番茄品种KASP标记鉴定技术体系的构建与应用

doi:10.3864/j.issn.0578-1752.2025.22.014

摘要/Abstract

摘要：

【目的】筛选KASP核心标记，建立番茄高通量分子鉴定体系，为番茄品种鉴定、纯度鉴定和新品种权保护等提供技术支撑。【方法】基于番茄高代自交系重测序数据，筛选覆盖全基因组的高质量SNP位点，并将其转化为KASP标记。利用从不同育种单位和公司收集的360个番茄品种验证开发的KASP标记，根据基因分型数据结果，筛选出一套多态性信息含量（PIC）高、稳定性好且在染色体上相对均匀分布的KASP核心标记组合，并对收集的番茄品种进行聚类分析和构建DNA指纹图谱，同时进行番茄品种真实性鉴定和种子纯度鉴定应用。【结果】基于300余份番茄材料重测序数据分析获得的SNP位点，从中筛选出600个用于标记开发，平均每条染色体50个。最终，517个成功转化为KASP标记，转化率为86.2%。根据染色体上的物理距离，每条优选10—12个，总计128个标记，使用不同类型的番茄品种对其进行验证和筛选。最终筛选出一套含有68组引物的核心标记，平均每条染色体4—8个。利用这68个核心标记对360个番茄品种进行基因型分析，并构建DNA指纹图谱，聚类分析结果显示，品种间相似系数为0.32—1.00。依据普通鲜食番茄、口感番茄、樱桃番茄和加工番茄4种类型，分别精简为含有48、32、36和36个核心标记的组合，其PIC值均大于0.35，这些核心标记可将不同品种的番茄进行有效区分。为了验证标记的适用性，利用核心标记组合对2个口感番茄品种进行真实性鉴定，发现二者有2个标记存在差异，表明这两个品种并不相同。另外，选择在亲本间具有多态性的2个标记对番茄品种‘品番4031’的F₁杂交种子纯度进行鉴定，结果显示，种子纯度均为100%，与田间鉴定结果吻合。【结论】获得的KASP标记可用于番茄的遗传多样性分析、指纹图谱构建、品种真实性鉴定和杂交种纯度鉴定。

关键词: 番茄, KASP, 核心标记, 指纹图谱, 品种鉴定

Abstract:

【Objective】To screen KASP core markers and establish a high-throughput molecular identification system for tomato, providing technical support for tomato variety authentication, purity testing, and plant variety protection. 【Method】Based on re-sequencing data of tomato inbred lines, high-quality SNP loci covering the whole genome were selected for KASP markers design. A total of 360 tomato varieties, collected from different breeding research institutions and private companies, were used to validate the designed KASP markers. According to genotyping data, a set of KASP core markers with high polymorphism information content (PIC), excellent stability, and relatively uniform distribution across chromosomes was screened. Cluster analysis and DNA fingerprinting of the collected tomato varieties were performed, and the core markers were applied to authenticate variety authenticity and assess seed purity. 【Result】Utilizing the SNPs obtained from re-sequencing data analysis of over 300 tomato materials, 600 SNP loci were selected for marker development, averaging 50 per chromosome. Among these, 517 were successfully converted into KASP markers, with a conversion success rate of 86.2%. Based on physical distances on chromosomes, 10-12 markers per chromosome were prioritized, totaling 128 markers, which were validated and refined using diverse tomato varieties. Ultimately, 68 core primer pairs were selected, averaging 4-8 per chromosome. Genotyping analysis of 360 tomato varieties using these 68 core markers revealed similarity coefficients ranging from 0.32 to 1.00 in cluster analysis. For four tomato types (fresh tomatoes, flavor tomatoes, cherry tomatoes and processing tomatoes), the core marker sets were streamlined to 48, 32, 36, and 36 markers, respectively, with PIC values all exceeding 0.35. These markers effectively distinguished different varieties. To validate marker applicability, two flavor tomato varieties were authenticated using core markers, revealing two differing loci, confirming their distinctness. Additionally, two polymorphic markers between parental lines were used to assess the purity of hybrid seeds of Pinfan 4031, yielding 100% purity, consistent with field evaluation results. 【Conclusion】The developed KASP markers are applicable for tomato genetic diversity analysis, DNA fingerprinting, variety authenticity testing, and hybrid seed purity evaluation.

Key words: tomato, KASP, core markers, fingerprint, variety identification

苏晓梅, 杨宗辉, 刘淑梅, 张宗杰, 吕宏君, 侯丽霞. 番茄品种KASP标记鉴定技术体系的构建与应用[J]. 中国农业科学, 2025, 58(22): 4746-4756.

SU XiaoMei, YANG ZongHui, LIU ShuMei, ZHANG ZongJie, LÜ HongJun, HOU LiXia. Development and Application of A KASP Marker-Based Identification System for Tomato Varieties[J]. Scientia Agricultura Sinica, 2025, 58(22): 4746-4756.

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