基于SSR分子标记的广西德保县和隆林县野生古茶树聚类分析
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摘要
【目的】对广西德保县和隆林县野生古茶树及其他省份名优栽培种茶树进行聚类分析,为野生古茶树种质资源保护、良种选育及创新利用提供理论参考。【方法】利用筛选出的15对SSR引物对9份广西德保县和隆林县的野生古茶树种质和15份名优栽培种茶树种质进行聚类分析,并计算遗传相似系数。【结果】筛选出的15对引物从德保04和德保05未扩增出任何条带,但从其余22份茶树样品中共扩增出183条谱带,其中CS1引物扩增出多态性谱带最多,为19条,最少为CS13引物,仅7条,平均每对引物扩增出多态谱带12.26条,多态率为96.17%。22份茶树种质资源可聚为两大类群,其中德保县和隆林县的野生古茶树聚在Ⅰ类群,栽培种茶树聚在Ⅱ类群。根据遗传相似系数,也可获得上述相同的分类结果。茶树品种间的遗传相似系数为0.48～0.84,平均遗传相似系数为0.67,其中,野生古茶树种质资源间的遗传相似系数为0.68～0.84;栽培种茶树间的遗传相似系数为0.62～0.84;野生古茶树与栽培种茶树的遗传相似系数为0.48～0.77。仅德保06与其他野生古茶树(隆林42和德保01)遗传相似系数小于0.70;栽培种茶树与野生古茶树相比,仅黄观音与野生古茶树(德保01和德保G02)遗传相似系数大于0.70。【结论】广西德保县和隆林县的野生古茶树种质起源相近,与我国其他各省份的名优栽培种茶树亲缘关系较远,存在相远近亲和相近远亲共存现象,进一步证实广西德保县野生古茶树为五柱茶系的厚轴茶,推测桂西北山区广泛分布山茶属原始种系。
【Objective】Clustering analysis for wild ancient tea germplasm resources in Debao County and Longlin County of Guangxi and cultivated teas was conducted to provide scientific basis for protection of wild ancient tea germplasm resources and fine tea germplasms breeding and innovation and application of them.【Method】Fifteen pairs of SSR primers were used to conduct clustering analysis for nine wild ancient tea germplasm resources in Debao County and Longlin County,Guangxi and fifteen fine cultivated tea germplasm resources,then the genetic similarity coefficient was calculated.【Result】No bands were amplified from Debao 04 and Debao 05 by the selected 15 pairs of primers,but 183 bands were amplified from the rest 22 tea germplasms. CS1 primers amplified the most polymorphism bands(19),and CS13 primers amplified the least polymorphism bands(7). Each pair of primers amplified 12.26 polymorphism bands on avera-ge with polymorphism rate of 96.17%. The 22 tea samples could be clustered into two groups. The first group included all the wild ancient tea germplasm resources in Debao County and Longlin County while the second group included all the cultivated teas. The genetic similarity coefficient also indicated the same results. The genetic similarity coefficient of tea germplasm resources ranged from 0.48 to 0.84. The average of genetic similarity coefficient was 0.67. The similarity coefficients among the wild ancient tea tree population were concentrated in 0.68-0.84,while similarity coefficients among the cultivated teas were 0.62-0.84. The genetic similarity coefficients between wild ancient teas and cultivated teas were 0.48-0.77. Only the similarity coefficient of Debao 06 and other wild ancient teas(Longlin 42 and Debao 01)was less than 0.70.In comparison between wild ancient teas and cultivated teas,only the genetic similarity coefficient between Huangguanyin and wild ancient teas(Debao 01 and Debao 02)was greater than 0.70.【Conclusion】The germplasm origin of wild ancient tea trees in Debao County and Longlin County is close,but they are far with the cultivated teas in other places of China. The germplasms from distant places may be related,and at the same time,the germplasms in the close places may be genetically far away. This further testifies that Debao wild ancient tea blongs to Camellia crassicolumna in C. pentastyla.It is inferred that there are original lines of Camellia widely distributed in north-west mountainous regions in Guangxi.
【Objective】Clustering analysis for wild ancient tea germplasm resources in Debao County and Longlin County of Guangxi and cultivated teas was conducted to provide scientific basis for protection of wild ancient tea germplasm resources and fine tea germplasms breeding and innovation and application of them.【Method】Fifteen pairs of SSR primers were used to conduct clustering analysis for nine wild ancient tea germplasm resources in Debao County and Longlin County,Guangxi and fifteen fine cultivated tea germplasm resources,then the genetic similarity coefficient was calculated.【Result】No bands were amplified from Debao 04 and Debao 05 by the selected 15 pairs of primers,but 183 bands were amplified from the rest 22 tea germplasms. CS1 primers amplified the most polymorphism bands(19),and CS13 primers amplified the least polymorphism bands(7). Each pair of primers amplified 12.26 polymorphism bands on avera-ge with polymorphism rate of 96.17%. The 22 tea samples could be clustered into two groups. The first group included all the wild ancient tea germplasm resources in Debao County and Longlin County while the second group included all the cultivated teas. The genetic similarity coefficient also indicated the same results. The genetic similarity coefficient of tea germplasm resources ranged from 0.48 to 0.84. The average of genetic similarity coefficient was 0.67. The similarity coefficients among the wild ancient tea tree population were concentrated in 0.68-0.84,while similarity coefficients among the cultivated teas were 0.62-0.84. The genetic similarity coefficients between wild ancient teas and cultivated teas were 0.48-0.77. Only the similarity coefficient of Debao 06 and other wild ancient teas(Longlin 42 and Debao 01)was less than 0.70.In comparison between wild ancient teas and cultivated teas,only the genetic similarity coefficient between Huangguanyin and wild ancient teas(Debao 01 and Debao 02)was greater than 0.70.【Conclusion】The germplasm origin of wild ancient tea trees in Debao County and Longlin County is close,but they are far with the cultivated teas in other places of China. The germplasms from distant places may be related,and at the same time,the germplasms in the close places may be genetically far away. This further testifies that Debao wild ancient tea blongs to Camellia crassicolumna in C. pentastyla.It is inferred that there are original lines of Camellia widely distributed in north-west mountainous regions in Guangxi.
引文
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赖建红,吕杨俊,张海华,蒋玉兰.2016.安吉白茶资源全价利用现状与思考[J].中国茶叶加工,(2):29-32.[Lai J H,LüY J,Zhang H H,Jiang Y L.2016.Current situation and thoughts on Anji Baicha tea resources’s comprehensive utilization[J].Chinese Tea Processing,(2):29-32.]
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蓝燕,邱勇娟,叶靖平,陈新强,李朝昌.2017.广西德保县野生古茶树资源调查初报[J].农业科技通讯,(7):359-360.[Lan Y,Qiu Y J,Ye J P,Chen X Q,Li C C.2017.A preliminary investigation of wild ancient tea tree resources in Debao County,Guangxi[J].Agro-technical Communication,(7):359-360.]
李朝昌,蒋漓生.2018a.广西野生茶树图片集[M].桂林:漓江出版社.[Li C C,Jiang L S.2018a.Collection of Guangxi Wild Tea Tree Picture[M].Gulin:Lijiang Press.]
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刘彤,葛智文,陈涛林,杨雪梅,罗军武.2015.柳州九万山野生茶树种质资源遗传多样性的ISSR分析[J].南方农业学报,46(11):1943-1948.[Liu T,Ge Z W,Chen T L,Yang X M,Luo J W.2015.Genetic diversity of wild tea germplasm resources in Liuzhou Jiuwan Mountain revealed by ISSR markers[J].Journal of Southern Agriculture,46(11):1943-1948.]
韦柳花,罗小梅,邓慧群.2017.广西野生茶树种质资源研究进展[J].现代农业科技,(15):51-52.[Wei L H,Luo XM,Deng H Q.2017.Research progress on germplasm resources of wild tea in Guangxi[J].Modern Agricultural Science and Technology,(15):51-52.]
姚明哲.2009.利用ISSR和EST-SSR标记研究中国茶树资源的遗传多样性和遗传结构[D].杭州:浙江大学.[Yao M Z.2009.Studies on genetic diversity and structure of tea germplasm in China based on ISSR and EST-SSRmarkers[D].Hangzhou:Zhejiang University.]
张宏达.1984.茶叶植物资源的订正[J].中山大学学报,(1):1-12.[Zhang H D.1984.Correction of tea plant resources[J].Journal of Sun Yat-sen University,(1):1-12.]
张文驹,戎俊,韦朝领,高连明,陈家宽.2018.栽培茶树的驯化起源与传播[J].生物多样性,26(4):357-372.[Zhang W J,Rong J,Wei C L,Gao L M,Chen J K.2018.Domestication origin and spread of cultivated tea plants[J].Biodiversity Science,26(4):357-372.]
张羽,李佼,席彦军,李秀峰,徐凯明.2017.3种分子标记在分析茶树种质资源遗传多样性上的比较[J].分子植物育种,15(4):1553-1562.[Zhang Y,Li J,Xi Y J,Li X F,Xu K M.2017.Comparative analysis of three molecular markers on germplasm resources genetic diversity in tea plant[J].Molecular Plant Breeding,15(4):1553-1562.]
诸葛天秋,李朝昌,邓慧群,罗跃新,陈新强.2015.广西野生茶树资源调查[J].安徽农业科学,43(29):1-3.[Zhuge T Q,Li C C,Deng H Q,Luo X Q,Chen X Q.2015.Investigation on Guangxi wild tea resources[J].Journal of Anhui Agricultural Sciences,43(29):1-3.]
左志明,陈亮,虞富莲.1994.桂西山区茶树种质资源考察初报[J].广西农业科学,(2):63-65.[Zuo Z M,Chen L,Yu F L.1994.Preliminary report on the investigation of tea germplasm resources in the mountainous area of western Guangxi[J].Guangxi Agricultural Science,(2):63-65.]
陈莹玉,袁思思,吴春兰,赖幸菲,赵文芳,朱燕,杨家干,黄亚辉.2016.广西金秀野生茶遗传多样性及分子指纹图谱研究[J].广东农业科学,43(3):60-65.[Chen Y Y,Yuan S S,Wu C L,Lai X F,Zhao W F,Zhu Y,Yang J G,Huang Y H.2016.Molecular fingerprint and diversity analysis of wild tea trees in Jinxiu County,Guangxi Province[J].Guangdong Agricultural Sciences,43(3):60-65.]
赖建红,吕杨俊,张海华,蒋玉兰.2016.安吉白茶资源全价利用现状与思考[J].中国茶叶加工,(2):29-32.[Lai J H,LüY J,Zhang H H,Jiang Y L.2016.Current situation and thoughts on Anji Baicha tea resources’s comprehensive utilization[J].Chinese Tea Processing,(2):29-32.]
赖兆荣,龙启,邱勇娟,韦柳花,李国政.2015.广西姑辽野生茶树种质资源系统选育优异单株研究[J].现代农业科技,(4):53-55.[Lai Z R,Long Q,Qiu Y J,Wei L H,Li G Z.2015.Study on single plant selection in Guliao wild tea plant germplasm resources in Guangxi[J].Modern Agricultural Science and Technology,(4):53-55.]
蓝燕,邱勇娟,叶靖平,陈新强,李朝昌.2017.广西德保县野生古茶树资源调查初报[J].农业科技通讯,(7):359-360.[Lan Y,Qiu Y J,Ye J P,Chen X Q,Li C C.2017.A preliminary investigation of wild ancient tea tree resources in Debao County,Guangxi[J].Agro-technical Communication,(7):359-360.]
李朝昌,蒋漓生.2018a.广西野生茶树图片集[M].桂林:漓江出版社.[Li C C,Jiang L S.2018a.Collection of Guangxi Wild Tea Tree Picture[M].Gulin:Lijiang Press.]
李朝昌,蒋漓生.2018b.广西野生茶树种质资源集锦[M].桂林:漓江出版社.[Li C C,Jiang L S.2018b.Collection of Guangxi Wild Tea Tree Germplasm Resources[M].Guilin:Lijiang Press.]
刘彤,葛智文,陈涛林,杨雪梅,罗军武.2015.柳州九万山野生茶树种质资源遗传多样性的ISSR分析[J].南方农业学报,46(11):1943-1948.[Liu T,Ge Z W,Chen T L,Yang X M,Luo J W.2015.Genetic diversity of wild tea germplasm resources in Liuzhou Jiuwan Mountain revealed by ISSR markers[J].Journal of Southern Agriculture,46(11):1943-1948.]
韦柳花,罗小梅,邓慧群.2017.广西野生茶树种质资源研究进展[J].现代农业科技,(15):51-52.[Wei L H,Luo XM,Deng H Q.2017.Research progress on germplasm resources of wild tea in Guangxi[J].Modern Agricultural Science and Technology,(15):51-52.]
姚明哲.2009.利用ISSR和EST-SSR标记研究中国茶树资源的遗传多样性和遗传结构[D].杭州:浙江大学.[Yao M Z.2009.Studies on genetic diversity and structure of tea germplasm in China based on ISSR and EST-SSRmarkers[D].Hangzhou:Zhejiang University.]
张宏达.1984.茶叶植物资源的订正[J].中山大学学报,(1):1-12.[Zhang H D.1984.Correction of tea plant resources[J].Journal of Sun Yat-sen University,(1):1-12.]
张文驹,戎俊,韦朝领,高连明,陈家宽.2018.栽培茶树的驯化起源与传播[J].生物多样性,26(4):357-372.[Zhang W J,Rong J,Wei C L,Gao L M,Chen J K.2018.Domestication origin and spread of cultivated tea plants[J].Biodiversity Science,26(4):357-372.]
张羽,李佼,席彦军,李秀峰,徐凯明.2017.3种分子标记在分析茶树种质资源遗传多样性上的比较[J].分子植物育种,15(4):1553-1562.[Zhang Y,Li J,Xi Y J,Li X F,Xu K M.2017.Comparative analysis of three molecular markers on germplasm resources genetic diversity in tea plant[J].Molecular Plant Breeding,15(4):1553-1562.]
诸葛天秋,李朝昌,邓慧群,罗跃新,陈新强.2015.广西野生茶树资源调查[J].安徽农业科学,43(29):1-3.[Zhuge T Q,Li C C,Deng H Q,Luo X Q,Chen X Q.2015.Investigation on Guangxi wild tea resources[J].Journal of Anhui Agricultural Sciences,43(29):1-3.]
左志明,陈亮,虞富莲.1994.桂西山区茶树种质资源考察初报[J].广西农业科学,(2):63-65.[Zuo Z M,Chen L,Yu F L.1994.Preliminary report on the investigation of tea germplasm resources in the mountainous area of western Guangxi[J].Guangxi Agricultural Science,(2):63-65.]