于蒙古黄芪叶绿体基因组序列开发高分辨率分子标记奠定了基础,以满足黄芪的系统进化和群体遗传学研究需要。同时也有助于研究和了解被子植物,尤其是豆科蝶形花亚科植物叶绿体基因组复杂的进化历史。
[Abstract]:Astragalus membranaceus is an important medicinal plant in Asia. Two varieties of Radix Astragali, Astragalus mongolicus A. membranaceus Fisch., were mainly used in the production and processing of traditional Chinese medicine. Bunge var. mongholicus BungeP.K. Hsiao and Astragalus membranaceus A. membranaceus Fisch. Bunge var. membranaceus. Other varieties of Astragalus are also often used in the production of traditional Chinese medicine. Therefore, in order to standardize production and scientific research, it is urgent to develop high resolution genetic markers that can be used to distinguish different varieties of Astragalus membranaceus. In this study, the chloroplast genome of Astragalus mongolicus was sequenced and analyzed by the second generation high-throughput sequencing technique. The sequencing platform was hiseq2000. Abyss software was used for short reading sequence splicing of chloroplast genome of Astragalus membranaceus, CPGAVAS software was used for chloroplast genome gene annotation, and MISA software Tandem Repeats software and REPuter software were used for repeated sequence analysis. The complete chloroplast genome of Astragalus membranaceus was analyzed by phylogenetic analysis and comparative genomics analysis. The complete chloroplast genome of Astragalus mongolicus was 123582 BP. The chloroplast genome of Astragalus mongolicus does not exhibit a typical four-segment structure, but only has a single reverse repeat copy of the IR region of Astragalus mongolicus. The chloroplast genome of Astragalus mongolicus encodes 110 genes, including 76 protein coding genes, 30 tRNA genes and 4 rRNA genes. In the process of filling the gap region between large fragments by PCR technique, five high variable loci in chloroplast genome of Astragalus mongolicus were found and verified, and three of them showed cytoplasmic heterogeneity. Further analysis showed that the chloroplast genome of Astragalus mongolicus had three gene deletions and two large fragments in comparison with the chloroplast genome of typical angiosperms. The chloroplast genomes of 36 species of sphenoid subfamily in legume were analyzed by comparative genomics. The results showed that the chloroplast genomes of the subfamily Butterfly showed dynamic changes at interspecific and intra-specific levels, including a large number of high variable loci, frequent gene deletions and large fragment sequence inversion. In this study, the complete chloroplast genome of Astragalus mongolicus (Astragalus mongolicus), an important source of Astragalus membranaceus, was sequenced and analyzed. The results laid a foundation for the development of high-resolution molecular markers based on the chloroplast genomic sequence of Astragalus mongolicus, in order to meet the needs of systematic evolution and population genetics of Astragalus membranaceus. It is also helpful to study and understand the complex evolutionary history of chloroplast genome in angiosperms, especially in the sphenoid subfamily of legumes.
【学位授予单位】：山西农业大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：S567.239

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