目的 解析怀玉山产三叶青Tetrastigma hemsleyanum叶绿体基因组信息序列特征和确定其在崖爬藤属的系统位置。方法 用Illumina高通量测序平台NovaSeq6000进行测序获得怀玉山产三叶青叶绿体基因组序列，借助GeSeq、tRNAscan-SE、MISA、VISTA tools、DNADnaSP6.0、JSHYCloud、CodonW1.4.2、Pasteur Galaxy、mafft 7.0、fasttree 2.1.10等生物信息学工具进行序列分析、密码子偏好分析、崖爬藤属基因组比较分析和系统发育研究。结果 怀玉山产三叶青Tetrastigma hemsleyanum叶绿体基因组为共价闭合双链环状分子，长160 165 bp，包含1个大单拷贝区（large single copy region，LSC）、1个反向重复区a（inverted repeat region a，IRa）、1个反向重复区b（inverted repeat region b，IRb）和1个小单拷贝区（small single copy region，SSC）；怀玉山产三叶青叶绿体基因组注释到光合作用基因、自我复制基因、其他基因和未知功能基因4类共133个基因，包括 88个CDS基因、37个tRNA基因和8个rRNA基因；怀玉山产三叶青叶绿体基因组检测到63个SSR位点，其中单碱基重复56个，双碱基重复7个；怀玉山产三叶青叶绿体基因组核苷酸多样性的变化范围为0～0.234 18，基因间隔区trnS-GCU-trnG-UCC、rps16-trnG-UCC以及基因ycf1变异率最高。怀玉山产三叶青叶绿体基因组密码子偏好以A或U结尾，具有较强的偏好性。突变对密码子偏好性的影响较强，而其他因素如自然选择对密码子的偏好性影响较小。怀玉山产三叶青叶绿体基因组有31个高频密码子，16个以U结尾，13个以A结尾，2个以G结尾，其中13个为最优密码子。怀玉山产三叶青与福建产三叶青（MW375708）和广西产三叶青（MW375709）亲缘关系较近，与浙江产三叶青（MT827073）、浙江产三叶青（MW375707）、浙江产三叶青KT033563（NC_029339）、四川产三叶青（MW375710）和浙江产三叶青（MW375711）亲缘关系较远。结论 三叶青之间的亲缘关系与地理位置相关性不大。首次采用生物信息学分析方法对怀玉山产三叶青叶绿体基因组进行了全面、深度解析，结果将为崖爬藤属药用植物的遗传多样性分析以及新品系的遗传育种研究等提供理论依据。

Objective To analyze the genomic information sequence characteristics of the chloroplast of Tetrastigma hemsleyanum from Huaiyu Mountain and determine its systematic position in the genus of Tetrastigma. Methods The chloroplast genome sequence of T. hemsleyanum from Huaiyu Mountain was obtained by sequencing with Illumina high throughput sequencing platform NovaSeq6000, and bioinformatics tools such as GeSeq, tRNAscan SE, MISA, VISTA tools, DNADnaSP6.0, JSHYCloud, CodonW1.4.2, Pasteur Galaxy, Mafft 7.0, fasttree 2.1.10 were used for sequence analysis, codon preference analysis, genome comparative analysis and phylogenetic research of Tetrastigma. Results The chloroplast genome of T. hemsleyanum from Huaiyu Mountain is a covalently closed double stranded ring molecule with a length of 160 165 bp, including 1 LSC, 1 IRa, 1 IRb and 1 SSC; the chloroplast genome of T. hemsleyanum from Huaiyu Mountain has been annotated with 133 genes in four categories: photosynthesis genes, self-replication genes, other genes and unknown functional genes, including 88 CDS genes, 37 tRNA genes and eight rRNA genes; Sixty three SSR loci were detected in the chloroplast genome of T. hemsleyanum from Huaiyu Mountain, including 56 single base repeats and seven double base repeats; The variation range of nucleotide diversity of the chloroplast genome of T. hemsleyanum from Huaiyu Mountain was 0—0.234 18, and the variation rates of trnS-GCU-trnG UCC, rps16-trnG UCC and gene ycf1 were the highest. The codon of chloroplast genome of T. hemsleyanum from Huaiyu Mountain had a strong preference for ending in A or U. Mutation had a strong impact on codon preference, while other factors such as natural selection had a small impact on codon preference. The chloroplast genome of T. hemsleyanum from Huaiyu Mountain had 31 high-frequency codons, 16 ending in U, 13 ending in A, and two ending in G, of which 13 were the optimal ones. T. hemsleyanum from Huaiyu Mountain was close to T. hemsleyanum from Fujian MW375708 and T. hemsleyanum from Guangxi MW375709, and far from T. hemsleyanum from Zhejiang MT827073, T. hemsleyanum from Zhejiang MW375707, T. hemsleyanum from Zhejiang KT033563 (NC_029339), T. hemsleyanum from Sichuan MW375710 and T. hemsleyanum from Zhejiang MW375711, which showed that there was little correlation between the genetic relationship and the geographical location. Conclusion The genetic relationship between T. hemsleyanum is not closely related to its geographical location. In this study, the chloroplast genome of T. hemsleyanum from Huaiyu Mountain is analyzed comprehensively and deeply for the first time by using bioinformatics analysis method, which will provide theoretical basis for genetic diversity analysis of medicinal plants and genetic breeding research of new strains of Tetrastigma.

R286.12

国家自然科学基金资助项目（31960079）；2022年上饶市科技专项项目（饶科发[2023]5号社发类）（2022A008）；江西省科技厅重点研发计划一般项目（20202BBG73010）；江西省教育厅科学技术研究项目（GJJ201704）；上饶市科技局平台载体建设项目（2020J001）