| 刘小飞 1,2，于 波 1，任桂萍 3，余超然 2,4，刘 冲 2,5，孙映波 1，钟荣辉 1，冯恩友 2.基于 SSR 标记的 12 个非洲菊品种指纹图谱构建及杂交 F1 后代鉴定[J].广东农业科学,2023,50(9):16-24 |
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基于 SSR 标记的 12 个非洲菊品种指纹图谱构建及杂交 F1 后代鉴定 |
| Construction of Fingerprint Map of 12 Varieties Based on SSR Markers and Identification of Hybrid F1 Progenies in Gerbera hybrida |
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| DOI：10.16768/j.issn.1004-874X.2023.09.002 |
| 中文关键词: 非洲菊 SSR 毛细管电泳 亲缘关系分析 指纹图谱 杂种鉴定 |
| 英文关键词: Gerbera hybrida SSR capillary electrophoresis relationship analysis fingerprint map identification of hybrida |
| 基金项目:广东省科技专项资金（“大专项 + 任务清单”）项目（花卉优质高效繁育技术服务团队）；湛江市农业科学研究院博士工作站项目（非洲菊种质资源收集与创新利用）；广东省农业科学院协同创新中心项目（XT202212）；广东省农业科学院学科团队建设项目（202128TD） |
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| 中文摘要: |
| 【目的】基于 SSR 分子标记对 12 份非洲菊种质进行亲缘关系分析及部分种质杂交 F1 后代真假杂种鉴定，以期为非洲菊种质资源鉴评及创新利用提供技术支撑。【方法】收集 12 个非洲菊种质资源，提取其基因组 DNA 后，利用 50 对 SSR 引物进行 PCR 扩增，通过检测相关多态性结果进行聚类分析和指纹图谱构建，以及部分品种杂交 F1 后代鉴定。【结果】18 对引物在 12 份非洲菊材料中共扩增出 74 个多态性等位变异片段，平均每对引物扩增出多态性等位变异片段 4.1 个。利用 74 个等位位点，构建了 12 份非洲菊种质资源的系统进化树，UPGMA 聚类分析结果显示，当遗传系数为 0.75 时，可将受测的 12 份种质分为 5 组，Ⅰ组为‘云南红’‘大088’‘情人’，Ⅱ组为‘粉佳人’‘粉球’‘真爱’和‘福娃’，Ⅲ组为‘绣色’‘淑女’和‘黄球’， Ⅳ组和Ⅴ组均只包含 1 个品种，分别是‘紫水晶’和‘深圳 5 号’。筛选出 GHSSR-1、GHSSR-18、GHSSR-20和 GHSSR-21 等 4 对引物，可完全区分 12 份种质；其中，后 3 对引物鉴定出兼具双亲特异位点的单株分别是Gh-5、Gh-4 和 Gh-1，单独 1 对引物的鉴定率为 33%。且受检的 3 个单株均为真杂种，真杂种率为 100%。【结论】成功构建 12 份非洲菊品种的指纹图谱，综合 3 对多态性较好的引物鉴定出 3 株非洲菊杂交 F1 后代实生苗为真杂种，为非洲菊资源鉴评和创新利用提供可靠的 SSR 标记引物。 |
| 英文摘要: |
| 【Objective】Based on the method of SSR molecular markers, this study conducted phylogenetic analysis on 12 germplasms and identified true and false hybrids from F1 hybrid progenies of some germplasms in Gerbera hybrida, in order to offer a technological support for the evaluation and innovative utilization of germplasm resources in G. hybrida. 【Method】 Twelve Gerbera hybrida germplasm resources were collected and their genomic DNA was extracted. PCR amplification was performed using 50 pairs of SSR primers. Cluster analysis and fingerprint construction were conducted by detecting the results of related polymorphisms, as well as identification of hybrid F1 offspring of some varieties.【Result】A total of 74 polymorphic alleles were amplified by 18 pairs of primers in 12 varieties of G. hybrida, with an average of 4.1 polymorphic alleles amplified by each pair of primers. A phylogenetic tree of 12 germplasm resources was constructed using 74 alleles in G. hybrida. UPGMA clustering analysis results showed that when the genetic coefficient was 0.75, the tested 12 germplasm resources could be divided into 5 groups, with Group I including ‘Yunnanhong’, ‘Da088’, and ‘Qingren’. Group II including ‘Fenjiaren’, ‘Fenqiu’, ‘Zhenai’, and ‘Fuwa’. Group III including ‘Xiuse’, ‘Shunv’, and ‘Huangqiu’, both Group IV and Group V containing only one variety, namely ‘Zishuijing’ and ‘Shenzhen 5’, respectively. Four pairs of primers were screened, including GHSSR-1, GHSSR-18, GHSSR-20, and GHSSR-21, which can completely distinguish the 12 germplasms with each other. Among them, the last three pairs of primers identified lines with parental specific heterotopies as Gh-5, Gh-4, and Gh-1, respectively. The identification rate of a single pair of primers was 33%. All three tested individual plants are true hybrids, with a true hybrid rate of 100%.【Conclusion】Twelve fingerprint maps of G.hybrida varieties were successfully constructed. At the same time, 3 hybrid F1 offspring seedlings were identified as true hybrids, providing dependable SSR marker primers for the evaluation and innovative utilization of resources in G. hybrida. |
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