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锦葵科作物SSR标记在玫瑰茄中的通用性分析

李云清 谢瑞真 李静 林荔辉 陈冀涵 方平平 陶爱芬

李云清,谢瑞真,李静,等. 锦葵科作物SSR标记在玫瑰茄中的通用性分析 [J]. 福建农业学报,2023,38(3):302−311 doi: 10.19303/j.issn.1008-0384.2023.03.006
引用本文: 李云清,谢瑞真,李静,等. 锦葵科作物SSR标记在玫瑰茄中的通用性分析 [J]. 福建农业学报,2023,38(3):302−311 doi: 10.19303/j.issn.1008-0384.2023.03.006
LI Y Q, XIE R Z, LI J, et al. Universality of Simple Sequence Repeat (SSR) Markers from Malvaceae plants to Roselle (Hibiscus sabdariffa L.) [J]. Fujian Journal of Agricultural Sciences,2023,38(3):302−311 doi: 10.19303/j.issn.1008-0384.2023.03.006
Citation: LI Y Q, XIE R Z, LI J, et al. Universality of Simple Sequence Repeat (SSR) Markers from Malvaceae plants to Roselle (Hibiscus sabdariffa L.) [J]. Fujian Journal of Agricultural Sciences,2023,38(3):302−311 doi: 10.19303/j.issn.1008-0384.2023.03.006

锦葵科作物SSR标记在玫瑰茄中的通用性分析

doi: 10.19303/j.issn.1008-0384.2023.03.006
基金项目: 国家现代农业产业技术体系建设专项(CARS-16);福建省南方麻类种质资源共享平台项目(PTJH20006);福建农林大学校内创新基金项目(KFA17164A、KFA19028A)
详细信息
    作者简介:

    李云清(1996−),女,研究方向:特色经济作物遗传育种(E-mail:lyq17853514030@163.com

    通讯作者:

    方平平(1968−),男,副教授,研究方向:麻类作物遗传育种研究(E-mail:fangpp1990@qq.com

    陶爱芬(1976−),女,副研究员,研究方向:麻类作物遗传育种研究(E-mail:281770126@ qq.com

  • 中图分类号: S563.9

Universality of Simple Sequence Repeat (SSR) Markers from Malvaceae plants to Roselle (Hibiscus sabdariffa L.)

  • 摘要:   目的  丰富玫瑰茄(Hibiscus sabdariffa L.)SSR分子标记的种类和数量,验证锦葵科作物SSR标记在玫瑰茄中的通用性,进而分析玫瑰茄的遗传多样性。  方法  选取锦葵科红麻、棉花、黄秋葵和木槿等4种作物共62对SSR引物(木槿17对,其他各15对),对8份不同来源的玫瑰茄种质进行多态性分析。  结果  62对SSR引物中有58对具有通用性,占比为93.5%;通用性引物中有48对具有多态性,占比为82.8%;这些引物共扩增出246条条带,其中多态性条带239条,占比97.2%。同时,棉花有14对SSR引物具有多态性,多态性引物比率最高,达86.7%;其次是木槿,有82.4%的引物具有多态性,且其引物多态信息含量值(PIC)最高,为0.685;而黄秋葵和红麻多态性引物比率较低,分别为73.3%和66.7%。另外,聚类分析结果表明,在相似系数 0.49处可将 8 份玫瑰茄种质分为3个各具特点的亚类,其中19FZ-76被单独聚为一类,表明这一种质与其他7份种质亲缘关系最远,遗传差异最大,聚类结果与其表型性状表现一致;而来自缅甸的19FZ-74和漳州本地种遗传相似系数最高,说明这两份种质的亲缘关系较近,遗传差异较小,表明我国福建种植的玫瑰茄品种与东南亚的品种有较近的血缘关系,有可能从东南亚引种而来。  结论  锦葵科作物的SSR引物在玫瑰茄中具有较好的通用性,筛选出的SSR标记可为玫瑰茄品种鉴定、亲缘关系分析、遗传图谱构建等遗传学研究提供可靠方法。
  • 图  1  3对不同引物在8份玫瑰茄种质中的扩增结果

    M:100 bp ladder marker;1:FAFU1;2:H150;3:19FZ-74;4:白桃K;5:漳州本地种;6:MG5;7:MG6;8:19FZ-76。引物1:HA-1;引物2:HA-2;引物3:HA-3。

    Figure  1.  Amplified 3 SSR primers for 8 roselle accessions

    M: 100 bp ladder marker; 1: FAFU1; 2: H150; 3: 19FZ-74; 4: Baitao K; 5: Zhangzhou native cultivar; 6: MG5; 7: MG6; 8:19FZ-76; Primer 1: HA-1; Primer 2: HA-2; Primer 3: HA-3.

    图  2  8份玫瑰茄种质基于SSR标记的聚类分析

    Figure  2.  SSR marker-based clustering on 8 roselle accessions

    表  1  供试玫瑰茄的名称、来源地及主要性状

    Table  1.   Names, origins and main phenotypes of roselle germplasms

    编号 No.名称
    Name
    来源地
    Origin
    主要性状
    Main phenotypes
    1 FAFU1 中国福建
    紫色肉质花萼,掌状浅裂叶,茎紫红色
    2 H150 中国湖南
    无肉质花萼,掌状全裂叶,茎紫红色
    3 19FZ-74 缅甸仰光
    无肉质花萼,掌状深裂叶,茎紫红色
    4 白桃K Baitao K
    中国广东
    绿色肉质花萼,掌状浅裂叶,茎浅绿色
    5 漳州本地种
    Zhangzhou native cultivar
    中国福建
    紫色肉质花萼,掌状深裂叶,茎紫红色
    6 MG5 中国福建
    红色肉质花萼,掌状深裂叶,茎紫红色
    7 MG6 中国福建
    紫色肉质花萼,掌状浅裂叶,茎紫红色
    8 19FZ-76 中国福建
    紫色肉质花萼,掌状全裂叶,茎紫红色
    下载: 导出CSV

    表  2  供试引物名称和序列信息

    Table  2.   Names and sequence information of SSR primers

    来源
    Origin
    名称
    Name
    引物上游序列(5′-3′)
    Foward primer sequence (5′-3′)
    引物下游序列(5′-3′)
    Reverse primer sequence(5′-3′)
    木槿 Rosemallow HA-1 TTGAACATAAACAAGCGG AAAACAAGTTGGGGAGG
    HA-2 CTGAATGCCAGAATGACT CAGGCGAAAGAGGAAGAT
    HA-3 ATCATTATCATCTTCGTTTC AAGGGACCAAAGTCTCAA
    HA-4 CACCAAATCCTGGAGAAG GCAAACGAGAATAATCAAAA
    HA-5 GCGTGGATGTTCTTTCTT TCGAACCCTATGGATGTA
    HA-6 GAACAAGCCTGTCACTAA CACAAACCGATTTACGAT
    HA-7 CAGCAGTTAGAGCAGGAGGT TTCGGACATGAGTATGGGAT
    HA-8 CACTTCCACGAAGCTCTTAC GGAGATAAACAGAAAAGGGTA
    HA-9 TATGGGTTTAGTGCCTGTAT TAGGTTGCTTGAATCTTTTC
    HA-10 CCCAAACCTCTATCATCT ATATCCCTTAGTTCTGCT
    HA-11 CACCAAATCCTGGAGAAGTA GGCAAACGAGAATAATCAAA
    HA-12 AAGGAGAAGCCAAGGTGAAA GACAAACCCACATACAGGAA
    HA-13 ACTTTTATCGTATAGACCAG GAACACCTTTATTTCAGTGT
    HA-14 GAAATGGCAAGGTTTTAGAT CTCAACTTTTGTGATGTGGC
    HA-15 CAGCCACAATCCTCCTAACT GAAGGGTAACTTGTTTCACG
    HDAT-1 CCCTTCAAGTGCTCCTCT TCAATTCACCTTCCGTACCC
    HDAT-2 TGTCAAGCTGTCAAGGGTGA CCGATCCGTGTTTTTCAAGT
    红麻 Kenaf HcES0010 AGCACATTCCCGGTTGCTAA TGCAGCAAAGCAATGTAGGC
    HcES0011 GCTAAGCTGGGTTTCTGGGA CCCAACAGCTTCTATTCACTGC
    HcES0012 CCCCCTCCATTTGGGTTTCT AACCCAGCAATCCCTTGGTT
    红麻 Kenaf HcES0014 CGTGCATGTCGTGAAATCCG ATTACAAAGTGGGCAGGCGA
    HcES0015 TGGGAAAGGGATTGGAACCA ACAATTCCGTCACCACCTCC
    HcES0020 CACGTGTGACCACAGAGCTA AACAAAACATGCAGCTCCCG
    HcES0041 ATCCAATCCAATCCAAACTCCT TCAGGTATAAGTCGGGGCGT
    HcES0042 CCCCATTGACTCGAGGCTTT AGCCTTTACAATACTTAGTCCGGT
    HcES0060 AGGTGTCTGGTGACTGGTCA GTGCAAGAACTGGAGTTGCG
    HcES0063 AGTTGGGATCTGCTGACACG CTGAACCGAAATCAAGGCACA
    HcES0064 CCACTAGAAGTTTATCGTTGGCC AGAGTATGTGCACGGGAAGC
    HcES0070 AGGCTGTTGCTGAGCTGAAT CGCCGTTAGCAATAGCAACC
    HcES0075 CAGGAAGGAAATGGGCAGGA ATCGACCGAAACTCGAGAGC
    HcES0085 AGCCATCCCATTGCAAGACA TCACTGCATCACCCTTCCAA
    HcES0087 CCTCTGATGCATCCGCTTCT ATCTCCAGGGTGTTGCGTTT
    C2-0004C TTTGTTTTGCGTTCCTTTA TCCGACAATGCCTTACAAG
    C2-0005B CCCCATTCCTACTCATCC CACAGAAAGGTGCTCATGC
    CGR5501 TCTCTCTTGCTGGTCACGAA TGCCAAATACCCAAATCCAT
    CGR5503 GCTGCTTCCATGCCATTATT GGGTCGCTTTGTAAGTGAATG
    CGR5506 CAGCAACCACAATTCGATCA GAAGTTGCTGTTGGGAAGGA
    棉花 Cotton CGR5508 CAACTTTCCGAGCTGGATTC TGATCGAGGAATGAAAGCAA
    CGR5645 GAGCGGAGAGTCCGGTTT CCCAAACGAATCAAAGATGG
    DC30003 AGGAGGGAAAGAGTGGTG CCTCCTCACATCCAATCA
    DC30005 ATGAGAAACGGTGTCGAA TTGACCGAATACTCCCCT
    DPL0840 GAGTCGTTGCCGCTGTTTA GCTACGACTCGATGTTACGG
    SHIN0733 GCTTTGCCTTCGGTTCATT GGACTTCGCTTTATGAATGCTT
    SHIN0745 GCACCGAGTCTCCTATGCTC GGACCCTCAAACTTGTATTACACT
    CIR102 TAACAACTGGATGAGATGA CATAACTGCAAAGGAGAA
    DC20067 ATGCAAACCATAAACATCT TGGGTTTGTGTGCTATCT
    DPL0063 GTTCATACATATGGAGAGGCAGC ATGCTTCTCACATGGCTACTCTTT
    黄秋葵 Okra HeSSR002 CGCTGCTCCAGACATAACAA TCTTTCTCTCCGAACCTCCA
    HeSSR003 GGCGATCAAATTAGAAACCAG GTCACGCTGTAGCTCTCGAA
    HeSSR009 GAAGAACAAGGCAAGGCAAG AACATCTCCAAGCTCCCTCA
    HeSSR011 TGCAGGCAAATCAACTGAAAG GTTTCCATTCGACTCGGGTA
    HeSSR017 ACTGCCACTCAACAGCAATG TGCGACAATATTTCCAGCAG
    HeSSR020 TTCTCACTGCAGCAACAAGG TGCCACTTCTCTGCCTCTTT
    HeSSR023 CAAGGAGACAATGCAGGGTT ATTCGATACCCAAAGCTCCC
    HeSSR030 GAGATCAAGAACGTGTGGCA AATCTTCGTTGGTTTCGGTG
    HeSSR033 TAAGAGGCGTGAAAGGAAGC AAGCTCAACCTTTACGGCAA
    HeSSR036 GTGGGTTCAAACTTTGGGTGAAG CAGCAAGGTTATTACACTTTACA
    HeSSR037 ACGTCATCCTCATTAACGGC TTCCTTTCCGACAATTCGAC
    HeSSR040 CGTTTGAACGCTACCCCATA CCCGAAACCACCTAACTCAA
    HeSSR041 GCGAATCTTTTGGTATCGGA ATGGATGATTTGGTGGGATG
    HeSSR043 GCTGCAGCTAAAACGCTTCT CGTAGTCCATGAGGGCAAAT
    HeSSR046 GAATGGAAGCCAAAGGATCAT GGAATGAATTTTATGATGATGG
    下载: 导出CSV

    表  3  不同来源的SSR引物在玫瑰茄中的扩增情况

    Table  3.   Amplified SSR primers in roselle

    标记来源
    Origins
    标记总数
    Number of
    primers
    引物通用率
    Transferability/
    %
    多态性引物比率
    Ratio of polymorphic
    primers/%
    多态性条带比率
    Ratio of polymorphic
    bands/%
    有效条带数量范围
    Number range of
    effective bands
    平均扩增条带
    Average amplified
    bands
    木槿 Rosemallow 17 94.1 82.4 100 3~8 5.5
    红麻 Kenaf 15 86.7 66.7 98.0 1~8 4.5
    黄秋葵 kra 15 100 73.3 96.0 2~8 5
    棉花 Cotton 15 93.3 86.7 93.8 1~9 5
    总数 Total 62 93.5 82.8 97.2
    下载: 导出CSV

    表  4  8份玫瑰茄种质的遗传多样性信息

    Table  4.   Genetic diversity of 8 roselle accessions

    来源
    Origin
    引物编号
    Primer code
    等位基因数
    Na
    有效等位基因数
    Ne
    Shannon多样性指数
    I
    观测杂合度
    Ho
    期望杂合度
    He
    平均Nei's多样性指数
    H
    多态信息含量
    PIC
    木槿 Rosemallow HA-1 6 4.923 1.684 0.875 0.800 0.797 0.768
    HA-2 8 6.737 1.981 0.750 0.850 0.852 0.834
    HA-3 3 2.085 0.892 0.750 0.908 0.520 0.464
    HA-4 6 4.414 1.635 0.429 0.560 0.773 0.744
    HA-5 6 3.657 1.509 0.625 0.825 0.727 0.691
    HA-7 6 4.167 1.609 0.571 0.659 0.760 0.730
    HA-8 4 1.969 0.951 0.625 0.725 0.492 0.458
    HA-9 7 5.333 1.787 0.625 0.775 0.813 0.787
    HA-10 6 4.741 1.646 0.625 0.867 0.789 0.757
    HA-11 7 4.571 1.721 0.625 0.842 0.781 0.755
    HA-12 8 5.818 1.923 0.125 0.525 0.828 0.809
    HA-13 3 2.133 0.900 0.500 0.833 0.531 0.468
    HA-14 5 3.200 1.332 0.500 0.567 0.688 0.636
    HA-15 5 3.657 1.455 0.625 0.883 0.727 0.690
    红麻 Kenaf HcES011 4 2.612 1.157 0.375 0.675 0.617 0.572
    HcES012 2 1.133 0.234 0.400 0.844 0.117 0.110
    HcES014 8 5.565 1.890 0.125 0.125 0.820 0.799
    HcES015 6 4.741 1.663 0.625 0.875 0.789 0.759
    HcES020 5 2.000 1.037 0.875 0.842 0.500 0.474
    HcES041 6 2.370 1.247 0.375 0.533 0.578 0.553
    HcES060 7 6.533 1.909 0.375 0.658 0.847 0.828
    HcES070 5 2.000 1.037 0.625 0.617 0.500 0.474
    HcES075 5 4.000 1.494 0.286 0.912 0.750 0.712
    HcES087 3 2.032 0.831 0.500 0.533 0.508 0.428
    黄秋葵 Okra HeSSR002 3 2.246 0.882 1.000 0.592 0.555 0.456
    HeSSR003 5 3.122 1.354 0.000 0.800 0.680 0.641
    HeSSR009 6 2.032 1.124 0.625 0.542 0.508 0.489
    HeSSR017 3 1.684 0.736 0.250 0.233 0.406 0.371
    HeSSR020 8 5.765 1.909 0.000 0.000 0.827 0.806
    HeSSR023 3 1.684 0.736 0.714 0.890 0.406 0.371
    HeSSR033 4 2.941 1.221 0.000 0.433 0.660 0.610
    HeSSR040 4 3.600 1.330 0.500 0.433 0.722 0.671
    HeSSR041 8 7.111 2.014 0.500 0.450 0.859 0.843
    HeSSR043 3 1.343 0.509 0.000 0.788 0.255 0.240
    HeSSR046 3 2.723 1.043 0.375 0.917 0.633 0.556
    棉花 Cotton C20005B 5 4.000 1.494 0.286 0.275 0.750 0.712
    CGR5501 6 2.370 1.247 0.375 0.775 0.578 0.553
    CGR5503 9 7.000 2.069 0.625 0.617 0.857 0.842
    CGR5506 3 1.471 0.602 0.571 0.923 0.320 0.294
    CGR5508 2 1.882 0.662 0.125 0.342 0.469 0.359
    CGR564 4 2.579 1.091 0.250 0.800 0.612 0.541
    SHIN734 5 1.730 0.909 0.750 0.900 0.422 0.404
    SHIN745 2 1.280 0.377 0.750 0.800 0.219 0.195
    DC20067 7 3.368 1.560 0.200 0.733 0.703 0.678
    DC30005 7 6.400 1.895 0.750 0.500 0.844 0.824
    DPL0840 3 3.000 1.099 0.625 0.542 0.667 0.593
    CIR102 7 4.000 1.661 0.625 0.750 0.750 0.725
    C2-0004C 4 2.246 1.041 1.000 0.733 0.555 0.511
    总计 Total 245 165.969 62.088 23.707 32.002 30.360 28.583
    平均值 Average value 5 3.458 1.293 0.494 0.667 0.633 0.595
    下载: 导出CSV
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出版历程
  • 收稿日期:  2022-11-11
  • 录用日期:  2022-11-11
  • 修回日期:  2023-01-17
  • 网络出版日期:  2023-03-28
  • 刊出日期:  2023-03-28

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