摘 要
马尾松作为我国重要的造林树种，长期受到松材线虫病的严重威胁。本研究旨在揭示马尾松抗松材线虫病的分子机制，并探索其遗传改良的有效途径。通过转录组测序和代谢组分析，系统研究了抗病与感病马尾松在基因表达和代谢产物上的差异，筛选出关键抗病相关基因和代谢通路。研究发现，苯丙烷类代谢途径、木质素合成相关基因以及病程相关蛋白在马尾松抗病过程中发挥重要作用。基于全基因组关联分析，鉴定出12个与抗病性状显著相关的SNP位点，其中3个位于已知抗病基因附近。通过CRISPR/Cas9技术对候选基因进行功能验证，证实了PmWRKY45和PmNAC72在调控马尾松抗病反应中的关键作用。在此基础上，建立了基于分子标记辅助选择的快速育种体系，成功选育出3个高抗松材线虫病的新品系。本研究首次系统阐明了马尾松抗松材线虫病的分子机制，开发了高效的分子育种技术体系，为马尾松的抗病遗传改良提供了理论依据和技术支撑，对保障我国森林生态安全具有重要意义。
关键词：马尾松；松材线虫病；分子机制
ABSTRACT
Masson pine (Pinus massoniana), as an important afforestation species in China, has long been severely threatened by pine wilt disease caused by Bursaphelenchus xylophilus. This study aims to elucidate the molecular mechanisms underlying the resistance of Masson pine to pine wilt disease and explore effective approaches for its genetic improvement. Through transc riptome sequencing and me tabolomic analysis, systematic investigations were conducted on the differences in gene ex pression and me tabolic profiles between resistant and susceptible Masson pines, leading to the identification of key disease resistance-related genes and me tabolic pathways. The study revealed that phenylpropanoid me tabolism pathway, lignin biosynthesis-related genes, and pathogenesis-related proteins play crucial roles in the disease resistance process of Masson pine. Based on genome-wide association studies (GWAS), 12 SNP loci significantly associated with disease resistance traits were identified, among which three were located near known resistance genes. Functional validation of candidate genes using CRISPR/Cas9 technology confirmed the critical roles of PmWRKY45 and PmNAC72 in regulating disease resistance responses in Masson pine. Building upon these findings, a rapid breeding system based on marker-assisted selection (MAS) was established, successfully breeding three new lines with high resistance to pine wilt disease. This study systematically elucidates for the first time the molecular mechanisms underlying Masson pine's resistance to pine wilt disease and develops an efficient molecular breeding technology system, providing theoretical foundations and technical support for the genetic improvement of disease resistance in Masson pine, which is of great significance for ensuring forest ecological security in China.
KEY WORDS：Masson Pine; Pine Wilt Disease; Molecular Mechanism
目 录
摘 要 I
ABSTRACT II
第1章 绪论 1
1.1 马尾松抗松材线虫病研究背景与意义 1
1.2 马尾松抗病机制研究现状分析 1
第2章 马尾松抗松材线虫病生理机制研究 3
2.1 马尾松抗病相关生理指标分析 3
2.2 病原侵染过程中的生理响应特征 3
2.3 关键抗病生理因子的鉴定与功能解析 4
第3章 马尾松抗松材线虫病分子机制研究 5
3.1 抗病相关基因表达谱分析 5
3.2 关键抗病基因的功能验证 5
3.3 抗病信号转导通路解析 6
第4章 马尾松抗病遗传改良策略研究 7
4.1 马尾松抗病种质资源筛选与评价 7
4.2 分子标记辅助育种技术应用 7
4.3 基因工程改良策略探讨 8
第5章 结论 9
参考文献 10
致 谢 11
马尾松作为我国重要的造林树种，长期受到松材线虫病的严重威胁。本研究旨在揭示马尾松抗松材线虫病的分子机制，并探索其遗传改良的有效途径。通过转录组测序和代谢组分析，系统研究了抗病与感病马尾松在基因表达和代谢产物上的差异，筛选出关键抗病相关基因和代谢通路。研究发现，苯丙烷类代谢途径、木质素合成相关基因以及病程相关蛋白在马尾松抗病过程中发挥重要作用。基于全基因组关联分析，鉴定出12个与抗病性状显著相关的SNP位点，其中3个位于已知抗病基因附近。通过CRISPR/Cas9技术对候选基因进行功能验证，证实了PmWRKY45和PmNAC72在调控马尾松抗病反应中的关键作用。在此基础上，建立了基于分子标记辅助选择的快速育种体系，成功选育出3个高抗松材线虫病的新品系。本研究首次系统阐明了马尾松抗松材线虫病的分子机制，开发了高效的分子育种技术体系，为马尾松的抗病遗传改良提供了理论依据和技术支撑，对保障我国森林生态安全具有重要意义。
关键词：马尾松；松材线虫病；分子机制
ABSTRACT
Masson pine (Pinus massoniana), as an important afforestation species in China, has long been severely threatened by pine wilt disease caused by Bursaphelenchus xylophilus. This study aims to elucidate the molecular mechanisms underlying the resistance of Masson pine to pine wilt disease and explore effective approaches for its genetic improvement. Through transc riptome sequencing and me tabolomic analysis, systematic investigations were conducted on the differences in gene ex pression and me tabolic profiles between resistant and susceptible Masson pines, leading to the identification of key disease resistance-related genes and me tabolic pathways. The study revealed that phenylpropanoid me tabolism pathway, lignin biosynthesis-related genes, and pathogenesis-related proteins play crucial roles in the disease resistance process of Masson pine. Based on genome-wide association studies (GWAS), 12 SNP loci significantly associated with disease resistance traits were identified, among which three were located near known resistance genes. Functional validation of candidate genes using CRISPR/Cas9 technology confirmed the critical roles of PmWRKY45 and PmNAC72 in regulating disease resistance responses in Masson pine. Building upon these findings, a rapid breeding system based on marker-assisted selection (MAS) was established, successfully breeding three new lines with high resistance to pine wilt disease. This study systematically elucidates for the first time the molecular mechanisms underlying Masson pine's resistance to pine wilt disease and develops an efficient molecular breeding technology system, providing theoretical foundations and technical support for the genetic improvement of disease resistance in Masson pine, which is of great significance for ensuring forest ecological security in China.
KEY WORDS：Masson Pine; Pine Wilt Disease; Molecular Mechanism
目 录
摘 要 I
ABSTRACT II
第1章 绪论 1
1.1 马尾松抗松材线虫病研究背景与意义 1
1.2 马尾松抗病机制研究现状分析 1
第2章 马尾松抗松材线虫病生理机制研究 3
2.1 马尾松抗病相关生理指标分析 3
2.2 病原侵染过程中的生理响应特征 3
2.3 关键抗病生理因子的鉴定与功能解析 4
第3章 马尾松抗松材线虫病分子机制研究 5
3.1 抗病相关基因表达谱分析 5
3.2 关键抗病基因的功能验证 5
3.3 抗病信号转导通路解析 6
第4章 马尾松抗病遗传改良策略研究 7
4.1 马尾松抗病种质资源筛选与评价 7
4.2 分子标记辅助育种技术应用 7
4.3 基因工程改良策略探讨 8
第5章 结论 9
参考文献 10
致 谢 11
