Park Chang Ha, Yeo Hyeon Ji, Park Ye Eun, Baek Seung-A, Kim Jae Kwang, Park Sang Un
Department of Crop Science, Chungnam National University, 99 Daehak-ro, Yuseong-gu, Daejeon 305-754, Korea.
Division of Life Sciences and Convergence Research Center for Insect Vectors, Incheon National University, Incheon 22012, Korea.
Biology (Basel). 2019 Aug 29;8(3):63. doi: 10.3390/biology8030063.
belongs to the Amaryllidaceae family and is a bulbous plant native to South Korea, China, and Japan. Galantamine, a representative alkaloid of Amaryllidaceae plants, including . , exhibits selective and dominant acetylcholinesterase inhibition. In spite of the economic and officinal importance of . , the molecular biological and biochemical information on . is relatively deficient. Therefore, this study provides functional information of . , describe galantamine biosynthesis in the various organs, and provide transcriptomic and metabolic datasets to support elucidation of galantamine biosynthesis pathway in future studies. The results of studies conducted in duplicate revealed the presence of a total of 325,609 and 404,019 unigenes, acquired from 9,913,869,968 and 10,162,653,038 raw reads, respectively, after trimming the raw reads using CutAdapt, assembly using Trinity package, and clustering using CD-Hit-EST. All of the assembled unigenes were aligned to the public databases, including National Center for Biotechnology Information (NCBI) non-redundant protein (NR) and nucleotide (Nt) database, SWISS-PROT (UniProt) protein sequence data bank, The Arabidopsis Information Resource (TAIR), the Swiss-Prot protein database, Gene Ontology (GO), and Clusters of Orthologous Groups (COG) database to predict potential genes and provide their functional information. Based on our transcriptome data and published literatures, eight full-length cDNA clones encoding , , , , , , , and genes, involved in galantamine biosynthesis, were identified in . . In order to investigate galantamine biosynthesis in different plant parts of . grown in a growth chamber, gene expression levels were measured through quantitative real-time polymerase chain reaction (qRT-PCR) analysis using these identified genes and galantamine levels were quantified by high-performance liquid chromatography (HPLC) analysis. The qRT-PCR data revealed high expression levels of , , and in the bulbs, and, as expected, we observed higher amounts of galantamine in the bulbs than in the root and leaves. Additionally, a total of 40 hydrophilic metabolites were detected in the different organs using gas-chromatography coupled with time-of-flight mass spectrometry. In particular, a strong positive correlation between galantamine and sucrose, which provides energy for the secondary metabolite biosynthesis, was observed.
属于石蒜科，是一种原产于韩国、中国和日本的球根植物。加兰他敏是石蒜科植物的代表性生物碱，包括……，具有选择性和优势性的乙酰胆碱酯酶抑制作用。尽管……具有经济和药用重要性，但关于……的分子生物学和生化信息相对匮乏。因此，本研究提供了……的功能信息，描述了加兰他敏在各个器官中的生物合成，并提供转录组和代谢数据集，以支持未来研究中对加兰他敏生物合成途径的阐明。重复进行的研究结果显示，在使用CutAdapt修剪原始读段、使用Trinity软件包进行组装以及使用CD-Hit-EST进行聚类后，分别从9,913,869,968和10,162,653,038条原始读段中获得了总共325,609和404,019个单基因。所有组装的单基因都与公共数据库进行了比对，包括美国国立生物技术信息中心（NCBI）的非冗余蛋白质（NR）和核苷酸（Nt）数据库、SWISS-PROT（UniProt）蛋白质序列数据库、拟南芥信息资源库（TAIR）、瑞士蛋白质数据库、基因本体论（GO）和直系同源簇（COG）数据库，以预测潜在基因并提供其功能信息。基于我们的转录组数据和已发表的文献，在……中鉴定出了8个参与加兰他敏生物合成的全长cDNA克隆，分别编码……、……、……、……、……、……、……和……基因。为了研究在生长室中生长的……不同植物部位的加兰他敏生物合成，使用这些鉴定出的基因通过定量实时聚合酶链反应（qRT-PCR）分析测量基因表达水平，并通过高效液相色谱（HPLC）分析对加兰他敏水平进行定量。qRT-PCR数据显示……、……和……在鳞茎中高表达，正如预期的那样，我们观察到鳞茎中的加兰他敏含量高于根和叶。此外，使用气相色谱-飞行时间质谱联用技术在不同器官中总共检测到40种亲水性代谢物。特别是，观察到加兰他敏与蔗糖之间存在强烈的正相关，蔗糖为次生代谢物生物合成提供能量。
