山茶属植物EST-SSR序列分析

楚雄师范学院学报 ›› 2021, Vol. 36 ›› Issue (3): 39-50.

山茶属植物EST-SSR序列分析

董存丽^1,#, 寇国艳^2,#, 罗春梅^3,#, 张燕^4,#, 杨海艳^2,*, 范树国², 陈迁进⁵, 于建功⁵, 绍胜富⁶, 将昌杰⁷, 罗燕英⁷, 邱璐^2,*

1.云南哀牢山国家级自然保护区楚雄管护局,云南楚雄 675000, 中国;
2.楚雄师范学院资源环境与化学学院, 云南楚雄 675000, 中国;
3.楚雄技师学院现代园林工程系,云南楚雄 675000, 中国;
4.楚雄州农业科学院经济作物研究所,云南楚雄 675000, 中国;
5.浙江省金华市林业种苗站,浙江金华 321000;
6.浙江省金华市国际茶花物种园,浙江金华 321000;
7.广西金花茶公园,南宁 530000,中国

收稿日期:2019-11-02 出版日期:2021-05-20 发布日期:2021-07-23

EST-SSR Sequence Analysis of Genus Camellia

Cunli Dong^1,#, Guoyan Kou^2,#, Chunmei Luo^3,#, Yan Zhang^4,#, Haiyan Yang^2,*, Shuguo Fan², Qianjin Chen⁵, Jiangong Yu⁵, Shengfu Shao⁶, Changjie Jiang⁷, Yangying Luo⁷, Lu Qiu^2,*

1.Chuxiong Administration, Ailao Mountain National Nature Reserve of Yunnan, Chuxiong 675000, China;
2. School of Resources, Environment & Chemistry, Chuxiong Normal University, Chuxiong, Yunnan Province 675000;
3. Department of Modern Landscape Engineering,Chuxiong Technician College,Chuxiong,Yunnan 675000,China;
4. Institute of Cash Crops, Chuxiong Academy of Agricultural Sciences, Chuxiong 675000, China;
5. Forestry Seedling Station of Jinhua City, Jinhua 321000, China;
6. Jinhua International Camellia Species Park, Jinhua 321000, China;
7. Golden Flower Tea Park, Guangxi Province, Nanning 530000, China

Received:2019-11-02 Online:2021-05-20 Published:2021-07-23
Contact: *HaiyanYang（1979–）, doctor, associate professor, mainly engaged in plomt pathology, plomtphysiology reserch, E-mail：haiyanyang@cxtc.edu.cn; Lu Qiu（1964–）, professor, mainly engaged in DNA and RNA research, E-mail ： qiulu@cxtc.edu.cn, Tel. 0086-8783120042
About author:#Cunli Dong, senior engineer, mainly engaged in cultivation and protection of forest resources, E-mail： 2824067245@qq.com; Guoyan Kou, mainly engaged in DNA analysis, E-mail：171123518@qq.com; Chunmei Luo, associate professor, mainly engaged in crop growing, E-mail：546367998@qq.com; Yan Zhang, senior researcher, mainly engaged in the research of cash crops, E-mail：2929274868

摘要/Abstract

摘要： 研究通过SSR、EST-SSR分子标记技术对179种山茶属植物进行亲缘关系分析。采用改良的CTAB法提取茶花基因组DNA,用1%的琼脂糖凝胶电泳检测DNA纯度,用2%的琼脂糖凝胶电泳检测PCR扩增产物。扩增产物测序结果显示, EST-SSR序列片段大小为194 bp,序列最长的是崇左金花茶,长度为210 bp, 序列最短为凹脉金花茶,长度为152 bp。在NCBI上寻找同源系列,用Cluster软件进行系列比对,发现不同山茶属植物EST-SSR系列差异较大,其中缺失位点为1752个,突变位点813个。用MEGA6.0软件画系统树,对茶花进行亲缘关系分析,39种茶花聚为红山茶组、金花茶组、糙果茶组、连蕊茶组四组,结果与张宏达形态学分类结果基本一致。结论EST-SSR分子标记用于茶花系统分类及物种鉴定效果较好。

关键词: 山茶属, SSR, EST-SSR, 系统树, 亲缘关系

Abstract: 179 plants of Camellia genus were analyzed using SSR, EST-SSR molecular markers technology to detect their genetic relationship. DNAs of the 179 Camellia plants were extracted by improved CTAB, then tested for the quality of DNAs by way of 1 percent of agarose gel electrophoresis, and tested for PCR amplification by way of 2 percent of agarose gel electrophoresis. The results of amplification products show, the EST-SSR sequence fragment size is 194bp, the longest being Chongzhu （C. theopsis） of 210bp and shortest being Aomai （C. impressinervis） of 152bp. Referring to the homologous series in NCBI and compared using Cluster X software, great differences were found in the EST-SSRs of different camellia plants, with 1752 deletion loci and 813 mutation sites. Phylogenetic trees were constructed using MEGA 6.0 to analyze their genetic relationship, 39 camellia plants were classified into four groups, i.e. Sect. Camellia, Sect. Chrysantha, Sect. Furfuracea and Sect. Theopsis. The results are basically in consistency with Zhang Hongda morphological classification. This indicates that EST-SSR molecular marker is effective in phylogenetic classification and identification of Camellia.

Key words: Genus Camellia, SSR, EST-SSR, phylogenetic tree, genetic relationship

中图分类号:

Q949.758.4

董存丽, 寇国艳, 罗春梅, 张燕, 杨海艳, 范树国, 陈迁进, 于建功, 绍胜富, 将昌杰, 罗燕英, 邱璐. 山茶属植物EST-SSR序列分析[J]. 楚雄师范学院学报, 2021, 36(3): 39-50.

Cunli Dong, Guoyan Kou, Chunmei Luo, Yan Zhang, Haiyan Yang, Shuguo Fan, Qianjin Chen, Jiangong Yu, Shengfu Shao, Changjie Jiang, Yangying Luo, Lu Qiu. EST-SSR Sequence Analysis of Genus Camellia[J]. Journal of Chuxiong Normal University, 2021, 36(3): 39-50.

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