基于兰州百合基因组对龙牙与兰州百合杂交后代的SSR分子标记开发及真实性鉴定OA
Development and Authenticity Identification of SSR Molecular Markers from Hybrid Progenies of Lilium brownii var.viridulum and Lilium davidii var.unicolor Based on the Genome of Lilium davidii var.unicolor
[目的]利用兰州百合全基因组开发食用百合 SSR标记,结合切柱头、分子鉴定等相关技术建立从远缘杂交到子代鉴定的完整技术体系,为食用百合杂交育种提供快速准确的鉴定方法.[方法]整合正反交授粉、切柱头授粉、胚挽救、生物信息学、SSR分子标记及细胞学鉴定等技术,通过比对兰州百合基因组和龙牙百合转录组数据筛选 SSR位点,从 96对 SSR引物中筛选重复性好并具有亲本特异性条带的引物,并对34株 F1 代杂交苗进行鉴定.[结果]共筛选出 5 244个 SSR位点,重复类型涵盖单核苷酸至六核苷酸,以单核苷酸(31.10%)和二核苷酸重复(42.32%)为主,数量较多的基元分别为 A/T和 TA/AT、AT/AT,且 10次重复频次最高(共计 817个,占 SSR位点总数的 15.58%).从 96对 SSR引物中筛选出 5对(BHSSR002、BHSSR005、BHSSR016、BHSSR074、BHSSR096)能产生互补特异性条带的引物,其中,4对引物(BHSSR002、BHSSR005、BHSSR016、BHSSR096)对 34株 F1 代杂交苗的综合鉴定效率达 94.12%.核型分析显示,父本特异性"V"型染色体在杂交子代中稳定遗传.[结论]兰州百合与龙牙百合共有的 SSR位点以单核苷酸和二核苷酸重复为主,A/T、TA/AT为优势基元,10次重复单元出现频率最高;筛选获得的 4对 SSR引物可高效鉴定杂交后代真实性;父本特征性"V"型染色体可作为细胞学鉴定标记.
[Objective]The integrity technical system from distant hybridization to progeny identification for edible lily was established by the related techniques including SSR markers developed from the complete genome of Lilium davidii var.unicolor,cutting stigma and molecular identification to provide a fast and accurate identification method for cross breeding of edible lily.[Method]The primers with good repeatability and specific parental bands were selected from 96 pairs of SSR primers by comparing L.davidii var.unicolor genome and L.brownii var.viridulum transcriptome data,and 34 F1 hybrid seedlings were identified by the integrating techniques including reciprocal cross-pollination,pollination by cutting stigmas,embryo rescue,bioinformatics,SSR molecular marker and cytological identification.[Result]A total of 5 244 SSR loci were identified,and the repeat types ranged from mono-nucleotide to hexa-nucleotide.The proportion of mono-nucleotide and di-nucleotide was 31.10%and 42.32%,respectively.The most abundant motifs were A/T,TA/AT and AT/AT,and the proportion of 817 SSR loci with the highest frequency of 10 repeats accounted for 15.58%of total SSR loci.5 pairs of SSR primers(BHSSR002,BHSSR005,BHSSR016,BHSSR074 and BHSSR096)with complementary specific bands were selected from 96 pairs of SSR primers.4 pairs of primers(BHSSR002,BHSSR005,BHSSR016 and BHSSR096)had a comprehensive identification efficiency of 94.12%for 34 F1 hybrid plants.Karyotype analysis revealed that the paternal-specific V-shaped chromosome was stably inherited in the hybrid progeny.[Conclusion]The SSR loci shared by L.davidii var.unicolor genome and L.brownii var.viridulum are mainly composed of mono-nucleotide and di-nucleotide repeats.A/T and TA/AT are the dominant motifs,and the 10-repeated motifs have the highest occurrence frequency.The screened 4 pairs of SSR primers can efficiently identify the authenticity of hybrid progenies.The characteristic V-shaped chromosome of the male parent can be used as a cytological identification marker.
王维泽;周舒婷;石乐娟;唐玲涵;晋宇轩;杜致辉;张朝君;陈之林
贵州省园艺研究所/贵州省园艺工程技术研究中心,贵州 贵阳 550006贵州省天然产物研究中心,贵州 贵阳 550014贵州省园艺研究所/贵州省园艺工程技术研究中心,贵州 贵阳 550006贵州省园艺研究所/贵州省园艺工程技术研究中心,贵州 贵阳 550006贵州省园艺研究所/贵州省园艺工程技术研究中心,贵州 贵阳 550006贵州省园艺研究所/贵州省园艺工程技术研究中心,贵州 贵阳 550006贵州省园艺研究所/贵州省园艺工程技术研究中心,贵州 贵阳 550006贵州省园艺研究所/贵州省园艺工程技术研究中心,贵州 贵阳 550006
农业科技
百合兰州百合龙牙百合杂种群体鉴定SSR分子标记基因组细胞学鉴定
Lilium sppLilium davidii var.unicolorLilium brownii var.viridulumhybrid identificationSSR molecular markergenomecytological identification
《贵州农业科学》 2026 (10)
119-132,14
贵州省农业科学院青年基金(黔农科院青年基金[2022]11号)贵州省科技计划项目(黔科合支撑[2023]一般460)(黔科合服企[2022]005)
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