茶组植物的分子系统学研究
|
摘要
茶组Sect．Thea(L．)Dyer属于山茶属Genus Camellia Linn．、山茶科Theaceae。除茶‘C．sinensis(L．)O．Kuntze’为广布种外，其它种几乎全产于中国南部与西南部，它是山茶属中最有价值的一个组，而茶又是茶组中最重要的一种。作者对茶树基因组DNA提取方法、茶树(种内)资源遗传多样性、茶树资源的分子标记鉴别、茶组分子系统学和形态系统学等进行了较系统深入的研究，获得以下重要结果。
1、在前人研究基础上，建立了茶树基因组DNA的SDS-异丙醇提取法。该法提取缓冲液使细胞维持一定的渗透压，研磨时使细胞核基本保持完整；在细胞核被裂解之前去除细胞质中的茶多酚、大部分蛋白质和RNA；而后用SDS裂解细胞核，异丙醇或乙醇沉淀DNA，这样能经济、快速和有效地从富含茶多酚、茶多糖等次生物质的茶树新梢中提取基因组DNA。基因组DNA平均得率为527μg／g鲜重，经鉴定适合进行限制性酶切和RAPD扩增。基因组DNA质量以新梢最好，干梢次之；液氮(-196℃)冷冻保存好于-20℃保存。应用硅胶脱水干燥成功保存茶树新梢，对解决野外远距离采样具有实践意义。
2、从100个随机引物中筛选出20个用于对我国15份茶树(种内)资源遗传多样性的RAPD扩增，共得到1050个位点，平均52.5个／引物，70个／资源。在全部的137条谱带中，8条是共有的(占5.8％)，多态性程度达94.2％，资源间的遗传距离在0.16-0.62之间，平均为0.37，遗传多态性和遗传距离远比肯尼亚、韩国、日本等大，从DNA分子水平上更加证实了中国是茶树的原产地和起源中心。引物的多态性频度在0.07至0.93之间，总平均为0.47，属于比较高的多态性频度，其中引物S201多态性频度最小，平均为0.24；引物207最大，平均为0.83。类平均法聚类结果显示，可划分为3个类群，同时探讨了类群内和类群间的亲缘关系。
3、对原产于云南等地的24份茶树资源进行分子标记鉴别研究，表明RAPD标记在鉴别茶树资源方面非常有效。有3种独立的方法：1)特殊的标记；2)特异的谱带类型；3)不同引物提供谱带类型的组合，可用于茶树资源的分子标记鉴别。OPO-13等12个引物共存在15个和缺失3个特异标记，可以鉴别其中14份资源；引物OPO-13扩增的14种谱带类型可以鉴别10份资源；2-3个引物谱带类型的组合分别可以鉴别15、17和20份资源。利用最少数量引物获得最大鉴别能力，对
资源的分子鉴别尤为重要。OPO－13，OPG－12，OPO－18和 OPA－13等 4个引物带
型的组合可以鉴别所有24份资源，包括形态和生化成分上几乎没有差异的2株毗
邻大茶树。
4、从 Operon公司的 61个引物中，筛选出 15个用于 24种、变种茶组植物的
RAPD扩增，在所扩增得到的107条可重现谱带中（平均7＊条污物）有102条（平
均6．8条尺物）是多态的，多态性程度是95．3％。引物多态性频度在0刀个096之间，
总平均为0．30，其中OPO－19多态性频度最小，平均为0．16；OPG－15最大，平均
为0石0。利用ipD标记构建的分子系统树和SHMM主成分分析均可将茶组植物
分为“五室类群”和“三室类群”两大类。在UPGMA分子系统树上，两大类下
可分别再分3个和2个亚类群，这和茶组植物的细胞学、化学和数学分类法基本
一致。同时从遗传距离上探讨了一些种的亲缘关系和分子进化。
综上所述，RAPD标记不仅适合于茶树种内水平，也同样适合于组内种间水平，
为茶树及其近缘植物遗传多样性、亲缘关系和分类鉴别提供了新颖可靠的分子学
方法和证据。
5、通过对野生大茶树居群、其它茶树资源等的系统研究以及茶组分子系统学
分析，结合前人的茶组分类研究，把张宏达分类系统的42种4变种茶组植物进行
归并，对其形态学分类系统进行了修订。该系统以张氏系统“系”的划分为基础，
以子房室数、花柱分裂数和子房茸毛为主要依据，结合中轴大小、果皮厚度、花
冠大小、花尊茸毛及树型、枝叶等的形态特征，将它们归并为大厂茶Camellia
tQCh。nge皿Is下．C．Zh。ig、大理茶 C toll删is（W W．S＊ith）MCIChi。f、厚轴茶 C
c。icolumna Chang、秃房茶 C gythnogya Chang和茶 C s。enslA（L．）O．KntZe共
5种，在茶下分普洱茶C sin删is var。am。（Masters）im。a、白毛茶C sin。S0
var Pubilimba Chang 2变种。野生型茶树主要属于大厂茶、厚轴茶、大理茶、秃房
茶。栽培型茶树主要属于茶及普洱茶、白毛茶等变种。除茶为广布种外，其余4
种、2变种主要分布在我国云南、广西和贵州等地。茶组的系统演化途径可分成子
房多毛和无毛二条路线，由子房5室向3室，乔木向灌木，大花多瓣向小花少瓣
演化。
Sect. Thea (L.) Dyer was a section in the genus Camellia L., Theaceae. Excluding tea plant (C. sinensis), which was a widely spread species both in China and the world, it was centralized distributed in the south and southwest China. This was an important section in Camellia. Tea plant was the most important cash crop in the section. In this dissertation, a novel genomic DNA isolation method, the genetic polymorphism and molecular discrimination of tea plant, the molecular and morphological systematics of section Thea were systematically investigated. The followings were main results.
1. The isolation of genomic DNA was the first step of molecular markers research. A SDS-iso-propanol method suitable for tea plant, which was plentiful of tea polyphenols, had been developed using a modification of Chen Darning's method from different sample storage conditions such as fresh, dry and frozen shoots. It was a quick, easy, economical and effective method. The tactics were as follows: Before the cell nuclear membranes were decomposed, the tea polyphenols and proteins etc.were removed. This step was critical for preventing polyphenol to be oxidized into quinone. Then the nucleuses were splitted by SDS, DNA was precipitated by iso-propanol or alcohol. The average DNA yields were 527 u g/g with fresh base. The DNA quality was fresh > dry ones; stored in liquid N2>in -20 癈. All of them were suitable for RAPD and RFLP analyses. The successful use of silica to dry tea shoots was meaningful for wild fields and long-distance sample collection.
2. Twenty primers, which could amplify polymorphism in 15 Chinese tea clonal germplasms, were selected from 100 arbitrary 10-mer primers. A total of 1050 loci with an average 52.5 loci per primer and 70 loci per germplasm were amplified. In the total 137 amplified bands, 129 were polymorphic, corresponding to 94.2%. The genetic distances were 0.16 to 0.62, on average 0.37. The genetic polymorphism and distance were much higher and wider than those of Kenya, South Korea and Japan. The polymorphic frequency was between 0.07 and 0.93, on an average of 0.47. Primer 201 was the lowest, average 0.24; meanwhile, S207 average 0.83, the highest. The result of UPGMA grouped the 15 tea germplasms into 3 groups. The intra-group and inter-group relationship was analyzed.
3. The discrimination of 24 wild tea germplasm resources using DNA markers was conducted. The result showed that RAPD marker was a very effective tool and robust method in tea germplasm discrimination. There were 3 independent ways to identify the tea germplasms, a) unique RAPD markers, b) unique band patterns and c) a combination of the band patterns provided by different primers. The presence of 15 unique RAPD markers and the absence of 3 unique markers obtained from 12 primers made it possible to identify 14 tea germplasms. Using the unique band patterns of primer OPO-13 could identify 10 tea germplasms. 15, 17 and 20 germplasms could be identified using the band patterns combination of two or three primers, respectively. It was of much importance using minimum primers to obtain the maximum discrimination capacity. All the 24 tea germplasms could be entirely discriminated by the band patterns combination of primer OPO-13, OPO-12, OPG-18 and OPA-13, including two wild tea
plants of very similar morphological characteristics and chemical components.
4. The genetic polymorphism and molecular systematics of 24 species and varieties
in section Thea were investigated. Fifteen arbitrary 10-mer primers were selected from the 61 screened. A total of 102 polymorphic bands (6.8 polymorphisms/ primer) out of 107 reproducible products (7.1 fragments/primer) were amplified from the selected 15 primers, corresponding to 95.3% polymorphism of the amplification bands. The relative polymorphic frequency ranged from 0.04 to 0.96. However, the general relative frequency of polymorphic was low with an average of 0.30, varying from 0.16 to 0.60. Molecular phylogenetic dendrogram of section Thea was constructed using UPGMA a
1、在前人研究基础上，建立了茶树基因组DNA的SDS-异丙醇提取法。该法提取缓冲液使细胞维持一定的渗透压，研磨时使细胞核基本保持完整；在细胞核被裂解之前去除细胞质中的茶多酚、大部分蛋白质和RNA；而后用SDS裂解细胞核，异丙醇或乙醇沉淀DNA，这样能经济、快速和有效地从富含茶多酚、茶多糖等次生物质的茶树新梢中提取基因组DNA。基因组DNA平均得率为527μg／g鲜重，经鉴定适合进行限制性酶切和RAPD扩增。基因组DNA质量以新梢最好，干梢次之；液氮(-196℃)冷冻保存好于-20℃保存。应用硅胶脱水干燥成功保存茶树新梢，对解决野外远距离采样具有实践意义。
2、从100个随机引物中筛选出20个用于对我国15份茶树(种内)资源遗传多样性的RAPD扩增，共得到1050个位点，平均52.5个／引物，70个／资源。在全部的137条谱带中，8条是共有的(占5.8％)，多态性程度达94.2％，资源间的遗传距离在0.16-0.62之间，平均为0.37，遗传多态性和遗传距离远比肯尼亚、韩国、日本等大，从DNA分子水平上更加证实了中国是茶树的原产地和起源中心。引物的多态性频度在0.07至0.93之间，总平均为0.47，属于比较高的多态性频度，其中引物S201多态性频度最小，平均为0.24；引物207最大，平均为0.83。类平均法聚类结果显示，可划分为3个类群，同时探讨了类群内和类群间的亲缘关系。
3、对原产于云南等地的24份茶树资源进行分子标记鉴别研究，表明RAPD标记在鉴别茶树资源方面非常有效。有3种独立的方法：1)特殊的标记；2)特异的谱带类型；3)不同引物提供谱带类型的组合，可用于茶树资源的分子标记鉴别。OPO-13等12个引物共存在15个和缺失3个特异标记，可以鉴别其中14份资源；引物OPO-13扩增的14种谱带类型可以鉴别10份资源；2-3个引物谱带类型的组合分别可以鉴别15、17和20份资源。利用最少数量引物获得最大鉴别能力，对
资源的分子鉴别尤为重要。OPO－13，OPG－12，OPO－18和 OPA－13等 4个引物带
型的组合可以鉴别所有24份资源，包括形态和生化成分上几乎没有差异的2株毗
邻大茶树。
4、从 Operon公司的 61个引物中，筛选出 15个用于 24种、变种茶组植物的
RAPD扩增，在所扩增得到的107条可重现谱带中（平均7＊条污物）有102条（平
均6．8条尺物）是多态的，多态性程度是95．3％。引物多态性频度在0刀个096之间，
总平均为0．30，其中OPO－19多态性频度最小，平均为0．16；OPG－15最大，平均
为0石0。利用ipD标记构建的分子系统树和SHMM主成分分析均可将茶组植物
分为“五室类群”和“三室类群”两大类。在UPGMA分子系统树上，两大类下
可分别再分3个和2个亚类群，这和茶组植物的细胞学、化学和数学分类法基本
一致。同时从遗传距离上探讨了一些种的亲缘关系和分子进化。
综上所述，RAPD标记不仅适合于茶树种内水平，也同样适合于组内种间水平，
为茶树及其近缘植物遗传多样性、亲缘关系和分类鉴别提供了新颖可靠的分子学
方法和证据。
5、通过对野生大茶树居群、其它茶树资源等的系统研究以及茶组分子系统学
分析，结合前人的茶组分类研究，把张宏达分类系统的42种4变种茶组植物进行
归并，对其形态学分类系统进行了修订。该系统以张氏系统“系”的划分为基础，
以子房室数、花柱分裂数和子房茸毛为主要依据，结合中轴大小、果皮厚度、花
冠大小、花尊茸毛及树型、枝叶等的形态特征，将它们归并为大厂茶Camellia
tQCh。nge皿Is下．C．Zh。ig、大理茶 C toll删is（W W．S＊ith）MCIChi。f、厚轴茶 C
c。icolumna Chang、秃房茶 C gythnogya Chang和茶 C s。enslA（L．）O．KntZe共
5种，在茶下分普洱茶C sin删is var。am。（Masters）im。a、白毛茶C sin。S0
var Pubilimba Chang 2变种。野生型茶树主要属于大厂茶、厚轴茶、大理茶、秃房
茶。栽培型茶树主要属于茶及普洱茶、白毛茶等变种。除茶为广布种外，其余4
种、2变种主要分布在我国云南、广西和贵州等地。茶组的系统演化途径可分成子
房多毛和无毛二条路线，由子房5室向3室，乔木向灌木，大花多瓣向小花少瓣
演化。
Sect. Thea (L.) Dyer was a section in the genus Camellia L., Theaceae. Excluding tea plant (C. sinensis), which was a widely spread species both in China and the world, it was centralized distributed in the south and southwest China. This was an important section in Camellia. Tea plant was the most important cash crop in the section. In this dissertation, a novel genomic DNA isolation method, the genetic polymorphism and molecular discrimination of tea plant, the molecular and morphological systematics of section Thea were systematically investigated. The followings were main results.
1. The isolation of genomic DNA was the first step of molecular markers research. A SDS-iso-propanol method suitable for tea plant, which was plentiful of tea polyphenols, had been developed using a modification of Chen Darning's method from different sample storage conditions such as fresh, dry and frozen shoots. It was a quick, easy, economical and effective method. The tactics were as follows: Before the cell nuclear membranes were decomposed, the tea polyphenols and proteins etc.were removed. This step was critical for preventing polyphenol to be oxidized into quinone. Then the nucleuses were splitted by SDS, DNA was precipitated by iso-propanol or alcohol. The average DNA yields were 527 u g/g with fresh base. The DNA quality was fresh > dry ones; stored in liquid N2>in -20 癈. All of them were suitable for RAPD and RFLP analyses. The successful use of silica to dry tea shoots was meaningful for wild fields and long-distance sample collection.
2. Twenty primers, which could amplify polymorphism in 15 Chinese tea clonal germplasms, were selected from 100 arbitrary 10-mer primers. A total of 1050 loci with an average 52.5 loci per primer and 70 loci per germplasm were amplified. In the total 137 amplified bands, 129 were polymorphic, corresponding to 94.2%. The genetic distances were 0.16 to 0.62, on average 0.37. The genetic polymorphism and distance were much higher and wider than those of Kenya, South Korea and Japan. The polymorphic frequency was between 0.07 and 0.93, on an average of 0.47. Primer 201 was the lowest, average 0.24; meanwhile, S207 average 0.83, the highest. The result of UPGMA grouped the 15 tea germplasms into 3 groups. The intra-group and inter-group relationship was analyzed.
3. The discrimination of 24 wild tea germplasm resources using DNA markers was conducted. The result showed that RAPD marker was a very effective tool and robust method in tea germplasm discrimination. There were 3 independent ways to identify the tea germplasms, a) unique RAPD markers, b) unique band patterns and c) a combination of the band patterns provided by different primers. The presence of 15 unique RAPD markers and the absence of 3 unique markers obtained from 12 primers made it possible to identify 14 tea germplasms. Using the unique band patterns of primer OPO-13 could identify 10 tea germplasms. 15, 17 and 20 germplasms could be identified using the band patterns combination of two or three primers, respectively. It was of much importance using minimum primers to obtain the maximum discrimination capacity. All the 24 tea germplasms could be entirely discriminated by the band patterns combination of primer OPO-13, OPO-12, OPG-18 and OPA-13, including two wild tea
plants of very similar morphological characteristics and chemical components.
4. The genetic polymorphism and molecular systematics of 24 species and varieties
in section Thea were investigated. Fifteen arbitrary 10-mer primers were selected from the 61 screened. A total of 102 polymorphic bands (6.8 polymorphisms/ primer) out of 107 reproducible products (7.1 fragments/primer) were amplified from the selected 15 primers, corresponding to 95.3% polymorphism of the amplification bands. The relative polymorphic frequency ranged from 0.04 to 0.96. However, the general relative frequency of polymorphic was low with an average of 0.30, varying from 0.16 to 0.60. Molecular phylogenetic dendrogram of section Thea was constructed using UPGMA a
引文
1．马小军，汪小全，徐绍玺，肖培根，洪德元．2000，人参不同栽培群体遗传关系的RAPD分析．植物学报，42(6)：587-590
2．王文，凌发瑶，施立明．1994，银额果蝇自然群体中的mtDNA多态性研究(Ⅱ)．银额果蝇的起源和分化．遗传学报，21(4)：263-274
3．王建波，张文驹，陈家宽．1999，核rDNA的ITS序列在被子植物系统与进化研究中的应用．植物分类学报，37(4)：407-416
4．王斌，李松涛，1995，RAPD及其在植物分子生物学研究中的应用．见匡廷云，荆玉祥，李德葆主编．植物分子生物学研究——成就与前景．北京：科学出版社，pp：295-303
5．冯丽春，杨光伟，余茂德、张孝勇、向仲怀．1997，利用RAPD对桑属植物种间亲缘关系的研究．中国农业科学，30(1)：52-56
6．叶创兴．1987，山茶属三新种．中山大学学报(自然科学版)，(1)：17-20
7．刘春林，官春云，李恂．2000，关于植物随机引物扩增多态性DNA标记的可靠性问题．植物生理学通讯，36(1)：56-59
8．向巧萍，向秋云，Liston A，傅立国，傅德志．2000，ITS(nrDNA)片段在冷杉属植物中的长度多态性及其在松科的系统与演化中的应用．植物学报，42(9)：946-951
9．庄晚芳，刘祖生，陈文怀．1981，论茶树变种分类．浙江农业大学学报，7(1)：41-47
10．朱旗，任春梅，洪亚辉，罗军武．1994，茶树叶片DNA提取、纯化与检测．湖南农学院学报，20(2)：114-117
11．吴世安，吕海亮，杨继，饶广远，尤瑞麟，葛颂，钟扬．2000，叶绿体DNA片段的RFLP分析在黄精族系统学研究中的应用．植物分类学报，38(2)：97-110
12．吴美贞．1994，Studies on genetic relationship among the Korean native tea trees and physicochemical properties of its green tea.(韩国)高丽大学校大学院农学科(博士学位论文)，PP：27-29
13．张军丽，王铮峰，李鸣光，王伯荪．2000，植物类群研究中的分子标记及其应用．应用生态学报，11(4)：631-636
14．张宏达．1981a，山茶属植物的系统研究．中山大学学报(自然科学版)论丛，(1)：1-124
15．张宏达．1981b，茶树的系统分类．中山大学学报(自然科学版)，(1)：87-99
16．张宏达．1984，茶叶植物资源的订正．中山大学学报(自然科学版)，(1)：1-12
17．张宏达．1990，中国山茶科植物新种．中山大学学报(自然科学版)，29(2)：85-93
18．张芳赐．1980，云南山茶属的二新种．云南植物研究，2(3)：341-344
19．张芳赐，丁渭然，黄毅，陈兴琰，陈国本，唐明德．1990，云南山茶属的三新种．云南植物研究，12(1)：31-34
20．张芳赐．1994，云南山茶属新变种—邦威茶．见《中国古茶树》，中华茶人联谊会，中国茶叶学会编．上海：上海文化出版社，PP：33-34
21．李光涛，梁涛．1990，中国山茶属4种2变种核型研究．广西植物，10(3)：189-197
22．杜琪珍，李名君，刘维华，王海思．1990，茶组植物的化学分类及数值分类．茶叶科学，10(2)：1-12
23．汪小全，邵喻苹，张大明，洪德元．1996a，银杉遗传多样性的RAPD分析．中国科学(C辑)，(26)：436-441
24．汪小全，邵喻苹，张大明，张志宪，洪德元．1996b，RAPD用于遗传多样性和系统学研究中的问题．植物学报，38(12)：954-962
25．汪小全，洪德元．1997，植物分子系统学近五年的研究进展概况．植物分类学报，35(5)：465-480
26．汪劲武．1984，种子植物分类学．高等教育出版社，PP：1-24
27．邱丽娟，Nelson RL,Vodkin LO.1997，利用RAPD标记鉴定大豆种质．作物学报，23(3)：408-417
28．邹喻苹，汪小全，雷一丁，裴颜龙，张志宪．1994，几种濒危植物及其近缘类群总DNA的提取与鉴定．植物学报，36(7)：528-533
29．邹喻苹，葛颂，王晓东．系统与进化植物学中成分子标记．2001，北京：科学出版社
30．闵天禄．1992，山茶属茶组植物的订正．云南植物研究，14(2)：115-132
31．闵天禄主编．2000，世界山茶属的研究．昆明：云南科技出版社，PP：1-313
32．陈大明，张上隆，金勇丰．1997，一种木本果树基因组DNA提取方法研究．浙江农业大学学报，23(6)：621-624
33．陈亮，杨亚军．1999，茶树分子标记和基因克隆研究进展．茶叶文摘，13(5)：1-6
34．陈亮，杨亚军．2000，RAPD及其在茶树种质资源和遗传育种研究中的应用．海峡两岸茶叶科技学术研讨会．福州4／26-29pp：54-58
35．陈亮，童启庆，高其康，束际林，虞富莲．1997，山茶属8种1变种花粉形态比较．茶叶科学，17(2)：183-188
36．陈亮，虞富莲，童启庆．2000，关于茶组植物分类与演化的讨论．茶叶科学，20(2)：89-94
37．陈亮．1996，茶组植物系统分类学研究现状．茶叶，22(2)：16-19
38．周奕华，陈正华．1999，分子标记在植物学中的应用及前景．武汉植物学研究，17(1)：75-86
39．金惠淑，梁月荣，陆建良．2001，中、韩两国主要茶树品种DNA多态性比较研究．茶叶科学，21(2)：103-107
40．施苏华，唐绍清，陈月琴，屈良鹄，张宏达．1998，11种金花茶植物的RAPD分析及其系统学意义．植物分类学报，36(4)：465-480
41．贾建航，王萍，金德敏，曲雪萍，王倩，李传友，翁曼丽，王斌．2000，RAPD标记在紫菜遗传多样性检测和种质鉴定中的应用．植物学报，42(4)：403-407
42．贾继增．1996，分子标记种质资源鉴定和分子标记育种．中国农业科学，29(4)：1-10
43．郭旺珍，张天真，潘家驹．1996，我国棉花主栽品种的RAPD指纹图谱分析．农业生物技术学报，4(2)：129-134
44．钱韦，葛颂，洪德元．2000，采用RAPD和ISSR标记探讨中国疣粒野生稻的遗传多样性．植物学报，42(7)：741-750
45．梁月荣，田中淳一，武田善行．2000，应用RAPD分子标记分析“晚绿”品种的杂交亲本．茶叶科学，20(1)：22-26
46．梁红健，刘敏，钟志宇，吴应祥，李文彬．1996，中国部分兰花品种RAPD分析．园艺学报，23(4)：365-370
47．梁国鲁，周才琼，林蒙嘉，陈家玉，刘君素．1994，贵州大树茶的核型变异与进化．植物分类学报，32(4)：308-315
48．梁盛业，钟业聪．1981，中国山茶科一个新种．中山大学学报(自然科学版)，(3)：118-119
49．彭英．1992，蛋白质亚基的分离及其与茶组植物的分类．中国茶叶，14(5)：10-11
50．彭隽敏，孔清，徐乃瑜．1995，云南小麦、西藏半野生小麦和普通小麦叶绿体DNA限制性内切酶图谱的研究．遗传，17(6)：4-6
51．程须珍，Yang CY.2001，利用RAPD标记鉴定绿豆组植物种间的亲缘关系．中国农业科学，34(2)：216-218
52．虞富莲．1986，论茶树原产地和起源中心．茶叶科学，6(1)：1-8
53．裴颜龙，邵喻苹，尹蓁，汪小全，张志宪，洪德元．1995，矮牡丹和紫斑牡丹RAPD分析初报．植物分类学报，33(4)：350-356
54．谭永济，陈炳环，虞富莲，王海思，王平盛．1984，中国云南茶树新种和新变种．茶叶科学，4(1)：19-30
55．黎星辉，施兆鹏，刘春林，罗军武，沈程文，龚志华．2001，云南大叶茶与汝城白毛茶杂交后代的RAPD鉴定．茶叶科学，21(2)：99-102
56．黎星辉，施兆鹏，刘春林，罗军武，沈程文，李家贤，方华春．2002，汝城白毛茶与两广主要“白毛茶”居群亲缘关系的RAPD分析．茶叶科学，22(2)：79-82
57．戴思兰，陈俊愉，高荣孚，马江生，李文彬．1996，DNA提纯方法对9种菊属植物RAPD的影响．园艺学报，23(2)：169-174
58. Ainouche ML, Bayer R. 1997, On the origins of the tetraploid Bromus species (section Brornus, Poaceae): insights from internal transcribed spacer sequences of nuclear ribosomal DNA. Genome, 730-743
59. Backmann JS, Soller M. 1983, Restriction fragment length polymorphism in genetic improvement: methodologies, mapping and cost. Theor. Appl. Genet., 67:
35-43
60． Baldwin BG. 1993, Molecular phylogenetics of Calycadenia (Compositae) based on ITS sequences of nuclear nbosomal DNA: chromosomal and morphological evolution reexamined. Amer J Bot, 80: 222-238
61． Baldwin BG, Sanderson MJ, Porter JM, Wojciechowski MF, Campbell CS, Donoghue MJ. 1995, The ITS region of nuclear ribosomal DNA: A valuable source of evidence on angiosperm phylogeny. Ann. Missouri Bot. Card., 247-277
62． Belaj A, Trujilo I, Rosa R, Rallo L, Gimenez MJ. 2001, Polymorphism and discrimination capacity of randomly amplified polymorphic markers in an olive germplasm bank. J. Amer. Soc. Hort. Sci., 126(1) : 64-71
63． Boonprakob U, Byrne D H, Graham C J, Okie W R, Bechman T, Smith B R. 2001, Genetic relationships among cultivated diploid plums and their progenitors as determinated by RAPD markers. J. Amer. Soc. Hort. Sci., 126(4) : 451-461
64． Borthakur S. Mondal TK, Parveen SS, Guha A, Sen P, Borthakur A, Deka PC. 1998, Isolation of chloroplast DNA from tea, Camellia sp. Indian Journal of Experimental Biology, 36: 1165-1167
65． Cabrita LF, Aksoy U, Hepaksoy S Leitao JM. 2001, Suitability of isozyme, RAPD and AFLP markers to access genetic differences and relatedness among fig (Ficus carica L.) clones. Scientia Horticulture, 87: 261-273
66． Cano RJ, Poniar HN, Pieniazek NJ, Acra A, Poinar GO. 1993, Amplification and sequencing of DNA from a 120-135-million-year-old weevil. Nature, 363(10) : 536-538
67． Chen L, Yu FL, Luo LH, Tong QQ. 2001, Morphological classification and phylogenetic evolution of section Thea in the genus Camellia. In: Proceedings of 2001 International Conference on O-Cha (Tea) Science (Session II), pp: 29-32, Oct. 5-8,2001 Shizuoka, Japan
68． Conner PJ, Wood BW. 2001, Identification of pecan germplasms and their genetic relatedness as determined by randomly amplified polymorphic DNA analysis. J.
Amer. Soc. Hort. Sci., 126(4) : 474-480
69． Cornelius PL, Seyedsadr M, Crossa J. 1992, Using the shifted multiplicative model to search for "separability" in crop cultivar trials. Theor. Appl. Genet., 84: 161-172
70． Dellaporta SL, Wood J, Hicks JB. 1983, A plant DNA mini preparation, Version II. Plant Mol. Biol. Rep., 1:19-21
71． Demeke T, Adams R P, Chibbar R. 1992, Potenital taxonomic use of randon amplified polymorphic DNA (RAPD): A case study in Brassica. Theor. Appl. Genet., 84: 990-994
72． Dettori MT, Palombi MA, 2000, Identification of Feijoa Sellowiana Berg accessions by RAPD markers. Scientia Horticulturae, 86: 279-290
73． Doyle JJ, Doyle JL. 1987, A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19: 11-15
74． Elder JR, Turner BJ. 1995, Concerted evolution of repetitive DNA sequences in eukaryotes. Quart. Rev. Biol., 70: 297-319
75． Fatokun CA, Danesh D, Young ND, Stewart EL. 1993, Molecular taxonomic relationships in the genus Vigna based on RFLP analysis. Theor. Appl. Genet., 86: 97-104
76． Federici CT, Fang DQ, Scora RW, Roose ML. 1998, Phylogenetic relationships within the genus citrus (Rutaceae) and related genera as revealed by RFLP and RAPD analysis. Theor. Appl. Genet., 96: 812-822
77． Guillemaut P, Drouard LM. 1992, Isolation of plant DNA: A fast, inexpensive, and reliable methods. Plant Mol. Biol Rep., 10: 60-65
78． Gulsen O, Roose ML. 2001, Chlorplast and nuclear genome analysis of the parenage of lemons. J. Amer. Soc. Hon. Sci., 126(2) : 210-215
79． Hackett CA, Wachira FN, Paul S, Powell W, Waugh R. 2000, Construction of a genetic linkage map for Camellia sinensis (tea). Heredity, 85(4) : 346-355
80． Halward T, Stalker T, LaRue E. 1992, Use of single primer DNA amplification in genetic studies of peanut (Arachis hypogaea L.). Plant mol. Biol., 18: 315-325
81． Hsiao C, Chatterton NJ, Asay KH, Jensen KB. 1995, Phylogenetic relationships of the mono species of wheat tribe, Triticeae (Poaceae) based on nuclear rDNA (ITS) sequence. Theor. Appl. Genet., 90: 389-398
82． Hu J, Quiros CF. 1991, Identification of broccoli and cauliflower germplasms with RAPD markers. Plant Cell Reporter. 10: 505-511
83． Huang H, Layne DR, Kubisiak TL. 2000, RAPD inheritance and diversity in pawpaw (Asimina trilobo). J. Amer. Soc. Hort. Sci., 125(4) : 454-459
84． Kaundun SS, Zhyvoloup A, Park YG. 2000, Evaluation of the genetic polymorphism among elite tea (Camellia sinensis var. sinensis) accessions using RAPD markers. Euphytica, 115: 7-16
85． Kollipara K P, Singh R J, Hymowitz T. 1997, Phylogenetic and genomic relationships in the genus Glycine Willd. Based on sequences from the ITS region of nuclear rDNA. Genome, 40: 57-68
86． Lanner C, Bryngelesson T, Gustafsson M. 1996, Genetic validity of RAPD markers at the intra-and inter-specific level in wild Brassica species with n=9． Theor. Appl. Genet., 98:9-14
87． Lee SH, Choi HS, Kim RH, Lee HY, Nou IS. 1995, Identification of Korean wild tea plants and Japanese green tea germplasms using RAPD markers. J. Kor. Tea Soc., 1(1) : 129-148
88． Lee SH, Nou IS. 1999, Characteristics of chloroplast DNA restriction fragments in Camellia sinensis. J. Kor. Tea Soc., 5(1) : 33-44
89． Maheswaran M, Subudhi PK, Nandi S, Xu JC, Parco A, Yang DC, Huang N. 1997, Polymorphism, distribution, and segregation of AFLP markers in a doubled haploid rice population. Theor. Appl. Genet., 94: 39-45
90． Mark WC, Harold HH. 1991, Silica gel: An ideal material for field preservation of leaf sample for DNA studies. Taxon, 40: 215-220
91． Matsumoto S, Fukui H. 1996, Identification of rose cultivars and clonal plants by random amplified polyphic DNA. Scientia Horticuiturae, 67: 49-54
92． Matsumoto S, Takeuchi A, Hayatsu M, Hondo S. 1994, Molecular cloning of phenylalanine ammonia-lyase cDNA and classification of varieties and germplasms of tea plant (Camellia sinensis) using the tea PAL cDNA probe. Theor. Appl. Genet., 89: 671-675
93． Maughan PJ, Saghai Maroof MA, Buss GR, Huestis GM. 1996, Amplified fragment length polymorphism (AFLP) in soybean: species diversity, inheritance, and near isogenetic line analysis. Theor. Appl. Genet., 93: 392-401
94． Millan T, Osuna F, Cobos A, Torres AM, Cubero JI. 1996, Using RAPDs to study phylogenetic relationships in Rosa. Theor. Appl. Genet., 92: 273-277
95． Miller JC, Ranksley SD. 1990, RFLP analysis of phylogenetic relationships and genetic variation in the genus Lycopersicon. Theor. Appl. Genet., 80: 437-448
96． Mirshra RK, Sen-Mandi S. 2001, Genome analysis and isozyme studies for developing molecular markers associated with drought tolerance in tea plant. In: Proceedings of 2001 International Conference on O-Cha (Tea) Culture and Science (Session II), pp: 66-69, Oct. 5-8,2001 Shizuoka, Japan
97． Miyashita NT, Kawabe A, Innan H. 1999, DNA variation in the wild plant Arabidopsis thaliana revealed by amplified fragment length polymorphism analysis. Genetics, 152: 1723-1731
98． Nei M, Li WH. 1979, Mathematical model for studying genetic variation in terms of restriction endonuclease. Proc. Natl. Acad. Sci. USA, 76(10) : 5269-5273
99． Olmstea RG, Palmer JD. 1994, Chloroplast DNA systematics: A review of method and data analysis. Amer. J. Bot., 81: 1205-1224
100． Oraguize NC, Gardiner SE, Basset HCM, Stefanati M, Ball RD, Bus VGM, White AG. 2001, Genetic polymorphism and relationship in Malus sp. germplasm collections as determined by randomly amplified polymorphic DNA. J. Amer. Soc. Hon. Sci., 126(3) : 318-328
101． Park YG, Shiv SK. 2000, Genetic variation on wild tea populations in Korea. In Proceedings of the International Symposium on Molecular Biology and Tea
Breeding (Liang YR, Liu ZS, Park YG, Takeda Y, Tanaka J, Lu JL and Zhao D eds), Hangzhou, China November 20-28, 2000． pp: 1-18
102． Parfitt DE, Badenes ML. 1997, Phylogeny of the genus Pistacia determined from analysis of chloroplast genome. Proc. Natl. Acad. Sci. USA, 94: 7987-7992
103． Paul S, Wachira FN, Powell W, Waugh R. 1997, Polymorphism and genetic differentiation among populations of Indian and Kenyan tea [Camellia sinensis (L.) O. Kuntze] revealed by AFLP markers. Theor. Appl. Genet., 94: 255-263
104． Penner GA. 1996, RAPD analysis of plant genomes. In: Jauhar PP eds. Methods of genomes analysis of plants. CRC press, New York, pp: 251-268
105． Persson HA, Rumpunen K Mollerstedt KL. 2000, Identification of culinary rhubarb (Rheum spp.) cultivars using morphological characteristics and RAPD markers. Journal of Horticultural Science and Biotechnology, 75(6) : 684-689
106． Pillay M, Hilu KW. 1995, Chloroplast DNA restriction site analysis in the genus Bromus (Poaceae). Amer. J. Bot., 83: 239-250
107． Ravishankar KV, Anand L, Dinesh MR. 2000, Assessment of genetic relatedness among mango cultivars of India using RAPD markers. Journal of Horticultural Science and Biotechnology, 75(2) : 198-201
108． Sealy JR. 1958, A revision of the genus Camellia. The Royal Horticultural Society, London, pp: 111-131
109． Sedra, MH, Lasherms P, Trouslot P, Combes M, Hamon S. 1998 Identification and genetic diversity analysis of date palm (Phoenix dactylifera L.) varieties from Morocco using RAPD markers. Euphytica, 103: 75-82
110． Struss D, Boritzki M, Glozer K, Southwick M. 2001, Detection of genetic diversity among population of sweet cherry (Prunus avium L.) by AFLPs. Journal of Horticultural Science and Biotechnology, 76(3) : 362-367
111． Suh. Y, Thien LB, Reeve HE. 1993, Molecular evolution and phylogenetic implications of internal transcribed spacer sequences of ribosomal DNA in Winteraceae. Amer. J. Bot., 80: 1042-1055
112． Tanaka J, Yamaguchi N, Nakamura Y. 2001, Pollen parent of tea cultivar
Sayamakaori with insect and cold resistance may not exist. Breed. Res., 3: 43-48
113． Vos P, Rogers R, Bleeker M, Reijans M, Lee TVD, Homes M, Frijters JP, Peleman J, Kuiper M. 1995, AFLP: a new technique of DNA fingerprinting. Nucl. Acids Res., 23: 4407-4414
114． Wachira FN, Tanaka J. Takeda Y. 2001, Genetic variation and differentiation in tea (Camellia sinensis) germplasm revealed by RAPD and AFLP variation. Journal of Horticultural Science and Biotechnology, 76(5) : 557-563
115． Wachira FN, Waugh R, Hackett CA, Powell W. 1995, Detection of genetic diversity in tea (Camellia sinensis) using RAPD markers. Genome. 38: 201-210
116． Wachira FN. 1996, Genetic polymorphism in tea revealed by randomly amplified polymorphic DNA markers. Tea, 17(2) : 60-68
117． Wachira FN. 1997a, Characterization and estimation of genetic relatedness among heterogeneous populations of commercial tea closes by random amplification of genomic DNA samples. Tea, 18(1) : 11-20
118． Wachira FN, Powell W, Waugh R. 1997b, An assessment of genetic diversity among Camellia sinensis L. (cultivated tea) and its wild relatives based on randomly amplified polymorphic DNA and organelle-specific STS. Heredity, 78: 603-611
119． Wang ZY, Second G, Tanksley SD. 1992, Polymorphism and phylogenetic relationship among species in the genus Oryza as determined by analysis of nuclear RFLPs. Theor. Appl. Genet., 83: 565-581．
120． Wendel JF, Schnabel AS, Seelanan T. 1995, Bidirectional interlocus concerted evolution following allopolyploid speciation in cotton (Gossypium). Proc. Natl. Acad. Sci. USA, 92: 280-284
121． White TJ, Brims T, Lee S, Taylor J. 1990． Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand D, Sninsky J, White T eds. PCR Protocols: A Guide to Methods and Application. San Diego, California: Academic Press, pp: 315-322
122. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990, DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl. Acids Res., 18(22): 6531-6535
123. Yang X, Quiros C. 1993, Identification and classification of celery cultivars with RAPD markers. Theor. Appl. Genet., 86:205-212
124. Yoon MS, Doi K, Kaga A, Tomooka N, Vaughan DA. 2000, Analysis of the genetic diversity in the Vigna minima complex and related species in Esta Asia. J. Plant Res., 113:375-386
125. Yuan YM, Kupfer P, Doyle JJ. 1996, Infrageneric phylogeny of the genus Gentiana inferred from nucleotide sequences of the internal transcribed spacers of the nuclear ribosomal DNA. Amer. J. Bot., 83:641-652
126. Zabeau M, Vos P. 1993, Selective restriction fragment amplification: A general method for DNA fingerprinting. European Patent Application No. 92402629.7
127. Zhang DL, Dirr MA, Price RA. 2000, Discrimination and genetic diversity of Cephalotaxus accessions using AFLP markers. J. Amer. Soc. Hort. Sci., 125(4): 404-412
128．松元哲，竹内敦子，山口聪．1997，RFLP解析韩国分类评価．茶业研究报告，85(别册)：20-21
129．松元哲，竹内敦子，山口聪，朴竜求．1998，韩国RFLP解析．茶业研究报告，87(别册)：38-39
130．松元哲，竹内敦子，近藤贞昭．1994，绿茶品种(PAL)遗伝的多样性．茶业研究报告，79(别册)：36-38
131．松元哲，竹内敦子．1996，中国种(PAL)cDNA用RFLP解析．茶业研究报告，84(别册)：42-43
2．王文，凌发瑶，施立明．1994，银额果蝇自然群体中的mtDNA多态性研究(Ⅱ)．银额果蝇的起源和分化．遗传学报，21(4)：263-274
3．王建波，张文驹，陈家宽．1999，核rDNA的ITS序列在被子植物系统与进化研究中的应用．植物分类学报，37(4)：407-416
4．王斌，李松涛，1995，RAPD及其在植物分子生物学研究中的应用．见匡廷云，荆玉祥，李德葆主编．植物分子生物学研究——成就与前景．北京：科学出版社，pp：295-303
5．冯丽春，杨光伟，余茂德、张孝勇、向仲怀．1997，利用RAPD对桑属植物种间亲缘关系的研究．中国农业科学，30(1)：52-56
6．叶创兴．1987，山茶属三新种．中山大学学报(自然科学版)，(1)：17-20
7．刘春林，官春云，李恂．2000，关于植物随机引物扩增多态性DNA标记的可靠性问题．植物生理学通讯，36(1)：56-59
8．向巧萍，向秋云，Liston A，傅立国，傅德志．2000，ITS(nrDNA)片段在冷杉属植物中的长度多态性及其在松科的系统与演化中的应用．植物学报，42(9)：946-951
9．庄晚芳，刘祖生，陈文怀．1981，论茶树变种分类．浙江农业大学学报，7(1)：41-47
10．朱旗，任春梅，洪亚辉，罗军武．1994，茶树叶片DNA提取、纯化与检测．湖南农学院学报，20(2)：114-117
11．吴世安，吕海亮，杨继，饶广远，尤瑞麟，葛颂，钟扬．2000，叶绿体DNA片段的RFLP分析在黄精族系统学研究中的应用．植物分类学报，38(2)：97-110
12．吴美贞．1994，Studies on genetic relationship among the Korean native tea trees and physicochemical properties of its green tea.(韩国)高丽大学校大学院农学科(博士学位论文)，PP：27-29
13．张军丽，王铮峰，李鸣光，王伯荪．2000，植物类群研究中的分子标记及其应用．应用生态学报，11(4)：631-636
14．张宏达．1981a，山茶属植物的系统研究．中山大学学报(自然科学版)论丛，(1)：1-124
15．张宏达．1981b，茶树的系统分类．中山大学学报(自然科学版)，(1)：87-99
16．张宏达．1984，茶叶植物资源的订正．中山大学学报(自然科学版)，(1)：1-12
17．张宏达．1990，中国山茶科植物新种．中山大学学报(自然科学版)，29(2)：85-93
18．张芳赐．1980，云南山茶属的二新种．云南植物研究，2(3)：341-344
19．张芳赐，丁渭然，黄毅，陈兴琰，陈国本，唐明德．1990，云南山茶属的三新种．云南植物研究，12(1)：31-34
20．张芳赐．1994，云南山茶属新变种—邦威茶．见《中国古茶树》，中华茶人联谊会，中国茶叶学会编．上海：上海文化出版社，PP：33-34
21．李光涛，梁涛．1990，中国山茶属4种2变种核型研究．广西植物，10(3)：189-197
22．杜琪珍，李名君，刘维华，王海思．1990，茶组植物的化学分类及数值分类．茶叶科学，10(2)：1-12
23．汪小全，邵喻苹，张大明，洪德元．1996a，银杉遗传多样性的RAPD分析．中国科学(C辑)，(26)：436-441
24．汪小全，邵喻苹，张大明，张志宪，洪德元．1996b，RAPD用于遗传多样性和系统学研究中的问题．植物学报，38(12)：954-962
25．汪小全，洪德元．1997，植物分子系统学近五年的研究进展概况．植物分类学报，35(5)：465-480
26．汪劲武．1984，种子植物分类学．高等教育出版社，PP：1-24
27．邱丽娟，Nelson RL,Vodkin LO.1997，利用RAPD标记鉴定大豆种质．作物学报，23(3)：408-417
28．邹喻苹，汪小全，雷一丁，裴颜龙，张志宪．1994，几种濒危植物及其近缘类群总DNA的提取与鉴定．植物学报，36(7)：528-533
29．邹喻苹，葛颂，王晓东．系统与进化植物学中成分子标记．2001，北京：科学出版社
30．闵天禄．1992，山茶属茶组植物的订正．云南植物研究，14(2)：115-132
31．闵天禄主编．2000，世界山茶属的研究．昆明：云南科技出版社，PP：1-313
32．陈大明，张上隆，金勇丰．1997，一种木本果树基因组DNA提取方法研究．浙江农业大学学报，23(6)：621-624
33．陈亮，杨亚军．1999，茶树分子标记和基因克隆研究进展．茶叶文摘，13(5)：1-6
34．陈亮，杨亚军．2000，RAPD及其在茶树种质资源和遗传育种研究中的应用．海峡两岸茶叶科技学术研讨会．福州4／26-29pp：54-58
35．陈亮，童启庆，高其康，束际林，虞富莲．1997，山茶属8种1变种花粉形态比较．茶叶科学，17(2)：183-188
36．陈亮，虞富莲，童启庆．2000，关于茶组植物分类与演化的讨论．茶叶科学，20(2)：89-94
37．陈亮．1996，茶组植物系统分类学研究现状．茶叶，22(2)：16-19
38．周奕华，陈正华．1999，分子标记在植物学中的应用及前景．武汉植物学研究，17(1)：75-86
39．金惠淑，梁月荣，陆建良．2001，中、韩两国主要茶树品种DNA多态性比较研究．茶叶科学，21(2)：103-107
40．施苏华，唐绍清，陈月琴，屈良鹄，张宏达．1998，11种金花茶植物的RAPD分析及其系统学意义．植物分类学报，36(4)：465-480
41．贾建航，王萍，金德敏，曲雪萍，王倩，李传友，翁曼丽，王斌．2000，RAPD标记在紫菜遗传多样性检测和种质鉴定中的应用．植物学报，42(4)：403-407
42．贾继增．1996，分子标记种质资源鉴定和分子标记育种．中国农业科学，29(4)：1-10
43．郭旺珍，张天真，潘家驹．1996，我国棉花主栽品种的RAPD指纹图谱分析．农业生物技术学报，4(2)：129-134
44．钱韦，葛颂，洪德元．2000，采用RAPD和ISSR标记探讨中国疣粒野生稻的遗传多样性．植物学报，42(7)：741-750
45．梁月荣，田中淳一，武田善行．2000，应用RAPD分子标记分析“晚绿”品种的杂交亲本．茶叶科学，20(1)：22-26
46．梁红健，刘敏，钟志宇，吴应祥，李文彬．1996，中国部分兰花品种RAPD分析．园艺学报，23(4)：365-370
47．梁国鲁，周才琼，林蒙嘉，陈家玉，刘君素．1994，贵州大树茶的核型变异与进化．植物分类学报，32(4)：308-315
48．梁盛业，钟业聪．1981，中国山茶科一个新种．中山大学学报(自然科学版)，(3)：118-119
49．彭英．1992，蛋白质亚基的分离及其与茶组植物的分类．中国茶叶，14(5)：10-11
50．彭隽敏，孔清，徐乃瑜．1995，云南小麦、西藏半野生小麦和普通小麦叶绿体DNA限制性内切酶图谱的研究．遗传，17(6)：4-6
51．程须珍，Yang CY.2001，利用RAPD标记鉴定绿豆组植物种间的亲缘关系．中国农业科学，34(2)：216-218
52．虞富莲．1986，论茶树原产地和起源中心．茶叶科学，6(1)：1-8
53．裴颜龙，邵喻苹，尹蓁，汪小全，张志宪，洪德元．1995，矮牡丹和紫斑牡丹RAPD分析初报．植物分类学报，33(4)：350-356
54．谭永济，陈炳环，虞富莲，王海思，王平盛．1984，中国云南茶树新种和新变种．茶叶科学，4(1)：19-30
55．黎星辉，施兆鹏，刘春林，罗军武，沈程文，龚志华．2001，云南大叶茶与汝城白毛茶杂交后代的RAPD鉴定．茶叶科学，21(2)：99-102
56．黎星辉，施兆鹏，刘春林，罗军武，沈程文，李家贤，方华春．2002，汝城白毛茶与两广主要“白毛茶”居群亲缘关系的RAPD分析．茶叶科学，22(2)：79-82
57．戴思兰，陈俊愉，高荣孚，马江生，李文彬．1996，DNA提纯方法对9种菊属植物RAPD的影响．园艺学报，23(2)：169-174
58. Ainouche ML, Bayer R. 1997, On the origins of the tetraploid Bromus species (section Brornus, Poaceae): insights from internal transcribed spacer sequences of nuclear ribosomal DNA. Genome, 730-743
59. Backmann JS, Soller M. 1983, Restriction fragment length polymorphism in genetic improvement: methodologies, mapping and cost. Theor. Appl. Genet., 67:
35-43
60． Baldwin BG. 1993, Molecular phylogenetics of Calycadenia (Compositae) based on ITS sequences of nuclear nbosomal DNA: chromosomal and morphological evolution reexamined. Amer J Bot, 80: 222-238
61． Baldwin BG, Sanderson MJ, Porter JM, Wojciechowski MF, Campbell CS, Donoghue MJ. 1995, The ITS region of nuclear ribosomal DNA: A valuable source of evidence on angiosperm phylogeny. Ann. Missouri Bot. Card., 247-277
62． Belaj A, Trujilo I, Rosa R, Rallo L, Gimenez MJ. 2001, Polymorphism and discrimination capacity of randomly amplified polymorphic markers in an olive germplasm bank. J. Amer. Soc. Hort. Sci., 126(1) : 64-71
63． Boonprakob U, Byrne D H, Graham C J, Okie W R, Bechman T, Smith B R. 2001, Genetic relationships among cultivated diploid plums and their progenitors as determinated by RAPD markers. J. Amer. Soc. Hort. Sci., 126(4) : 451-461
64． Borthakur S. Mondal TK, Parveen SS, Guha A, Sen P, Borthakur A, Deka PC. 1998, Isolation of chloroplast DNA from tea, Camellia sp. Indian Journal of Experimental Biology, 36: 1165-1167
65． Cabrita LF, Aksoy U, Hepaksoy S Leitao JM. 2001, Suitability of isozyme, RAPD and AFLP markers to access genetic differences and relatedness among fig (Ficus carica L.) clones. Scientia Horticulture, 87: 261-273
66． Cano RJ, Poniar HN, Pieniazek NJ, Acra A, Poinar GO. 1993, Amplification and sequencing of DNA from a 120-135-million-year-old weevil. Nature, 363(10) : 536-538
67． Chen L, Yu FL, Luo LH, Tong QQ. 2001, Morphological classification and phylogenetic evolution of section Thea in the genus Camellia. In: Proceedings of 2001 International Conference on O-Cha (Tea) Science (Session II), pp: 29-32, Oct. 5-8,2001 Shizuoka, Japan
68． Conner PJ, Wood BW. 2001, Identification of pecan germplasms and their genetic relatedness as determined by randomly amplified polymorphic DNA analysis. J.
Amer. Soc. Hort. Sci., 126(4) : 474-480
69． Cornelius PL, Seyedsadr M, Crossa J. 1992, Using the shifted multiplicative model to search for "separability" in crop cultivar trials. Theor. Appl. Genet., 84: 161-172
70． Dellaporta SL, Wood J, Hicks JB. 1983, A plant DNA mini preparation, Version II. Plant Mol. Biol. Rep., 1:19-21
71． Demeke T, Adams R P, Chibbar R. 1992, Potenital taxonomic use of randon amplified polymorphic DNA (RAPD): A case study in Brassica. Theor. Appl. Genet., 84: 990-994
72． Dettori MT, Palombi MA, 2000, Identification of Feijoa Sellowiana Berg accessions by RAPD markers. Scientia Horticulturae, 86: 279-290
73． Doyle JJ, Doyle JL. 1987, A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 19: 11-15
74． Elder JR, Turner BJ. 1995, Concerted evolution of repetitive DNA sequences in eukaryotes. Quart. Rev. Biol., 70: 297-319
75． Fatokun CA, Danesh D, Young ND, Stewart EL. 1993, Molecular taxonomic relationships in the genus Vigna based on RFLP analysis. Theor. Appl. Genet., 86: 97-104
76． Federici CT, Fang DQ, Scora RW, Roose ML. 1998, Phylogenetic relationships within the genus citrus (Rutaceae) and related genera as revealed by RFLP and RAPD analysis. Theor. Appl. Genet., 96: 812-822
77． Guillemaut P, Drouard LM. 1992, Isolation of plant DNA: A fast, inexpensive, and reliable methods. Plant Mol. Biol Rep., 10: 60-65
78． Gulsen O, Roose ML. 2001, Chlorplast and nuclear genome analysis of the parenage of lemons. J. Amer. Soc. Hon. Sci., 126(2) : 210-215
79． Hackett CA, Wachira FN, Paul S, Powell W, Waugh R. 2000, Construction of a genetic linkage map for Camellia sinensis (tea). Heredity, 85(4) : 346-355
80． Halward T, Stalker T, LaRue E. 1992, Use of single primer DNA amplification in genetic studies of peanut (Arachis hypogaea L.). Plant mol. Biol., 18: 315-325
81． Hsiao C, Chatterton NJ, Asay KH, Jensen KB. 1995, Phylogenetic relationships of the mono species of wheat tribe, Triticeae (Poaceae) based on nuclear rDNA (ITS) sequence. Theor. Appl. Genet., 90: 389-398
82． Hu J, Quiros CF. 1991, Identification of broccoli and cauliflower germplasms with RAPD markers. Plant Cell Reporter. 10: 505-511
83． Huang H, Layne DR, Kubisiak TL. 2000, RAPD inheritance and diversity in pawpaw (Asimina trilobo). J. Amer. Soc. Hort. Sci., 125(4) : 454-459
84． Kaundun SS, Zhyvoloup A, Park YG. 2000, Evaluation of the genetic polymorphism among elite tea (Camellia sinensis var. sinensis) accessions using RAPD markers. Euphytica, 115: 7-16
85． Kollipara K P, Singh R J, Hymowitz T. 1997, Phylogenetic and genomic relationships in the genus Glycine Willd. Based on sequences from the ITS region of nuclear rDNA. Genome, 40: 57-68
86． Lanner C, Bryngelesson T, Gustafsson M. 1996, Genetic validity of RAPD markers at the intra-and inter-specific level in wild Brassica species with n=9． Theor. Appl. Genet., 98:9-14
87． Lee SH, Choi HS, Kim RH, Lee HY, Nou IS. 1995, Identification of Korean wild tea plants and Japanese green tea germplasms using RAPD markers. J. Kor. Tea Soc., 1(1) : 129-148
88． Lee SH, Nou IS. 1999, Characteristics of chloroplast DNA restriction fragments in Camellia sinensis. J. Kor. Tea Soc., 5(1) : 33-44
89． Maheswaran M, Subudhi PK, Nandi S, Xu JC, Parco A, Yang DC, Huang N. 1997, Polymorphism, distribution, and segregation of AFLP markers in a doubled haploid rice population. Theor. Appl. Genet., 94: 39-45
90． Mark WC, Harold HH. 1991, Silica gel: An ideal material for field preservation of leaf sample for DNA studies. Taxon, 40: 215-220
91． Matsumoto S, Fukui H. 1996, Identification of rose cultivars and clonal plants by random amplified polyphic DNA. Scientia Horticuiturae, 67: 49-54
92． Matsumoto S, Takeuchi A, Hayatsu M, Hondo S. 1994, Molecular cloning of phenylalanine ammonia-lyase cDNA and classification of varieties and germplasms of tea plant (Camellia sinensis) using the tea PAL cDNA probe. Theor. Appl. Genet., 89: 671-675
93． Maughan PJ, Saghai Maroof MA, Buss GR, Huestis GM. 1996, Amplified fragment length polymorphism (AFLP) in soybean: species diversity, inheritance, and near isogenetic line analysis. Theor. Appl. Genet., 93: 392-401
94． Millan T, Osuna F, Cobos A, Torres AM, Cubero JI. 1996, Using RAPDs to study phylogenetic relationships in Rosa. Theor. Appl. Genet., 92: 273-277
95． Miller JC, Ranksley SD. 1990, RFLP analysis of phylogenetic relationships and genetic variation in the genus Lycopersicon. Theor. Appl. Genet., 80: 437-448
96． Mirshra RK, Sen-Mandi S. 2001, Genome analysis and isozyme studies for developing molecular markers associated with drought tolerance in tea plant. In: Proceedings of 2001 International Conference on O-Cha (Tea) Culture and Science (Session II), pp: 66-69, Oct. 5-8,2001 Shizuoka, Japan
97． Miyashita NT, Kawabe A, Innan H. 1999, DNA variation in the wild plant Arabidopsis thaliana revealed by amplified fragment length polymorphism analysis. Genetics, 152: 1723-1731
98． Nei M, Li WH. 1979, Mathematical model for studying genetic variation in terms of restriction endonuclease. Proc. Natl. Acad. Sci. USA, 76(10) : 5269-5273
99． Olmstea RG, Palmer JD. 1994, Chloroplast DNA systematics: A review of method and data analysis. Amer. J. Bot., 81: 1205-1224
100． Oraguize NC, Gardiner SE, Basset HCM, Stefanati M, Ball RD, Bus VGM, White AG. 2001, Genetic polymorphism and relationship in Malus sp. germplasm collections as determined by randomly amplified polymorphic DNA. J. Amer. Soc. Hon. Sci., 126(3) : 318-328
101． Park YG, Shiv SK. 2000, Genetic variation on wild tea populations in Korea. In Proceedings of the International Symposium on Molecular Biology and Tea
Breeding (Liang YR, Liu ZS, Park YG, Takeda Y, Tanaka J, Lu JL and Zhao D eds), Hangzhou, China November 20-28, 2000． pp: 1-18
102． Parfitt DE, Badenes ML. 1997, Phylogeny of the genus Pistacia determined from analysis of chloroplast genome. Proc. Natl. Acad. Sci. USA, 94: 7987-7992
103． Paul S, Wachira FN, Powell W, Waugh R. 1997, Polymorphism and genetic differentiation among populations of Indian and Kenyan tea [Camellia sinensis (L.) O. Kuntze] revealed by AFLP markers. Theor. Appl. Genet., 94: 255-263
104． Penner GA. 1996, RAPD analysis of plant genomes. In: Jauhar PP eds. Methods of genomes analysis of plants. CRC press, New York, pp: 251-268
105． Persson HA, Rumpunen K Mollerstedt KL. 2000, Identification of culinary rhubarb (Rheum spp.) cultivars using morphological characteristics and RAPD markers. Journal of Horticultural Science and Biotechnology, 75(6) : 684-689
106． Pillay M, Hilu KW. 1995, Chloroplast DNA restriction site analysis in the genus Bromus (Poaceae). Amer. J. Bot., 83: 239-250
107． Ravishankar KV, Anand L, Dinesh MR. 2000, Assessment of genetic relatedness among mango cultivars of India using RAPD markers. Journal of Horticultural Science and Biotechnology, 75(2) : 198-201
108． Sealy JR. 1958, A revision of the genus Camellia. The Royal Horticultural Society, London, pp: 111-131
109． Sedra, MH, Lasherms P, Trouslot P, Combes M, Hamon S. 1998 Identification and genetic diversity analysis of date palm (Phoenix dactylifera L.) varieties from Morocco using RAPD markers. Euphytica, 103: 75-82
110． Struss D, Boritzki M, Glozer K, Southwick M. 2001, Detection of genetic diversity among population of sweet cherry (Prunus avium L.) by AFLPs. Journal of Horticultural Science and Biotechnology, 76(3) : 362-367
111． Suh. Y, Thien LB, Reeve HE. 1993, Molecular evolution and phylogenetic implications of internal transcribed spacer sequences of ribosomal DNA in Winteraceae. Amer. J. Bot., 80: 1042-1055
112． Tanaka J, Yamaguchi N, Nakamura Y. 2001, Pollen parent of tea cultivar
Sayamakaori with insect and cold resistance may not exist. Breed. Res., 3: 43-48
113． Vos P, Rogers R, Bleeker M, Reijans M, Lee TVD, Homes M, Frijters JP, Peleman J, Kuiper M. 1995, AFLP: a new technique of DNA fingerprinting. Nucl. Acids Res., 23: 4407-4414
114． Wachira FN, Tanaka J. Takeda Y. 2001, Genetic variation and differentiation in tea (Camellia sinensis) germplasm revealed by RAPD and AFLP variation. Journal of Horticultural Science and Biotechnology, 76(5) : 557-563
115． Wachira FN, Waugh R, Hackett CA, Powell W. 1995, Detection of genetic diversity in tea (Camellia sinensis) using RAPD markers. Genome. 38: 201-210
116． Wachira FN. 1996, Genetic polymorphism in tea revealed by randomly amplified polymorphic DNA markers. Tea, 17(2) : 60-68
117． Wachira FN. 1997a, Characterization and estimation of genetic relatedness among heterogeneous populations of commercial tea closes by random amplification of genomic DNA samples. Tea, 18(1) : 11-20
118． Wachira FN, Powell W, Waugh R. 1997b, An assessment of genetic diversity among Camellia sinensis L. (cultivated tea) and its wild relatives based on randomly amplified polymorphic DNA and organelle-specific STS. Heredity, 78: 603-611
119． Wang ZY, Second G, Tanksley SD. 1992, Polymorphism and phylogenetic relationship among species in the genus Oryza as determined by analysis of nuclear RFLPs. Theor. Appl. Genet., 83: 565-581．
120． Wendel JF, Schnabel AS, Seelanan T. 1995, Bidirectional interlocus concerted evolution following allopolyploid speciation in cotton (Gossypium). Proc. Natl. Acad. Sci. USA, 92: 280-284
121． White TJ, Brims T, Lee S, Taylor J. 1990． Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis M, Gelfand D, Sninsky J, White T eds. PCR Protocols: A Guide to Methods and Application. San Diego, California: Academic Press, pp: 315-322
122. Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990, DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucl. Acids Res., 18(22): 6531-6535
123. Yang X, Quiros C. 1993, Identification and classification of celery cultivars with RAPD markers. Theor. Appl. Genet., 86:205-212
124. Yoon MS, Doi K, Kaga A, Tomooka N, Vaughan DA. 2000, Analysis of the genetic diversity in the Vigna minima complex and related species in Esta Asia. J. Plant Res., 113:375-386
125. Yuan YM, Kupfer P, Doyle JJ. 1996, Infrageneric phylogeny of the genus Gentiana inferred from nucleotide sequences of the internal transcribed spacers of the nuclear ribosomal DNA. Amer. J. Bot., 83:641-652
126. Zabeau M, Vos P. 1993, Selective restriction fragment amplification: A general method for DNA fingerprinting. European Patent Application No. 92402629.7
127. Zhang DL, Dirr MA, Price RA. 2000, Discrimination and genetic diversity of Cephalotaxus accessions using AFLP markers. J. Amer. Soc. Hort. Sci., 125(4): 404-412
128．松元哲，竹内敦子，山口聪．1997，RFLP解析韩国分类评価．茶业研究报告，85(别册)：20-21
129．松元哲，竹内敦子，山口聪，朴竜求．1998，韩国RFLP解析．茶业研究报告，87(别册)：38-39
130．松元哲，竹内敦子，近藤贞昭．1994，绿茶品种(PAL)遗伝的多样性．茶业研究报告，79(别册)：36-38
131．松元哲，竹内敦子．1996，中国种(PAL)cDNA用RFLP解析．茶业研究报告，84(别册)：42-43