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| 湿地松SSR分子标记开发及脂用无性系遗传多样性分析 |
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胡珊1, 张文娟2, 龚斌1, 张志红3, 周光1, 李火根2, 杨春霞1*
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(1. 江西省林业科学院 江西乡土树种良种选育与高效利用江西省重点实验室/南昌市特色香料树种生物育种重点实验室,南昌 330013；2. 南京林业大学 林木遗传与生物技术教育部重点实验室/南方现代林业协同创新中心,南京 210037；3. 峡江县林木良种场,吉安 331409 )
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| 摘要: |
| 湿地松是重要的产脂树种。为探究不同来源脂用无性系的遗传多样性差异及群体分化规律，解析其群体遗传结构与育种材料来源地的关联机制，该研究利用转录组数据开发SSR标记，对172份湿地松脂用无性系进行遗传多样性评估和群体结构分析。采用MISA软件从湿地松转录组Unigene序列中挖掘SSR位点，利用Primer 3.0软件设计特异性引物；经聚丙烯酰胺凝胶电泳筛选多态性引物后，选取12对扩增稳定、多态性丰富的SSR引物，对172份湿地松脂用无性系进行进行PCR扩增，开展遗传多样性参数估算、群体遗传结构分析及主成分分析。结果表明：（1）从68 211条unigenes序列中共鉴定出2 077个SSR位点，以三核苷酸和二核苷酸重复类型为主，重复单元重复次数主要集中在5~6次，重复单元类型以六碱基基元最为丰富，且以AGATGG/ATCTCC重复基元出现频率最高，占六碱基重复基元总数的22.39%；筛选出12对多态性优良的SSR引物，其多态性信息含量（PIC）平均值为0.291，属于中度多态性水平，可用于湿地松遗传评价。（2）群体遗传结构分析将供试群体划分为两个亚群（K=2，ΔK=346），其中亚群A包含25个无性系，主要源自峡江林木良种场的高产脂育种材料；亚群B包含其余147个无性系，广泛分布于江西省其他地区；主成分分析结果与Structure划分高度吻合，第一、二主成分分别解释总遗传变异的25.06%和12.68%；遗传多样性比较显示，亚群B的遗传多样性水平（Na=3.617，I=0.549，h=0.320）显著高于亚群A（Na=1.917，I=0.258，h=0.143）。综上，江西省湿地松脂用无性系遗传多样性较丰富，且存在明显的群体遗传结构，其中峡江林木良种场育种材料遗传基础相对狭窄，而其他地区来源的无性系遗传多样性更为丰富。该研究为江西省及周边相似气候区域的湿地松良种选育及核心种质构建提供了重要的理论依据。 |
| 关键词: 湿地松，脂用无性系，SSR分子标记，遗传多样性，遗传结构 |
| DOI：10.11931/guihaia.gxzw202603047 |
| 分类号: |
| 基金项目:国家重点研发计划子课题（2022YFD2200204-4）；江西省林业科学院青年科技人才培养项目（2023522901）；江西省林业科学院基础研究科研项目--领军人才项目（2025522903）。 |
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| Development of SSR molecular markers in Pinus elliottii and genetic diversity analysis of resin-producing clones |
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HU Shan1,ZHANG Wenjuan2,GONG Bin1,ZHANG Zhihong3,ZHOU Guang1,LI Huogen2,YANG Chunxia1*
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（1. Jiangxi Provincial Key Laboratory of Improved Variety Breeding and Efficient Utilization of Native Tree Species,Nanchang Key Laboratory of Biological Breeding of Special Spice Trees Species,Jiangxi Academy of Forestry,Nanchang 330013, China; 2. Key Laboratory of Forest Genetics ＆ Biotechnology of Ministry of Education,Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing 210037,China; 3. Xiajiang County Forest Seed Farm,Ji’an 331409,China )
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| Abstract: |
| Pinus elliottii is an important resin-producing tree species. This study focused on resin-use clones from different origins. It aimed to explore their genetic diversity differences and population differentiation patterns. It also sought to clarify how population genetic structure relates to the geographic sources of breeding materials. To achieve this, researchers developed SSR markers using transcriptome data. They then evaluated the genetic diversity and analyzed the population structure of 172 slash pine resin-use clones. The goal is to support selection, protection, and utilization of high-resin germplasm.SSR loci were mined from unigene sequences using MISA software. Specific primers were designed with Primer3.0. Polymorphic primers were screened by polyacrylamide gel electrophoresis. Twelve SSR primers with stable amplification and high polymorphism were selected. These primers were used to amplify 172 resin-used clones of P. elliottii. Genetic diversity parameters, population structure, and principal component analysis (PCA) were calculated. The results were as follows: (1) A total of 2,077 SSR loci were identified from 68,211 unigene sequences. Trinucleotide and dinucleotide repeats were the main types. Repeat numbers were mostly 5-6 times. Hexanucleotide motifs were the most abundant type. The AGATGG/ATCTCC motif had the highest frequency, accounting for 22.39% of all hexanucleotide repeats. Twelve SSR primers with good polymorphism were selected. Their average polymorphism information content (PIC) was 0.291, indicating moderate polymorphism. These 12 primers are suitable for genetic evaluation of P. elliottii.(2) Population structure analysis divided the tested population into two subpopulations (K=2, ΔK=346). Subpopulation A contained 25 clones, mainly from high-resin breeding materials of Xiajiang Forest Tree Improvement Station. Subpopulation B contained the remaining 147 clones, widely distributed in other regions of Jiangxi Province. PCA results were highly consistent with Structure analysis. The first two principal components explained 25.06% and 12.68% of total genetic variation, respectively. Genetic diversity comparison showed subpopulation B had significantly higher diversity (Na=3.617, I=0.549, h=0.320) than subpopulation A (Na=1.917, I=0.258, h=0.143).In conclusion, resin-used clones of P. elliottii in Jiangxi Province have rich genetic diversity and obvious population structure. Breeding materials from Xiajiang Forest Tree Improvement Station have a relatively narrow genetic basis. Clones from other regions show richer genetic diversity. This study provides an important theoretical basis for the selective breeding of improved slash pine varieties and the construction of core germplasm collections in Jiangxi Province and adjacent regions with similar climatic conditions. |
| Key words: Pinus elliottii, resin-producing clones, SSR molecular markers, genetic diversity, genetic structure |
