甜瓜(Cucumis melo L.,2n=2x=24)是葫蘆科作物中營養(yǎng)豐富且具有極高經(jīng)濟(jì)價值的水果型蔬菜作物,其果實具有豐富多樣的表型性狀和生理特征。甜瓜果實性狀對消費(fèi)者和市場銷售商的選擇有重要影響。各種基因調(diào)控著甜瓜的果實品質(zhì)性狀,而這些基因又被很多不同的因素調(diào)控。快速發(fā)展的高通量重測序技術(shù)已經(jīng)使大量的作物基因組重測序成為可能,同時也促進(jìn)了開發(fā)新的分子標(biāo)記。近年來,用CAPS分子標(biāo)記開展QTL分析已成為一種十分重要的方法。本研究通過建立新的基于SNP-CAPS分子標(biāo)記,構(gòu)建遺傳連鎖圖譜,進(jìn)行QTL分析,對甜瓜果實性狀的遺傳機(jī)制進(jìn)行了研究。本研究選用MR-1作為母本(P₁),M4-7作為父本(P₂),構(gòu)建F_2:3群體。兩個表型差異顯著的親本通過重測序后,分析兩個親本基因組之間的SNP位點(diǎn),并將這些SNP位點(diǎn)轉(zhuǎn)化成CAPS分子標(biāo)記。分別將母本91.54%的基因組測序數(shù)據(jù)和父本90%基因組測序數(shù)據(jù)比對到甜瓜參考基因組上。在雙親基因組之間共開發(fā)出了2,761,801個SNP,并轉(zhuǎn)化成了7,741個CAPS標(biāo)記。用4種限制性...

【文章頁數(shù)】：119 頁

【文章目錄】：
摘要
Abstract
1 Introduction
    1.1 Fruits
    1.2 Melon introduction and its classification
    1.3 Melon botany and its origin
    1.4 Physiological changes occur during ripening in melon
    1.5 Biochemical changes occur during ripening in melon
        1.5.1 Carbohydrate metabolism
        1.5.2 Organic acids
        1.5.3 Cell wall metabolism
    1.6 Molecular markers
        1.6.1 Categories of molecular markers
        1.6.2 Dominant DNA markers
        1.6.3 Co-dominant markers
    1.7 Sequencing technology
        1.7.1 High-throughput sequencing
        1.7.2 Application of High-throughput sequencing
    1.8 Quantitative trait loci (QTL)
        1.8.1 Mapping population
        1.8.2 Linkage map analysis
        1.8.3 Determination of genetic distance
        1.8.4 Statistical methods to detect QTL
    1.9 Various genetic basis studies for QTL detection in melon and other fruits
    1.10 Objectives of current research
    1.11 Technical route of research
    1.12 Annual research plan
2 Materials and methods
    2.1 Field trials and methods
        2.1.1 Experimental material
        2.1.2 Greenhouse test design
        2.1.3 Seed treatment
        2.1.4 Raising of nursery seedlings
        2.1.5 Selfing of F2:3 plant population
        2.1.6 Phenotypic evaluation for fruit traits
    2.2 Molecular experiment reagents and instruments
        2.2.1 Required reagents
        2.2.2 Molecular experiment instruments
        2.2.3 Preparation of required reagents
    2.3 Molecular experiment procedures
        2.3.1 Sequencing library assembling
        2.3.2 Assembling of CAPS markers by High-throughput sequencing information
        2.3.3 CAPS markers designing
        2.3.4 PCR for CAPS markers verification and genotyping
        2.3.5 Restriction endonuclease (RE) digestion reaction
    2.4 Molecular markers data analysis method
        2.4.1 CAPS markers genotyping data analysis procedure and method
        2.4.2 Genetic map assembling and QTL analysis
        2.4.3 Statistical analysis
3 Results and analysis
    3.1 Analysis of SNPs distribution on chromosomes of melon genome
    3.2 Analysis of CAPS polymorphism screening and DNA detection
    3.3 Analysis of linkage map construction through novel CAPSs
    3.4 Analysis of melon fruit traits
        3.4.1 Analysis of fruit weight
        3.4.2 Analysis of fruit firmness
        3.4.3 Analysis of fruit length and width
        3.4.4 Analysis of fruit shape
        3.4.5 Analysis of flesh firmness
        3.4.6 Analysis of Brix
        3.4.7 Analysis of p H
        3.4.8 Analysis of TA (titratable acidity)
    3.5 Anaysis of QTLs related to fruit traits
        3.5.1 Analysis of fruit weight QTLs
        3.5.2 Analysis of fruit firmness QTLs
        3.5.3 Analysis of flesh firmness QTLs
        3.5.4 Analysis of Brix QTLs
        3.5.5 Analysis of fruit length, width and shape QTLs
        3.5.6 Analysis of p H QTLs
        3.5.7 Analysis of TA (titratable acidity) QTLs
4 Discussion
    4.1 Importance of molecular markers
    4.2 Linkage map assembling by CAPS markers
    4.3 Detection of melon fruit traits QTLs
        4.3.1 Fruit weight QTLs
        4.3.2 Fruit length, width and shape QTLs
        4.3.3 Fruit firmness and flesh firmness QTLs
        4.3.4 Fruit Brix QTLs
        4.3.5 Fruit pH and TA QTLs
    4.4 Innovative points
5 Conclusion
    5.1 Fruit traits segregation
    5.2 SNP-CAPS markers assembling
    5.3 Linkage mapping and QTL analysis
Acknowledgement
References
Appendix
Papers published during the time period of Ph.D education



本文編號：3894233