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基于SNP-CAPS标记的甜瓜遗传连锁图谱构建及果实相关性状QTL分析

发布时间:2024-02-03 13:01
  甜瓜(Cucumis melo L.,2n=2x=24)是葫芦科作物中营养丰富且具有极高经济价值的水果型蔬菜作物,其果实具有丰富多样的表型性状和生理特征。甜瓜果实性状对消费者和市场销售商的选择有重要影响。各种基因调控着甜瓜的果实品质性状,而这些基因又被很多不同的因素调控。快速发展的高通量重测序技术已经使大量的作物基因组重测序成为可能,同时也促进了开发新的分子标记。近年来,用CAPS分子标记开展QTL分析已成为一种十分重要的方法。本研究通过建立新的基于SNP-CAPS分子标记,构建遗传连锁图谱,进行QTL分析,对甜瓜果实性状的遗传机制进行了研究。本研究选用MR-1作为母本(P1),M4-7作为父本(P2),构建F2:3群体。两个表型差异显著的亲本通过重测序后,分析两个亲本基因组之间的SNP位点,并将这些SNP位点转化成CAPS分子标记。分别将母本91.54%的基因组测序数据和父本90%基因组测序数据比对到甜瓜参考基因组上。在双亲基因组之间共开发出了2,761,801个SNP,并转化成了7,741个CAPS标记。用4种限制性...

【文章页数】:119 页

【文章目录】:
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



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