基于转录组测序的石斛黄酮类化合物生物合成相关基因的克隆与表达分析-植物学专业论文.docxVIP

摘要 摘要 上海师范大学硕士学位论文 大小为43KD左右的蛋白，与预期结果相符合。利用高效液相色谱法对表达的蛋白产物进行 酶活鉴定，结果显示pET32a-DcCHS原核表达产物具有查尔酮合成酶的活性。 3．DcCHS、DcF3H、DcDFR基因的定量表达分析 克隆DcF3H和DcDFR基因的保守序列，并对DcCHS、DcF3H、DcDFR基因进行荧光 定量PCR分析。以普通植株叶为对照，发现三个基因在紫色植株中的表达量高于普通植 株，并且在叶中的表达量高于茎，而在原球茎和组培苗中的表达量较少，这与转录组测序结 果相一致。 关键词：铁皮石斛，黄酮类化合物，转录组测序，基因克隆，荧光定量 万方数据 上海师范大学硕士学位论文 上海师范大学硕士学位论文 Abstract Thesis topic：Cloning and expression analysis of genes involved in Dendrobium flavonoids biosynthesis based on RNA—seq Professional disciplines：Botany Degree applicant：Yi Xiujuan Instruction：Wang Quanxi Abstract Dendrobium eatenatum Lindley is a spiece of Dendrobium SW．，belongs to perennial herbs of the Orchidaceae family．It is of the genuine Dendrobium which collected by”Chinese Pharmacopoeia”．As the secondary metabolism ofD．catenatum，flavonoids is the primary bioactive component which has the effect of antioxidation，anti—aging and anti virus and so on，and it also as the main component which can resist diseases and stresses forthe plant itself．Currently the study of flavonoids in D，catenatum mainly concentrated component identification and species analysis． However，the details of flavonoids biosynthesis have not been effectively explored．This study respectively from three aspects to provide basis for the study of flavonoids metabolic pathways in Dendrobium SW．．The main results ofthe study include the following three aspects： 1．The RNA-seq of Dendrobium catenatum RNA—seq analysis of the leaves of common and purple plants in artificial cultivation．In this study，we analyzed RNA isolated from D．catenatum using Hiseq 2000 Illumina platform to generate 64，1 95 unigenes，the total length，average length，N50，and GC content of urfigenes are 66，479，063 bp，1,035 bp，1,742 bp，and 42．47％，respectively．A total of35．92％ofthe unigenes were annotated with Kyoto Encyclopedia of Genes and Genomes(K．EGG)terms，and a detailed view of flavonoids biosynthesis was obtained．Differential expression analysis results under the control of common leaves showed that t