分子标记技术在桃花品种系统分类中的应用研究
摘要
本研究是作者12年来在对国内桃花品种资源调查研究的基础上，结合近三年来从日本农林水产省筑波果树实验场及美国北卡罗来纳州J．C．若斯顿树木园收集的共51个桃花品种，应用AFLP和ISSR分子标记技术，结合形态学特征，进行桃花品种系统分类研究，并建立起桃花品种分类系统及DNA指纹图谱库。
本研究所有实验部分在哈佛大学标本馆分子生物学实验室完成。中英文两个版本的论文写作在美国缅因大学完成。最终定稿于北京林业大学。主要结论如下：
应用AFLP分子标记技术，对51个桃(Prunus persica)与山桃(Prunus davidiana)的品种及变种加以研究。6对AFLP引物组合(EcoRI／MseI：ACC／CAT，AGG／CAT，ACT／CAT，ACC／CTC，AGG／CTC，及ACT／CTC)共产生275条有效谱带，其中93％为多态性标记。AFLP分析结果显示，枝型是相当重要的桃花品种分类特征，应该在桃花品种分类系统中置于显要地位。
应用ISSR分子标记研究来自桃花和山桃的16个品种(变种)间遗传关系。应用10个ISSR引物(UBC818，UBC825，UBC834，UBC855，UBC817，UBC868，UBC845，UBC899，UBC860和UBC836)产生132条谱带，其中62％为多态性谱带。分析结果表明，ISSR技术可以有效地揭示桃花品种间的遗传关系，特别是阐明来自相近种源的桃花品种内部的组别关系。从目前所有可应用的100个ISSR引物中筛选出10个相对于桃花ISSR反应体系最为理想的引物，建立了适合于桃花ISSR实验的反应体系及操作程序，为今后进一步应用这一技术在桃花品种鉴定、品种保护及登录奠定了基础。
AFLP和ISSR两种分子标记技术结合共同应用于桃花品种间遗传关系的研究。这两种方法在桃花枝型分析上的结果极为一致。实验结果表明，无论是AFLP还是ISSR方法，在桃花品种种质鉴别，评价品种间遗传关系等方面极具潜力。此前国内外尚未有关于AFLP和ISSR技术应用于观赏桃花研究的报道。
用6个AFLP引物组合建立了桃花品种DNA指纹图谱库。这些分子数据可以用来鉴别或支持桃花品种的描述及登录，证实桃花品种的育种起源，为今后新品种的保护、鉴定和登录奠定了坚实的基础。
桃花是中国古老的传统名花之一，在我国有3000多年的栽培历史。目前世界上还没有专门的桃花品种登录机构。本研究应用分子标记技术建立的桃花品种分类系统及DNA指纹图谱库为我国申报桃花品种国际登录权威奠定了坚实的基础。这一系统的建立也为今后进一步开展桃花品种鉴定、保护及育种提供了重要的依据。
Based on 12 years studies on ornamental peach (Prunus persica (L.) Batsch.), especially these three years collecting and studying ornamental peach cultivars from Japan and USA, AFLP and ISSR markers had been successfully constructed among 51 ornamental peach taxa. An updated ornamental peach systematics and DNA fingerprinting library had been established with both morphological and molecular evidence.
All 51 ornamental peach taxa were from Beijing Botanical Garden (China), National Institute of Fruit Tree Science, Tsukuba (Japan), and JC Raulston Arboretum at the North Carolina State University, Raleigh, North Carolina (USA) and collected by author herself.
This dissertation is completed in collaboration with Beijing Forestry University, University of Maine and Harvard University. The experiments and datum analyses were completed in both Harvard University and University of Maine. Dissertation (both English and Chinese versions) was written at the University of Maine and revised in Beijing Forestry University.
Both AFLP and ISSR could generate large amount detailed fragments with high polymorphisms and have higher reproducibility compared with other molecular markers. Therefore, these two molecular techniques were applied for this ornamental peach study. No literature on application of AFLP and ISSR for ornamental peach has been found and this innovational work brought their applications further to systematics, genetic relationship, cultivar identification, guided breeding, and nomenclature.
Ornamental peach originates from China. There is 3000 years cultivation history in China. No appointed institution is serving as the International Registration Authority (IRA) for ornamental peach so far in the world. The systematics and DNA fingerprinting library established through this study have provided the supporting justification for China to apply IRA for ornamental peach.
The major results were as follows:
AFLP markers had successfully constructed genetic relationship among 51 ornamental peach taxa. The results showed that AFLP is a powerful technique for detecting the origination and development of ornamental peach taxa.
ISSR markers had successfully constructed genetic relationships among 16 ornamental peach taxa. The results demonstrated that ISSR fingerprinting is an efficient technique to reveal group genetic relationships of ornamental peach. The protocol and ISSR-PCR conditions for ornamental peach had been established.
DNA fingerprinting library of 51 ornamental peach taxa had been established by using 6 AFLP primer combinations. This DNA library is a great example to apply molecular data for the aid of guided breeding and nomenclature for cultivated plants. Molecular data are powerful addition for nomenclature and cultivar registration and should be included in the International Code of Nomenclature for Cultivated Plants (ICNCP). Recommendation provisions based on ornamental peach AFLP analysis were proposed for amending the revision of future ICNCP.
Combined AFLP and ISSR markers provided better genetic information and relationships of 16 ornamental peach taxa. The results suggested that both AFLP and ISSR analysis should have great potential to estimate phylogeny and elucidate the hierarchy among ornamental peach cultivars.
An updated ornamental peach systematics had been established with both morphological and molecular evidence. The nomenclatural status of all known ornamental peach in the world had been reinstated or corrected. A worldwide ornamental peach checklist was presented to apply International Registration Authority for ornamental peach by Beijing Botanical Garden in Beijing, China.
本研究所有实验部分在哈佛大学标本馆分子生物学实验室完成。中英文两个版本的论文写作在美国缅因大学完成。最终定稿于北京林业大学。主要结论如下：
应用AFLP分子标记技术，对51个桃(Prunus persica)与山桃(Prunus davidiana)的品种及变种加以研究。6对AFLP引物组合(EcoRI／MseI：ACC／CAT，AGG／CAT，ACT／CAT，ACC／CTC，AGG／CTC，及ACT／CTC)共产生275条有效谱带，其中93％为多态性标记。AFLP分析结果显示，枝型是相当重要的桃花品种分类特征，应该在桃花品种分类系统中置于显要地位。
应用ISSR分子标记研究来自桃花和山桃的16个品种(变种)间遗传关系。应用10个ISSR引物(UBC818，UBC825，UBC834，UBC855，UBC817，UBC868，UBC845，UBC899，UBC860和UBC836)产生132条谱带，其中62％为多态性谱带。分析结果表明，ISSR技术可以有效地揭示桃花品种间的遗传关系，特别是阐明来自相近种源的桃花品种内部的组别关系。从目前所有可应用的100个ISSR引物中筛选出10个相对于桃花ISSR反应体系最为理想的引物，建立了适合于桃花ISSR实验的反应体系及操作程序，为今后进一步应用这一技术在桃花品种鉴定、品种保护及登录奠定了基础。
AFLP和ISSR两种分子标记技术结合共同应用于桃花品种间遗传关系的研究。这两种方法在桃花枝型分析上的结果极为一致。实验结果表明，无论是AFLP还是ISSR方法，在桃花品种种质鉴别，评价品种间遗传关系等方面极具潜力。此前国内外尚未有关于AFLP和ISSR技术应用于观赏桃花研究的报道。
用6个AFLP引物组合建立了桃花品种DNA指纹图谱库。这些分子数据可以用来鉴别或支持桃花品种的描述及登录，证实桃花品种的育种起源，为今后新品种的保护、鉴定和登录奠定了坚实的基础。
桃花是中国古老的传统名花之一，在我国有3000多年的栽培历史。目前世界上还没有专门的桃花品种登录机构。本研究应用分子标记技术建立的桃花品种分类系统及DNA指纹图谱库为我国申报桃花品种国际登录权威奠定了坚实的基础。这一系统的建立也为今后进一步开展桃花品种鉴定、保护及育种提供了重要的依据。
Based on 12 years studies on ornamental peach (Prunus persica (L.) Batsch.), especially these three years collecting and studying ornamental peach cultivars from Japan and USA, AFLP and ISSR markers had been successfully constructed among 51 ornamental peach taxa. An updated ornamental peach systematics and DNA fingerprinting library had been established with both morphological and molecular evidence.
All 51 ornamental peach taxa were from Beijing Botanical Garden (China), National Institute of Fruit Tree Science, Tsukuba (Japan), and JC Raulston Arboretum at the North Carolina State University, Raleigh, North Carolina (USA) and collected by author herself.
This dissertation is completed in collaboration with Beijing Forestry University, University of Maine and Harvard University. The experiments and datum analyses were completed in both Harvard University and University of Maine. Dissertation (both English and Chinese versions) was written at the University of Maine and revised in Beijing Forestry University.
Both AFLP and ISSR could generate large amount detailed fragments with high polymorphisms and have higher reproducibility compared with other molecular markers. Therefore, these two molecular techniques were applied for this ornamental peach study. No literature on application of AFLP and ISSR for ornamental peach has been found and this innovational work brought their applications further to systematics, genetic relationship, cultivar identification, guided breeding, and nomenclature.
Ornamental peach originates from China. There is 3000 years cultivation history in China. No appointed institution is serving as the International Registration Authority (IRA) for ornamental peach so far in the world. The systematics and DNA fingerprinting library established through this study have provided the supporting justification for China to apply IRA for ornamental peach.
The major results were as follows:
AFLP markers had successfully constructed genetic relationship among 51 ornamental peach taxa. The results showed that AFLP is a powerful technique for detecting the origination and development of ornamental peach taxa.
ISSR markers had successfully constructed genetic relationships among 16 ornamental peach taxa. The results demonstrated that ISSR fingerprinting is an efficient technique to reveal group genetic relationships of ornamental peach. The protocol and ISSR-PCR conditions for ornamental peach had been established.
DNA fingerprinting library of 51 ornamental peach taxa had been established by using 6 AFLP primer combinations. This DNA library is a great example to apply molecular data for the aid of guided breeding and nomenclature for cultivated plants. Molecular data are powerful addition for nomenclature and cultivar registration and should be included in the International Code of Nomenclature for Cultivated Plants (ICNCP). Recommendation provisions based on ornamental peach AFLP analysis were proposed for amending the revision of future ICNCP.
Combined AFLP and ISSR markers provided better genetic information and relationships of 16 ornamental peach taxa. The results suggested that both AFLP and ISSR analysis should have great potential to estimate phylogeny and elucidate the hierarchy among ornamental peach cultivars.
An updated ornamental peach systematics had been established with both morphological and molecular evidence. The nomenclatural status of all known ornamental peach in the world had been reinstated or corrected. A worldwide ornamental peach checklist was presented to apply International Registration Authority for ornamental peach by Beijing Botanical Garden in Beijing, China.
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