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芍药属组内组间杂交及部分后代核型分析与SSR鉴定

刘建鑫 杨柳慧 魏冬霞 于晓南

刘建鑫, 杨柳慧, 魏冬霞, 于晓南. 芍药属组内组间杂交及部分后代核型分析与SSR鉴定[J]. 北京林业大学学报, 2017, 39(4): 72-78. doi: 10.13332/j.1000-1522.20160181
引用本文: 刘建鑫, 杨柳慧, 魏冬霞, 于晓南. 芍药属组内组间杂交及部分后代核型分析与SSR鉴定[J]. 北京林业大学学报, 2017, 39(4): 72-78. doi: 10.13332/j.1000-1522.20160181
LIU Jian-xin, YANG Liu-hui, WEI Dong-xia, YU Xiao-nan. Intrasectional and intersectional cross breeding of Paeonia and karyotype analysis and SSR identification of some hybrids[J]. Journal of Beijing Forestry University, 2017, 39(4): 72-78. doi: 10.13332/j.1000-1522.20160181
Citation: LIU Jian-xin, YANG Liu-hui, WEI Dong-xia, YU Xiao-nan. Intrasectional and intersectional cross breeding of Paeonia and karyotype analysis and SSR identification of some hybrids[J]. Journal of Beijing Forestry University, 2017, 39(4): 72-78. doi: 10.13332/j.1000-1522.20160181

芍药属组内组间杂交及部分后代核型分析与SSR鉴定

doi: 10.13332/j.1000-1522.20160181
基金项目: 

国家自然科学基金项目 31400591

北京市教育委员会科学研究与研究生培养共建项目 BLCXY201629

详细信息
    作者简介:

    刘建鑫。主要研究方向:芍药资源与育种。Email:jianxinliu@hotmail.com  地址:100083  北京市海淀区清华东路35号北京林业大学园林学院

    责任作者:

    于晓南,博士,教授。主要研究方向:园林植物资源与育种。Email:yuxiaonan626@126.com  地址:同上

  • 中图分类号: S682.1+2

Intrasectional and intersectional cross breeding of Paeonia and karyotype analysis and SSR identification of some hybrids

  • 摘要: 以芍药品种‘朱砂判’为母本,分别与6个芍药组品种和4个牡丹组品种进行杂交,并综合利用核型分析和SSR分子标记法对部分杂交后代进行杂种鉴定,筛选出亲和性较好的远缘杂交组合,并确定杂交后代早期鉴定方法,为芍药属新品种培育提供参考。结果表明:1)芍药组内近缘杂交亲和性好,平均结实率均在5粒/朵以上;芍药组内远缘杂交亲和性较差,但仍有一定结实;芍药组间远缘杂交表现出不亲和性,仅杂交组合‘朱砂判’בDao Da Chen’有较少结实。2)核型分析表明:组内远缘杂交后代ZC4、ZC5、ZC6、ZC7为三倍体(2n=3x=15);从50对芍药SSR引物中筛选出7对多态性引物进行扩增,结果显示杂交后代ZC4、ZC5、ZC6、ZC7含有父母本特异性条带,结合核型分析证明ZC4、ZC5、ZC6、ZC7为真杂种。以上结果表明:组内远缘杂交中‘朱砂判’与‘Garden Peace’、‘Pink Teacup’、‘Cream Delight’杂交均有一定结实,组间远缘杂交中‘朱砂判’בDao Da Chen’可打破杂交不亲和性;SSR分子标记技术可用于芍药属植物杂交后代的早期鉴定。

     

  • 图  1  ‘朱砂判’בCream Delight’杂种后代早期形态图

    Figure  1.  Early morphology of 'Zhu Sha Pan'× 'Cream Delight' hybrids

    图  2  亲本及杂交后代的染色体核型和模式图(‘朱砂判’בCream Delight’)

    Figure  2.  Chromosome karyotypes and idiograms of hybrids and their parents ('Zhu Sha Pan'× 'Cream Delight')

    图  3  引物S3、55、S16荧光检测图

    荧光检测图顺序对应表 5中的编号

    Figure  3.  Fluorescence detection figures by primer S3, 55, S16

    No. corresponding to the No. in Tab. 5

    表  1  父本基本信息

    Table  1.   Basic information of male parent

    杂交类型
    Hybrid type
    父本
    Male parent
    品种群
    Cultivar group
    花型
    Flower type
    花色
    Flower color
    组内近缘杂交
    Hybridization in Lactiflora group
    ‘Kansas’中国芍药品种群
    Lactiflora group
    皇冠型
    Crown type
    红色
    Red
    ‘Taff’中国芍药品种群Lactiflora group皇冠型
    Crown type
    粉色
    Pink
    ‘Karl Rosenfield’中国芍药品种群
    Lactiflora group
    菊花型
    Chrysanthemum type
    紫红色
    Purple
    组内远缘杂交Interspecific hybridization in sect. Paeonia‘Garden Peace’杂种芍药品种群Hybrid group单瓣型
    Single type
    白色
    White
    ‘Pink Teacup’杂种芍药品种群Hybrid group单瓣型
    Single type
    奶油粉
    Cream pink
    ‘Cream Delight’杂种芍药品种群Hybrid group单瓣型
    Single type
    奶油色
    Cream
    组间远缘杂交
    Intersectional hybridization
    ‘凤丹白’‘Feng Dan Bai’凤丹品种群
    Feng Dan cultivar group
    单瓣型
    Single type
    白色
    White
    ‘姚黄’ ‘Yao Huang’中原牡丹品种群
    Zhongyuan cultivar group
    皇冠型
    Crown type
    淡黄色
    Light yellow
    ‘High Noon’Lemoine系
    Lemoine strain
    荷花型
    Lotus type
    黄色
    Yellow
    ‘Dao Da Chen’日本牡丹品种群
    Japanese cultivar group
    菊花型
    Chrysanthemum type
    蓝紫色
    Blue-pruple
    下载: 导出CSV

    表  2  杂交结实统计

    Table  2.   Seed-set of different cross combinations

    杂交类型
    Hybrid type
    母本
    Female parent
    父本
    Male parent
    授粉花朵数
    Number of pollinated flower
    结籽数
    Total number of seed
    单花结籽数
    Seed number per flower
    组内近缘杂交
    Hybridization in Lactiflora group
    ‘朱砂判’‘Zhu Sha Pan’‘Kansas’301735.77
    ‘朱砂判’‘Zhu Sha Pan’‘Taff’362045.67
    ‘朱砂判’‘Zhu Sha Pan’‘Karl Rosenfield’271595.89
    组内远缘杂交Interspecific hybridization in sect. Paeonia‘朱砂判’‘Zhu Sha Pan’‘Garden Peace’21291.38
    ‘朱砂判’‘Zhu Sha Pan’‘Pink Teacup’22492.23
    ‘朱砂判’‘Zhu Sha Pan’‘Cream Delight’128790.62
    组间杂交
    Intersectional hybridization
    ‘朱砂判’‘Zhu Sha Pan’‘凤丹白’‘Feng Dan Bai’5000
    ‘朱砂判’‘Zhu Sha Pan’‘姚黄’‘Yao Huang’5000
    ‘朱砂判’‘Zhu Sha Pan’‘High Noon’5100
    ‘朱砂判’‘Zhu Sha Pan’‘Dao Da Chen’3390.27
    下载: 导出CSV

    表  3  亲本和杂交后代的核型参数(‘朱砂判’× ‘Cream Delight’)

    Table  3.   Chromosome parameters of parents and progenies ('Zhu Sha Pan'× 'Cream Delight')

    品种
    Cultivar
    相对长度
    Relative length/%
    臂比值
    Arm ratio
    最长染色体/最短染色体
    The longest chromosome/The shortest chromosome
    核不对称系数
    Asymmetry index/%
    着丝粒指数
    Centromere/%
    核型分类
    Karyotype
    核型公式
    Formula of karyotype
    ‘朱砂判’‘Zhu Sha Pan’16.40~25.021.28~5.001.5363.7116.67~43.782A2n=2x=10=6m+2sm+2st
    ‘Cream Delight’15.23~23.611.17~3.661.5559.9821.48~46.012A2n=4x=20=12m+4sm+4st
    ZC115.22~26.111.10~3.001.7259.1925.00~49.462A2n=2x=10=6m+2sm+2st
    ZC216.03~24.391.42~4.641.5262.1817.72~46.562A2n=2x=10=6m+2sm+2st
    ZC315.96~24.201.34~3.981.5261.9720.09~44.942A2n=2x=10=6m+2sm+2st
    ZC417.68~23.811.23~4.231.3561.6119.11~44.932A2n=3x=15=9m+3sm+3st
    ZC514.87~25.681.35~4.741.7362.6117.42~45.692A2n=3x=15=9m+3sm+3st
    ZC616.24~24.801.22~3.601.5362.4121.73~45.052A2n=3x=15=9m+3sm+3st
    ZC712.38~27.331.33~3.332.2162.7223.12~42.912B2n=3x=15=9m+3sm+3st
    注:m代表染色体臂比值为1.01~1.70;sm代表染色体臂比值为1.71~3.00;st代表染色体臂比值为3.01~7.00。Notes:metacentric (m):the arm ratio of each chromosome is from 1.01 to 1.70; submetacentric (sm):the arm ratio of each chromosome is from 1.71 to 3.00; subtelocentric (st):the arm ratio of each chromosomes is from 3.01 to 7.00.
    下载: 导出CSV

    表  4  SSR多态性引物

    Table  4.   SSR primers of Paeonia lactiflora

    引物(序列符号)
    Primer sequence(No.)
    序列(5′-3′)
    Primer sequence (5′-3′)
    重复基序
    Repeat motif
    目的片段
    Target fragment/bp
    退火温度Annealing temperature/℃
    44F  CACAATCAAAGTTTAAAGTGGTGATGT
    R CGTAGATGAGCACTTGTATTCTTCCT
    (AG)13A29358
    45F  GGAAAGGAGAAAGAGGACACTCA
    R  CACATCGGGATTTGTTAGCG
    (AC)20(TA)929255
    55F  CCAGTTATGGCGTCGTCTACTCTT
    R  GGAAAGGAAGGCAAGCTGATG
    (CT)1321357
    S3 (CL1109Contig1)F  CCCTCCCTTTCTTCACTTCC
    R  TTGAAACCCTAAACCCACCA
    (CT)822755
    S8 (CL17410Contig1)F  GAATCCACGAAAAATACGCA
    R  AGCGGGATGTAACCCTTCTT
    (AG)1225453
    S16 (CL25347Contig1)F  TTGAACAGCATTTGCACCTC
    R  ATGCGCTCTCTCTGTTTGGT
    (AT)824554
    S30(CL9771Contig1)F  TGAAAGGTGCACCATCAAAA
    R  ACGATCCCTCCTTTTCCATT
    (AG)924352
    下载: 导出CSV

    表  5  亲本和杂交后代经7对SSR多态性引物扩增结果

    Table  5.   Amplified results of hybrids and their parents by 7 SSR primers

    品种
    Cultivar
    引物Primer
    444555S3S8S16S30
    ‘朱砂判’‘Zhu Sha Pan’282, 286270, 286220, 224228, 238208255, 275243
    ‘Cream Delight’282, 296262, 270, 272, 286224, 226228, 238156, 208245, 255, 273, 275243, 247
    ZC1282270224, 226232, 238208255, 275243
    ZC2282, 286286, 288214, 224228, 238208245243
    ZC3282, 286286214, 224228, 238208245243, 247
    ZC4282, 294270224, 226228156, 208273, 275243
    ZC5282, 286270224, 226228156, 208273, 275243
    ZC6282, 286286220, 224228208255243
    ZC7282, 286262, 286220, 224228, 238208273, 275243
    下载: 导出CSV
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出版历程
  • 收稿日期:  2016-05-27
  • 修回日期:  2016-11-11
  • 刊出日期:  2017-04-01

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