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杨树腐烂病菌表观特征及遗传多样性分析

张俊娥 范鑫磊 梁英梅 田呈明

张俊娥, 范鑫磊, 梁英梅, 田呈明. 杨树腐烂病菌表观特征及遗传多样性分析[J]. 北京林业大学学报, 2017, 39(7): 76-86. doi: 10.13332/j.1000-1522.20160257
引用本文: 张俊娥, 范鑫磊, 梁英梅, 田呈明. 杨树腐烂病菌表观特征及遗传多样性分析[J]. 北京林业大学学报, 2017, 39(7): 76-86. doi: 10.13332/j.1000-1522.20160257
ZHANG Jun-e, FAN Xin-lei, LIANG Ying-mei, TIAN Cheng-ming. Analysis of appearance features and genetic diversity of Cytospora chrysosperma causing poplar canker[J]. Journal of Beijing Forestry University, 2017, 39(7): 76-86. doi: 10.13332/j.1000-1522.20160257
Citation: ZHANG Jun-e, FAN Xin-lei, LIANG Ying-mei, TIAN Cheng-ming. Analysis of appearance features and genetic diversity of Cytospora chrysosperma causing poplar canker[J]. Journal of Beijing Forestry University, 2017, 39(7): 76-86. doi: 10.13332/j.1000-1522.20160257

杨树腐烂病菌表观特征及遗传多样性分析

doi: 10.13332/j.1000-1522.20160257
基金项目: 

林业公益性行业科研专项 201204501

北京市支持中央高校共建项目 2050205

详细信息
    作者简介:

    张俊娥。主要研究方向:林木病理学。Email:1419031699@qq.com  地址:100083  北京市海淀区清华东路35号北京林业大学林学院

    责任作者:

    田呈明,教授,博士生导师。主要研究方向:林木病理学。Email:chengmt@bjfu.edu.cn  地址:同上

  • 中图分类号: S763.11

Analysis of appearance features and genetic diversity of Cytospora chrysosperma causing poplar canker

  • 摘要: 为探究杨树腐烂病菌的表观特征、遗传多样性以及它们与不同寄主杨树派别和病原菌地理来源的关系,本文收集来自我国14个省份的106份杨树腐烂病标本,纯培养观察记录菌落生长状况,分析菌株培养形态多样性,并从中选取背景信息差异较大的菌株34份,采用SRAP和ISSR两种标记方法分析其遗传多样性。结果显示:杨树腐烂病菌同一菌株在不同条件下的培养性状稳定;不同菌株的培养形态差异明显,且各类别间菌株的培养性状与寄主植物、地理来源没有明显相关性。利用获得多态性良好的12对SRAP引物和20条ISSR引物分析得出群体的Nei's多样性指数为0.31(SRAP)、0.28(ISSR),各菌株间存在较大的遗传差异;并从寄主角度分析其群体遗传结构,得出群体总的遗传多样性为0.27,群体内的遗传多样性为0.23,可以解释总体多样性的87%,遗传分化系数(GST)为0.13,遗传变异主要存在于群体内。从地理角度分析得出,总的群体遗传多样性为0.28,群体内的遗传多样性为0.13,可以解释总体多样性的48%,GST为0.52,来自群体内和群体间的多样性差异相当,而地理群体间的迁移对总体的遗传分化产生了较大的影响,各群体的遗传多样性水平表现为从西到东依次降低的趋势。

     

  • 图  1  不同培养条件下菌株性状稳定性比较

    Figure  1.  Stability comparison of strain characters under different culture conditions

    图  2  各类培养形态比对

    Figure  2.  Comparison in typical culture morphology

    图  3  杨树腐烂病菌培养形态聚类图

    Figure  3.  Clustering map of culture morphology in C. chrysosperma

    图  4  杨树腐烂病菌群体聚类图

    Figure  4.  Clustering map of C. chrysosperma

    表  1  菌株信息

    Table  1.   Information of strains

    菌株Strain寄主Host采集地Source
    C6941*箭杆杨Populus nigra var. thevestina北京海淀Haidian, Beijing
    C80322*毛白杨P. tomentosa北京海淀Haidian, Beijing
    C80323*毛白杨P. tomentosa北京海淀Haidian, Beijing
    C80327*北京杨P.× beijingensis北京海淀Haidian, Beijing
    C81310*欧美杨P.× canadensis陕西杨凌Yangling, Shaanxi
    C81311*毛白杨P. tomentosa陕西武功Wugong, Shaanxi
    C81312陕林3号杨P. deltides cv.‘Lux’ × P. deltoides陕西杨凌Yangling, Shaanxi
    C81313陕林3号杨P. deltides cv.‘Lux’ × P. deltoides陕西杨凌Yangling, Shaanxi
    C83121*小叶杨P. simonii内蒙古呼和浩特Hohhot, Inner Mongolia
    C83601毛白杨P. tomentosa四川汶川Wenchuan, Sichuan
    C83604杨树Populus sp.四川泸定Luding, Sichuan
    C85442*Ⅰ-69杨P. deltides cv.‘Lux’Ⅰ-69陕西榆林Yulin, Shaanxi
    C85447青杨P. cathayana陕西眉县Meixian County, Shaanxi
    C87640*Ⅰ-72杨P.× euramericana cv. ‘San Martino’江苏句容Jurong, Jiangsu
    G-YS-13*青杨P. cathayana甘肃临潭Lintan, Gansu
    G-YS-14*新疆杨P. alba. var. pyramidalis甘肃漳县Zhangxian County, Gansu
    G-YS-5-C杨树Populus sp.甘肃甘南Gannan, Gansu
    G-YS-3*青杨P. cathayana甘肃甘南Gannan, Gansu
    G-YS-9-1*小叶杨P. simonii甘肃甘南Gannan, Gansu
    JT-YS-2杨树Populus sp.吉林通化Tonghua, Jilin
    NW-1杨树Populus sp.内蒙古乌兰浩特Ulanhot, Inner Mongolia
    N-YS-10*小叶杨P. simonii宁夏永宁Yongning, Ningxia
    N-YS-11杨树Populus sp.宁夏永宁Yongning, Ningxia
    N-YS-13*小叶杨P. simonii宁夏永宁Yongning, Ningxia
    N-YS-14*小叶杨P. simonii宁夏永宁Yongning, Ningxia
    N-YS-15*新疆杨P. alba var. pyramidalis宁夏吴忠Wuzhong, Ningxia
    N-YS-4小叶杨P. simonii宁夏青铜峡Qingtongxia, Ningxia
    N-YS-7小叶杨P. simonii宁夏青铜峡Qingtongxia, Ningxia
    N-YS-8*小叶杨P. simonii宁夏中卫Zhongwei, Ningxia
    Q-YS-12新疆杨P. alba var. pyramidalis青海平安Ping'an, Qinghai
    Q-YS-13*青杨P. cathayana青海平安Ping'an, Qinghai
    Q-YS-14*新疆杨P. alba var. pyramidalis青海平安Ping'an, Qinghai
    Q-YS-16*新疆杨P. alba var. pyramidalis青海平安Ping'an, Qinghai
    Q-YS-19新疆杨P. alba var. pyramidalis青海互助Huzhu, Qinghai
    Q-YS-20*青杨P. cathayana青海互助Huzhu, Qinghai
    Q-YS-21*新疆杨P. alba var. pyramidalis青海互助Huzhu, Qinghai
    Q-YS-22*新疆杨P. alba var. pyramidalis青海互助Huzhu, Qinghai
    Q-YS-5*新疆杨P. alba var. pyramidalis青海乐都Ledu, Qinghai
    Q-YS-8*青杨P. cathayana青海乐都Ledu, Qinghai
    Q-YS-9青杨P. cathayana青海乐都Ledu, Qinghai
    XZ-YS-3杨树Populus sp.西藏乃东Naidong, Tibet
    XZ-YS-4杨树Populus sp.西藏乃东Naidong, Tibet
    XZ-YS-5杨树Populus sp.西藏乃东Naidong, Tibet
    XZ-YS-6杨树Populus sp.西藏乃东Naidong, Tibet
    XZ-YS-7银白杨P. alba西藏乃东Naidong, Tibet
    XZ-YS-8银白杨P. alba西藏乃东Naidong, Tibet
    XZ-YS-9银白杨P. alba西藏乃东Naidong, Tibet
    XZ-YS-9-V银白杨P. alba西藏乃东Naidong, Tibet
    ZK-110-1*杨树Populus sp.甘肃会宁Huining, Gansu
    ZK-148杨树Populus sp.北京通州Tongzhou, Beijing
    ZK-149*毛白杨P. tomentosa北京大兴Daxing, Beijing
    ZK-153-2杨树Populus sp.北京大兴Daxing, Beijing
    ZK-154杨树Populus sp.北京大兴Daxing, Beijing
    G-YS-9-2*小叶杨P. simonii甘肃甘南Gannan, Gansu
    HBPQ-1杨树Populus sp.河北平泉Pingquan, Hebei
    HBPQ-2杨树Populus sp.河北平泉Pingquan, Hebei
    HQ-15杨树Populus sp.黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HQ-18毛白杨P. tomentosa黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HQ-24杨树Populus sp.黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HQ-27杨树Populus sp.黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HQ-31杨树Populus sp.黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HQ-33杨树Populus sp.黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HQ-34大青杨P. ussuriensis黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HQ-35大青杨P. ussuriensis黑龙江齐齐哈尔Qiqihar, Heilongjiang
    HT-10大青杨P. ussuriensis黑龙江通北Tongbei, Heilongjiang
    HT-6大青杨P. ussuriensis黑龙江通北Tongbei, Heilongjiang
    HT-7大青杨P. ussuriensis黑龙江通北Tongbei, Heilongjiang
    HT-9杨树Populus sp.黑龙江通北Tongbei, Heilongjiang
    JA-5杨树Populus sp.西藏加查Gyaca, Tibet
    JAA-6杨树Populus sp.西藏加查Gyaca, Tibet
    JC-8杨树Populus sp.西藏加查Gyaca, Tibet
    JT-YS-1杨树Populus sp.吉林通化Tonghua, Jilin
    S-YS-3杨树Populus sp.陕西华县Huaxian County, Shaanxi
    S-YS-7毛白杨P. tomentosa陕西华县Huaxian County, Shaanxi
    S-YS-8毛白杨P. tomentosa陕西周至Zhouzhi, Shaanxi
    S-YS-9杨树Populus sp.陕西周至Zhouzhi, Shaanxi
    X-YS-14银灰杨P. canescens新疆轮台Luntai, Xinjiang
    X-YS-154银灰杨P. canescens新疆和田Hetian, Xinjiang
    X-YS-20毛白杨P. tomentosa新疆轮台Luntai, Xinjiang
    X-YS-294新疆杨P. alba var. pyramidalis新疆伊犁Ili, Xinjiang
    X-YS-383新疆杨P. alba var. pyramidalis新疆喀什Kashgar, Xinjiang
    X-YS-409新疆杨P. alba var. pyramidalis新疆阿克苏Aksu, Xinjiang
    X-YS-413银灰杨P. canescens新疆阿克苏Aksu, Xinjiang
    X-YS-419新疆杨P. alba var. pyramidalis新疆阿克苏Aksu, Xinjiang
    X-YS-433新疆杨P. alba var. pyramidalis新疆阿克苏Aksu, Xinjiang
    X-YS-78新疆杨P. alba var. pyramidalis.新疆和静Hejing, Xinjiang
    XZ-YS-1杨树Populus sp.西藏乃东Naidong, Tibet
    XZ-YS-10杨树Populus sp.西藏加查Gyaca, Tibet
    XZ-YS-11杨树Populus sp.西藏加查Gyaca, Tibet
    XZ-YS-12银白杨P. alba西藏加查Gyaca, Tibet
    XZ-YS-13银白杨P. alba西藏加查Gyaca, Tibet
    XZ-YS-14银白杨P. alba西藏加查Gyaca, Tibet
    XZ-YS-2杨树Populus sp.西藏乃东Naidong, Tibet
    ZK-157*毛白杨P. tomentosa北京大兴Daxing, Beijing
    ZK-158*欧美杨P.×canadensis北京大兴Daxing, Beijing
    ZK-163杨树Populus sp.北京大兴Daxing, Beijing
    ZK-182毛白杨P. tomentosa北京通州Tongzhou, Beijing
    ZK-183白杨树Populus sp.北京朝阳Chaoyang, Beijing
    ZK-188杨树Populus sp.北京通州Tongzhou, Beijing
    ZK-190欧美杨P.×canadensis北京通州Tongzhou, Beijing
    ZK-192欧美杨P.×canadensis北京通州Tongzhou, Beijing
    ZK-201杨树Populus sp.北京大兴Daxing, Beijing
    ZK-203杨树Populus sp.北京大兴Daxing, Beijing
    ZK-26-2*山杨P. davidiana北京门头沟Mentougou, Beijing
    ZK-59*新疆杨P. alba var. pyramidalis辽宁沈阳Shenyang, Liaoning
    ZK-60*新疆杨P. alba var. pyramidalis辽宁沈阳Shenyang, Liaoning
    注:*表示用于遗传多样性分析的菌株。Note: * means strain for genetic diversity analysis.
    下载: 导出CSV

    表  2  培养特征量化描述

    Table  2.   Quantitative description of culture characteristics

    特征赋值
    Assignment
    培养特征Culture characteristics
    色素
    Pigment
    生长速率
    Growth rate
    菌落质地
    Colony texture
    表面形态
    Surface morphology
    黏液
    Mucus
    子实体数量
    Fruiting quantity
    子实体形态
    Fruiting morphology
    1淡黄色
    Canary yellow

    Slow
    紧致
    Compact
    辐射
    Radiation

    No

    No

    No
    2黄棕色
    Yellow brown
    中等
    Medium
    疏松
    Loose
    非辐射
    Non radiation

    Yes

    Less

    Small
    3黑绿色
    Blackish green

    Fast
    极疏松
    Very loose
    同心圆
    Concentric circle

    More
    中等
    Medium
    4
    Big
    下载: 导出CSV

    表  3  SRAP标记引物序列

    Table  3.   Primer sequences of SRAP

    正向引物(5′-3′)
    Forward primer(5′-3′)
    反向引物(5′-3′)
    Reversed primer(5′-3′)
    1   5′-TGAGTCCAAACCGGATA-3′a   5′-GACTGCGTACGAATTAAT-3′
    2   5′-TGAGTCCAAACCGGAGC-3′b   5′-GACTGCGTACGAATTTGC-3′
    3   5′-TGAGTCCAAACCGGAAT-3′c   5′-GACTGCGTACGAATTGAC-3′
    4   5′-TGAGTCCAAACCGGACC-3′d   5′-GACTGCGTACGAATTTGA-3′
    5   5′-TGAGTCCAAACCGGAAG-3′e   5′-GACTGCGTACGAATTAAC-3′
    f   5′-GACTGCGTACGAATTGCA-3′
    下载: 导出CSV

    表  4  ISSR标记引物序列

    Table  4.   Primer sequences of ISSR

    编号
    No.
    引物序列(5′-3′)
    Primer sequence(5′-3′)
    IQ-1CGAATTCGACGACGACGA
    IQ-2CCAAACCACCACCACCACCA
    IJ-1AGAGAGAGAGAGAGAGG
    IJ-3AGAGAGAGAGAGAGAGTC
    IJ-4GAGAGAGAGAGAGAGATC
    IJ-5GAGAGAGAGAGAGAGATG
    IJ-7GGGTGGGGTGGGGTG
    IJ-8ATAAGAGAGAGAGAGAG
    IJ-9AGATGTGTGTGTGTGTG
    IL-1GGAGAGGAGAGGAGA
    IL-3CACGAGAGAGAGAGAGA
    IL-5CTCTCTCTCTCTCTCTT
    IL-7ACAACACACACACACAC
    ID-1AAAGTGTGTGTGTGTGT
    ID-2ATGATGATGATGATG
    ID-3ATGATGATGATGATGATG
    ID-6AGAAGAAGAAGAAGAAGAAGA
    ID-7GTCGTCGTCGTCGTCGTC
    ID-8GTGCGTGCGTGCGTGC
    ID-9CTCTCTCTCTCTCTCTG
    下载: 导出CSV

    表  5  杨树腐烂病菌群体遗传结构分布

    Table  5.   Population genetic structure of C. chrysosperma

    项目Item方法MethodHTHSGSTNm
    寄主角度Host aspectISSR0.253 40.216 20.147 12.899 0
    SRAP0.296 30.264 70.106 54.196 0
    ISSR & SRAP0.269 80.234 70.130 13.344 2
    地理Geography aspectISSR0.260 80.125 10.520 50.460 7
    SRAP0.309 40.147 80.522 40.457 1
    ISSR & SRAP0.279 40.133 70.521 30.459 1
    下载: 导出CSV
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  • 收稿日期:  2016-08-11
  • 修回日期:  2017-02-06
  • 刊出日期:  2017-07-01

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