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园艺学报 ›› 2023, Vol. 50 ›› Issue (2): 265-278.doi: 10.16420/j.issn.0513-353x.2021-1008

• 研究论文 • 上一篇    下一篇

石榴品种SSR指纹图谱构建及杂种父本鉴定

王瑞1,*, 洪文娟1,*, 罗华2, 赵丽娜2, 陈颖2, 王君1,**()   

  1. 1.北京林业大学生物科学与技术学院,林木育种与生态修复国家工程研究中心,林木花卉遗传育种教育部重点实验室,树木花卉育种生物工程国家林业和草原局重点实验室,北京 100083
    2.枣庄市石榴研究中心,山东枣庄 277300
  • 收稿日期:2022-08-11 修回日期:2022-12-02 出版日期:2023-02-25 发布日期:2023-03-06
  • 通讯作者: **(E-mail:wangjun@bjfu.edu.cn
  • 作者简介:第一联系人:

    *共同第一作者

  • 基金资助:
    林业行业标准制修订项目(2020-LY-061);枣庄市石榴国家林木种质资源库良种繁育补助项目;枣庄市自主创新及成果转化计划项目(枣科字〔2020〕75号)

Construction of SSR Fingerprints of Pomegranate Cultivars and Male Parent Identification of Hybrids

WANG Rui1,*, HONG Wenjuan1,*, LUO Hua2, ZHAO Lina2, CHEN Ying2, WANG Jun1,**()   

  1. 1. National Engineering Research Center of Tree Breeding and Ecological Restoration,Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants of Ministry of Education,The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of National Forestry and Grassland Administration,College of Biological Sciences and Biotechnology,Beijing Forestry University,Beijing 100083,China
    2. National Germplasm Bank of Pomegranate,Zaozhuang,Shandong 277300,China
  • Received:2022-08-11 Revised:2022-12-02 Online:2023-02-25 Published:2023-03-06
  • Contact: **(E-mail:wangjun@bjfu.edu.cn

摘要:

以枣庄市石榴国家林木种质资源库中收集的168个石榴品种为材料,利用SSR分子标记技术构建了石榴品种资源指纹图谱,并基于指纹图谱信息,应用最大似然值法(LOD)对品种‘秋艳’和‘会理青皮软籽’的自然授粉实生苗进行了杂种筛选及父本初步鉴定。22对SSR引物从168个石榴品种样品中共扩增到76个等位标记位点,其中多态性位点58个,多态性为76.3%。平均等位位点数(Na)、有效等位位点数(Ne)、观测杂合度(Ho)、期望杂合度(He)分别为3.455、2.269、0.362、0.538,Shannon’s信息指数(I)平均值为0.897,多态性信息含量(PIC)介于0.330 ~ 0.692之间,平均PIC值为0.462。利用22对SSR引物可将166个石榴品种区分开,另有‘大满天红甜’和‘小满天红甜’2个品种未有效区分,有同物异名的可能;在此基础上构建了166个品种的特异指纹代码及指纹图谱二维码。基于指纹图谱信息,利用SSR标记从‘秋艳’的30个自然授粉子代实生苗中筛选到1个真杂种Q-1,疑似父本为‘峄城丰产马牙’,LOD值为5.96;从‘会理青皮软籽’的118个自然授粉子代实生苗中筛选到6个真杂种H-4、H-49、H-51、H-87、H-94和H-103,疑似父本分别为‘四川黄皮酸’‘突尼斯软籽’‘蒙自红花白皮’‘四川黄皮酸’‘突尼斯软籽’及‘蒙自红花白皮’。

关键词: 石榴, 品种资源, SSR指纹图谱, 遗传多样性, 父本鉴定

Abstract:

Young leaves of 168 pomegranate cultivars collected from the National Forest Germplasm Resource Repository for Pomegranate,Zaozhuang City,Shandong,China were used to construct their fingerprints using SSR molecular marker technique. Based on the fingerprint information,the maximum likelihood method(LOD)was applied to hybrid screening in open-pollinated progenies of the cultivars ‘Qiuyan’and‘Huili Qingpi Ruanzi’and identification of suspected male parent. The results showed that a total of 76 alleles were amplified from 22 pairs of SSR primers in the 168 cultivar samples,including 58 polymorphic loci(accounting for 76.3%). The average number of alleles per locus(Na),effective number of alleles(Ne),observed heterozygosity(Ho)and expected heterozygosity(He)was 3.455,2.269,0.362 and 0.538,respectively. The average Shannon’s information index(I)was 0.897,the polymorphism information content(PIC)of each marker ranged from 0.330 to 0.692,with an average of 0.462. 166 pomegranate cultivar samples were distinguished by the 22 pairs of SSR primers. However,‘Damantianhongtian’and‘Xiaomantianhongtian’were not effectively distinguished,which might be synonymous. Using the allelic information,a database containing the specific fingerprint code and two-dimensional fingerprint code for each cultivar was constructed. One true hybrid Q-1 was screened from 30 offspring of open-pollinated‘Qiuyan’by using the fingerprint information. Its suspected male parent is‘Yicheng Fengchan Maya’,with the LOD value of 5.96. Six true hybrids,H-4,H-49,H-51,H-87,H-94 and H-103,were screened from 118 offspring of open-pollinated‘Huili Qingpi Ruanzi’. Their suspected male parents were‘Sichuan Huangpisuan’‘Tunisian Ruanzi’‘Mengzi Honghua Baipi’‘Sichuan Huangpisuan’‘Tunisian Ruanzi’and‘Mengzi Honghua Baipi’,respectively.

Key words: Punica granatum, cultivar resource, SSR fingerprint, genetic diversity, male parent identification

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