大豆是重要的粮油兼用作物，也是人类食用植物油脂的重要来源。大豆油脂是重要性状，挖掘相关基因，开发分子标记，推动生物育种进程具有重要意义。本研究利用齐黄34×东生16分离群体和341份来自各地的大豆种质材料构建的自然群体进行种子含油量表型鉴定，结合连锁分析和全基因组关联分析，定位与大豆含油量相关的关键数量性状（QTL）位点和单核苷酸多态性（SNP）位点，并综合分析两种方法所得结果，挖掘与含油量相关的基因，并利用标记信息对黑龙江省大豆新品种进行单倍型分析。

主要研究结果如下：

（1）利用高油品种齐黄34和东生16杂交衍生的329份重组自交系群体在两年两点下的含油量数据，结合实验室前期利用简化重测序开发得到的6297个SLAF标记构建的遗传图谱进行连锁分析。定位到3个表型贡献率较高的含油量QTL（qOC24-3-1、qOC23-20-1和qOC23-20-2）。其中qOC23-20-2周边还没有已报道过的大豆含油量相关基因位点。统计两年两点的重组自交系群体平均含油量，结合基因型分型数据，对3个含油量QTL间的遗传互作效应展开分析，结果表明qOC23-20-2位点是3个位点中的主效位点，在qOC23-20-2位点范围内筛选到3个候选基因（Glyma.20G237600，Glyma.20G238500和Glyma.20G238800），其中Glyma.20G238500预测具有调控脂质代谢功能，是与油脂合成相关的候选基因。

（2）利用341份材料构成的自然群体两年间的含油量表型数据，结合重测序获得的全基因组范围内1,048,576个SNP标记进行全基因组关联分析，共检测到119个显著性SNP位点，其中第8号染色体关联到多个在两年中与大豆种子含油量有显著相关性的SNP位点。对8号染色体进行候选基因筛选，得到4个候选基因（Glyma.08G110000，Glyma.08G117400，Glyma.08G117600及Glyma.08G123500）。

（3）利用候选基因信息对黑龙江省大豆新品种进行单倍型分析，并开发相关KASP标记，结果表明Glyma.08G117600和Glyma.08G123500对品种含油量的提高效果显著，且对应位点为提高含油量的基因型的品种在黑龙江第一积温带品种中分布较少。因此，Glyma.08G117600及Glyma.08G123500在今后黑龙江省选育高含油量大豆新品种中的运用至关重要。

综上所述，本研究定位到3个高效数量性状位点和119个显著单核苷酸多态性位点，挖掘出7个候选基因，其中Glyma.20G238500，Glyma.08G117600及Glyma.08G123500为大豆高油品种选育应用中的重要候选基因。该研究成果为大豆油脂性状的分子标记辅助选择提供一定的理论依据和技术支持。

Soybean is an important dual-purpose crop for both food and oil production, serving as a vital source of plant oil for human consumption. Soybean oil content represents a crucial trait, making it significant to mine related genes and develop molecular markers, thereby promoting progress in biological breeding. This study utilized a segregating population derived from the cross Qihuang 34 × Dongsheng 16 and a natural population consisting of 341 soybean accessions collected from various regions for phenotypic evaluation of seed oil content. By integrating linkage analysis and genome-wide association study (GWAS), key quantitative trait loci (QTL) and single nucleotide polymorphism (SNP) loci associated with soybean oil content were mapped. The results from both methods were comprehensively analyzed to mine oil content-related genes. Furthermore, haplotype analysis of new soybean varieties from Heilongjiang Province was conducted using marker information.

The main findings are as follows:

(1) Linkage analysis was performed using oil content data from 329 recombinant inbred lines (RILs) derived from the cross between the high-oil varieties Qihuang 34 and Dongsheng 16, evaluated over two years and two locations, combined with a genetic map constructed using 6,297 SLAF markers developed via simplified resequencing. Three major-effect QTL for oil content (qOC24-3-1, qOC23-20-1, and qOC23-20-2) were mapped. Notably, no previously reported soybean oil content-related gene loci exist near qOC23-20-2. By analyzing the average oil content of the RIL population across both years and locations combined with genotyping data, the genetic interaction effects among these three QTL were examined. The results indicated that qOC23-20-2 is the major-effect locus among them. Within the qOC23-20-2 region, three candidate genes (Glyma.20G237600, Glyma.20G238500, and Glyma.20G238800) were identified. Among these, Glyma.20G238500 is predicted to regulate lipid metabolism and is a candidate gene associated with oil synthesis.

(2) Genome-wide association analysis was conducted using oil content phenotypic data from the natural population of 341 accessions over two years, combined with 1,048,576 genome-wide SNP markers obtained via resequencing. A total of 119 significant SNP loci were detected. Multiple significant SNP loci associated with soybean seed oil content in both years were identified on chromosome 8. Candidate gene screening on chromosome 8 yielded four candidate genes (Glyma.08G110000, Glyma.08G117400, Glyma.08G117600, and Glyma.08G123500).

(3) Haplotype analysis using candidate gene information was performed on new soybean varieties from Heilongjiang Province, and relevant KASP markers were developed. The results showed that Glyma.08G117600 and Glyma.08G123500 significantly enhance oil content. Varieties harboring the oil-content-increasing genotypes at these loci are less prevalent among varieties adapted to the first accumulated temperature zone of Heilongjiang. Therefore, Glyma.08G117600 and Glyma.08G123500 are crucial for future breeding efforts to develop new high-oil soybean varieties in Heilongjiang Province.

Based on the above findings, this study identified 3 high-efficiency QTLs and 119 significant SNP sites, uncovering 7 candidate genes. Among these, Glyma.20G238500, Glyma.08G117600, and Glyma.08G123500 serve as crucial candidate genes for breeding high-oil soybean varieties. This research provides a theoretical basis and technical support for molecular marker-assisted selection of soybean oil-related traits.

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