基于SSR與cpDNA標(biāo)記的中國(guó)西部梨與歐洲梨的遺傳差異與分化路線的研究
發(fā)布時(shí)間：2020-11-21 19:54
梨(Pyrus.L)屬于薔薇科植物,在中國(guó)從寒冷地區(qū)到熱帶地區(qū),從西部高原到東部沿海均有種植。中國(guó)是東方梨的主要起源中心,且種質(zhì)資源豐富。在梨的傳播過(guò)程中,形成三個(gè)多樣性中心,即中國(guó)中心,中亞中心和近東中心。SSR標(biāo)記在梨中具有高度多態(tài)性和共顯性遺傳,可以用于評(píng)估遺傳多樣性,闡述遺傳關(guān)系與進(jìn)化路線。由于微衛(wèi)星在梨屬甚至整個(gè)薔薇科家族中具有多態(tài)性豐富,可重復(fù)的共顯性遺傳與可轉(zhuǎn)移性,被證實(shí)是可用的遺傳標(biāo)記并且具有等位基因可變特性等優(yōu)異特點(diǎn)。SSR標(biāo)記和葉綠體分子標(biāo)記分別來(lái)自核基因組和葉綠體基因組,它們可以相互補(bǔ)充并且更全面的闡述某一現(xiàn)象。本研究使用131份梨資源(66份西洋梨,33份新疆梨,4份秋子梨,15份白梨,2份褐梨,2份杏葉梨,4份木梨,1份沙梨,吉爾吉斯斯坦的Kai-4,Kai-9及塔吉克斯坦的Pu-0234和Pu-0234),應(yīng)用17對(duì)SSR引物和5對(duì)葉綠體DNA引物用于評(píng)價(jià)中國(guó)西部和歐洲梨種質(zhì)的遺傳多樣性,闡述中國(guó)西部梨種質(zhì)與歐洲梨種質(zhì)的系統(tǒng)發(fā)育關(guān)系及揭示分化路線。所有SSR標(biāo)記均具有多態(tài)性,在131個(gè)種質(zhì)中共獲得377個(gè)等位基因。通過(guò)多樣性統(tǒng)計(jì)發(fā)現(xiàn)中國(guó)梨種質(zhì)的遺傳多樣性高于西洋梨種質(zhì)(中國(guó)西部梨種質(zhì)的Ho和He分別為0.65和0.84,而西洋梨種質(zhì)為0.57和0.73)。通過(guò)聚類分析發(fā)現(xiàn)東方梨種質(zhì)具有遺傳相關(guān)性。PCoA分析顯示除了杏葉梨和木梨-3相近于西方梨種質(zhì),兩個(gè)地理群體之間的種質(zhì)存在巨大的差異。通過(guò)群體結(jié)構(gòu)分析也表明了大多數(shù)東方與西方種質(zhì)資源具有巨大差異,除了乞力阿木特,夏脆,長(zhǎng)把,八角梨,奎克阿木特,油餃團(tuán),杏葉梨及木梨-3與西洋梨親緣關(guān)系較近,這些結(jié)果與PCoA分析一致。通過(guò)Custer和PCoA分析,把所有種質(zhì)分成兩個(gè)種群是最佳選擇,且所有中國(guó)梨種質(zhì)的遺傳關(guān)系均相近。使用5對(duì)cpDNA通用引物對(duì)中國(guó)和歐洲梨種質(zhì)的遺傳多樣性進(jìn)行評(píng)價(jià)。trnL-trnF-1,trnL-trnF-2,trnS-psbC,rbcL和accD-psaI的單倍型數(shù)目分別為三種,七種,四種,三種和十種單倍型。Hd(0.7348)值在accD-psaI高變區(qū)中最大。Vh和Sh的最大值均在accD-psaI中檢測(cè)到(Vh=0.00236,Sh=0.049),而最小值均在rbcL檢測(cè)到(Vh=0.00044,Sh=0.021)。Tajima’s D值的最小值在accD-psaI(-0.14472)中檢測(cè)到而最大值在rbcL(2.82532)檢測(cè)到。多態(tài)性分離數(shù)為10,單倍型多樣性為0.8333,K平均數(shù)為14.000,最大核苷酸多樣性為0.00302。除多態(tài)性分離數(shù)外,東方梨種質(zhì)中所有指標(biāo)的最高值均高于西方梨種質(zhì)。通過(guò)分析高變區(qū)和基因間隔在9個(gè)物種中共檢測(cè)到15個(gè)單倍型。中國(guó)梨種質(zhì)資源檢測(cè)到H-2至H-15單倍型,西方種質(zhì)中檢測(cè)到H-1至H-6單倍型。未知種質(zhì)與西洋梨種質(zhì)中未檢測(cè)到H-7至H-15單倍型,且所有東方種質(zhì)中未發(fā)現(xiàn)H-1單倍型。通過(guò)對(duì)資源單倍型的地理分布進(jìn)行分析,國(guó)外梨種質(zhì)資源具有H-1,H-5和H-6單倍型,新疆梨、白梨、屬于PO群體的Kréd Sobieshi與Menie資源具有H-2單倍型,H-3單倍型主要存在與國(guó)外梨種質(zhì)中,而東方梨種質(zhì)具有的單倍型幾乎在新疆梨中檢測(cè)不到,表明西洋梨與新疆梨種質(zhì)顯示出相近的遺傳關(guān)系,H-2和H-4分別從新疆到波蘭與新疆到保加利亞的途中發(fā)生了基因?qū)?H-3是從吉爾吉斯斯坦到新疆途中發(fā)生了基因?qū)?新疆可能是東西方梨資源進(jìn)化的重要地方點(diǎn),基因滲透可能發(fā)生在此條線路上。
【學(xué)位單位】：中國(guó)農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位年份】：2019
【中圖分類】：S661.2
【文章目錄】：
摘要
ABSTRACT
ABBREVIATIONS
CHAPTER Ⅰ INTRODUCTION
1.1 GENERAL INFORMATION OF PEAR
1.1.1Brief history of Pyrus
1.1.2 Origin and distribution
1.2.GENETIC DIVERSITY AMONG DIFFERENT TRAITS OF PEAR
1.2.1 Tree Characters
1.2.2 Leaf characters
1.2.3 Flowering characters
1.2.4 Fruit characters
1.2.5 Quality characters
1.3 NUTRITIONAL IMPORTANCE
1.3.1 Composition
1.3.2 Nutritional value and health benefits of pear fruit
1.4 BIODIVERSITY AND PLANT GENETIC RESOURCES
1.4.1 Degree of genetic uniformity/variability in Pyrus
1.4.2 Preservation of germplasm
1.4.3 In vivo conservation
1.4.4 In vitro preservation
1.5 SIMPLE SEQUENCE REPEAT（SSR）MARKER
1.5.1 History
1.5.2 Identification and genetic diversity of SSR marker
1.5.3 Application of SSR marker on origin evolution and kinship identification
1.5.4 Application of SSR marker on gene mapping and genetic map construction
1.5.5 Application of SSR marker on fingerprint and molecular identity card construction
1.5.6 Development of SSR molecular markers on the genome of Prunus
1.6 CHLOROPLAST DNA MARKER
1.6.1 History
1.6.2 Characteristics of Chloroplast DNA
1.6.3 Research status of chloroplast DNA on plants
1.6.4 Research status of chloroplast DNA on fruit trees
1.6.5 Research status of chloroplast DNA in pear germplasm resources
1.7 PURPOSE AND SIGNIFICANCE OF THE STUDY
1.8 OBJECTIVES OF THE PROPOSED STUDY
CHAPTER Ⅱ GENETIC DIVERSITY AND ANALYSIS OF PEAR GERMPLASM RESOURCES USING SSR MARKER
2.1 INTRODUCTION
2.2.MATERIALS AND METHODS
2.2.1 Experimental materials
2.2.2 Test agents and instruments
2.2.3 Experimental main instrument
2.2.4 Isolation of pear DNA in the previous research
2.2.5 Pear genomic DNA extraction in existence study
2.2.6 Identification of genomic DNA
2.2.7 Genomic DNA concentration adjustment
2.2.8 SSR primer screening
2.2.9 Characterization of PCR amplification
2.2.10 SSR primer pairings and PCR amplification
2.2.11 Electrophoresis of amplified DNA
2.2.12 Preparation of polyacrylamide gel
2.2.13 Electrophoresis
2.2.14 Silver Dye
2.2.15 Purification of PCR products
2.2.16 Purification product electrophoresis
2.2.17 Data analysis of simple sequence repeat markers
2.3 RESULTS
2.3.1 Genetic diversity assessment by SSR markers
2.3.2 Principal coordinate analysis of131 pear germplasms
2.4 DISCUSSIONS
2.4.1 Characteristics of genetic evaluation by SSR
2.4.2 Genetic relationship of pear accessions inferred by PCoA analysis
CHAPTER Ⅲ PHYLOGENETIC RELATIONSHIP AND POPULATION STRUCTURE ANALYSIS OF PEAR GERMPLASM RESOURCES USING SSR MARKER
3.1 INTRODUCTION
3.2 MATERIALS AND METHODS
3.2.1 Plant materials
3.2.2 Statistical analysis method
3.3 RESULTS
3.3.1 Cluster analysis of131 pear germplasms by SSR markers
3.3.2 Population Structure of131 Pear Germplasms
3.4 DISCUSSIONS
3.4.1 Genetic relationship of pear accessions by the cluster analysis
3.4.2 Genetic structure and genetic diversity of pear accessions
CHAPTER Ⅳ GENETIC DIVERSITY AND DIFFERENTIATION RELATIONSHIP ANALYSIS OF PEAR GERMPLASM RESOURCES USING CHLOROPLAST DNA MARKER
4.1 INTRODUCTION
4.2 MATERIAL METHOD
4.2.1 Plant material
4.2.2 Determination and screening of universal primers for chloroplast DNA for PCR amplification
4.2.3 Chloroplast DNA selected screened markers
4.2.4 Chloroplast DNA fragments amplification and sequencing
4.2.5 Data analysis of chloroplast DNA regions
4.3 RESULTS
4.3.1 Genetic diversity and haplotypes information of chloroplast DNA
4.3.2 Conservation of cpDNA haplotype by one large deletion
4.3.3 Geographical distribution of chloroplast DNA haplotypes
4.3.4 Median-Joining network for chloroplast DNA
4.4 DISCUSSIONS
4.4.1 Genetic diversity and haplotype information of chloroplast DNA
4.4.2 Construction of an intermediate network map between haplotypes
CHAPTER Ⅴ FULL-TEXT CONCLUSION
REFERENCES
ACKNOWLEDGEMENT
CURRICULUM VITAE
【參考文獻(xiàn)】
本文編號(hào)：2893519
【學(xué)位單位】：中國(guó)農(nóng)業(yè)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位年份】：2019
【中圖分類】：S661.2
【文章目錄】：
摘要
ABSTRACT
ABBREVIATIONS
CHAPTER Ⅰ INTRODUCTION
1.1 GENERAL INFORMATION OF PEAR
1.1.1Brief history of Pyrus
1.1.2 Origin and distribution
1.2.GENETIC DIVERSITY AMONG DIFFERENT TRAITS OF PEAR
1.2.1 Tree Characters
1.2.2 Leaf characters
1.2.3 Flowering characters
1.2.4 Fruit characters
1.2.5 Quality characters
1.3 NUTRITIONAL IMPORTANCE
1.3.1 Composition
1.3.2 Nutritional value and health benefits of pear fruit
1.4 BIODIVERSITY AND PLANT GENETIC RESOURCES
1.4.1 Degree of genetic uniformity/variability in Pyrus
1.4.2 Preservation of germplasm
1.4.3 In vivo conservation
1.4.4 In vitro preservation
1.5 SIMPLE SEQUENCE REPEAT（SSR）MARKER
1.5.1 History
1.5.2 Identification and genetic diversity of SSR marker
1.5.3 Application of SSR marker on origin evolution and kinship identification
1.5.4 Application of SSR marker on gene mapping and genetic map construction
1.5.5 Application of SSR marker on fingerprint and molecular identity card construction
1.5.6 Development of SSR molecular markers on the genome of Prunus
1.6 CHLOROPLAST DNA MARKER
1.6.1 History
1.6.2 Characteristics of Chloroplast DNA
1.6.3 Research status of chloroplast DNA on plants
1.6.4 Research status of chloroplast DNA on fruit trees
1.6.5 Research status of chloroplast DNA in pear germplasm resources
1.7 PURPOSE AND SIGNIFICANCE OF THE STUDY
1.8 OBJECTIVES OF THE PROPOSED STUDY
CHAPTER Ⅱ GENETIC DIVERSITY AND ANALYSIS OF PEAR GERMPLASM RESOURCES USING SSR MARKER
2.1 INTRODUCTION
2.2.MATERIALS AND METHODS
2.2.1 Experimental materials
2.2.2 Test agents and instruments
2.2.3 Experimental main instrument
2.2.4 Isolation of pear DNA in the previous research
2.2.5 Pear genomic DNA extraction in existence study
2.2.6 Identification of genomic DNA
2.2.7 Genomic DNA concentration adjustment
2.2.8 SSR primer screening
2.2.9 Characterization of PCR amplification
2.2.10 SSR primer pairings and PCR amplification
2.2.11 Electrophoresis of amplified DNA
2.2.12 Preparation of polyacrylamide gel
2.2.13 Electrophoresis
2.2.14 Silver Dye
2.2.15 Purification of PCR products
2.2.16 Purification product electrophoresis
2.2.17 Data analysis of simple sequence repeat markers
2.3 RESULTS
2.3.1 Genetic diversity assessment by SSR markers
2.3.2 Principal coordinate analysis of131 pear germplasms
2.4 DISCUSSIONS
2.4.1 Characteristics of genetic evaluation by SSR
2.4.2 Genetic relationship of pear accessions inferred by PCoA analysis
CHAPTER Ⅲ PHYLOGENETIC RELATIONSHIP AND POPULATION STRUCTURE ANALYSIS OF PEAR GERMPLASM RESOURCES USING SSR MARKER
3.1 INTRODUCTION
3.2 MATERIALS AND METHODS
3.2.1 Plant materials
3.2.2 Statistical analysis method
3.3 RESULTS
3.3.1 Cluster analysis of131 pear germplasms by SSR markers
3.3.2 Population Structure of131 Pear Germplasms
3.4 DISCUSSIONS
3.4.1 Genetic relationship of pear accessions by the cluster analysis
3.4.2 Genetic structure and genetic diversity of pear accessions
CHAPTER Ⅳ GENETIC DIVERSITY AND DIFFERENTIATION RELATIONSHIP ANALYSIS OF PEAR GERMPLASM RESOURCES USING CHLOROPLAST DNA MARKER
4.1 INTRODUCTION
4.2 MATERIAL METHOD
4.2.1 Plant material
4.2.2 Determination and screening of universal primers for chloroplast DNA for PCR amplification
4.2.3 Chloroplast DNA selected screened markers
4.2.4 Chloroplast DNA fragments amplification and sequencing
4.2.5 Data analysis of chloroplast DNA regions
4.3 RESULTS
4.3.1 Genetic diversity and haplotypes information of chloroplast DNA
4.3.2 Conservation of cpDNA haplotype by one large deletion
4.3.3 Geographical distribution of chloroplast DNA haplotypes
4.3.4 Median-Joining network for chloroplast DNA
4.4 DISCUSSIONS
4.4.1 Genetic diversity and haplotype information of chloroplast DNA
4.4.2 Construction of an intermediate network map between haplotypes
CHAPTER Ⅴ FULL-TEXT CONCLUSION
REFERENCES
ACKNOWLEDGEMENT
CURRICULUM VITAE
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 胡春云;鄭小艷;滕元文;;梨屬葉綠體非編碼區(qū)trnL-trnF和accD-psaI特征及其在系統(tǒng)發(fā)育研究中的應(yīng)用價(jià)值[J];園藝學(xué)報(bào);2011年12期
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