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基于荧光SSR标记的白杨派种质资源遗传多样性研究

姚俊修 毛秀红 李善文 刘学良 吴德军

姚俊修, 毛秀红, 李善文, 刘学良, 吴德军. 基于荧光SSR标记的白杨派种质资源遗传多样性研究[J]. 北京林业大学学报, 2018, 40(6): 92-100. doi: 10.13332/j.1000-1522.20170429
引用本文: 姚俊修, 毛秀红, 李善文, 刘学良, 吴德军. 基于荧光SSR标记的白杨派种质资源遗传多样性研究[J]. 北京林业大学学报, 2018, 40(6): 92-100. doi: 10.13332/j.1000-1522.20170429
Yao Junxiu, Mao Xiuhong, Li Shanwen, Liu Xueliang, Wu Dejun. Genetic diversity of germplasm resources of Leuce based on SSR fluorescent marker[J]. Journal of Beijing Forestry University, 2018, 40(6): 92-100. doi: 10.13332/j.1000-1522.20170429
Citation: Yao Junxiu, Mao Xiuhong, Li Shanwen, Liu Xueliang, Wu Dejun. Genetic diversity of germplasm resources of Leuce based on SSR fluorescent marker[J]. Journal of Beijing Forestry University, 2018, 40(6): 92-100. doi: 10.13332/j.1000-1522.20170429

基于荧光SSR标记的白杨派种质资源遗传多样性研究

doi: 10.13332/j.1000-1522.20170429
基金项目: 

山东省重点研发计划项目 2017GNC11115

山东省农业良种工程项目 2012213

详细信息
    作者简介:

    姚俊修,博士,工程师。主要研究方向:林木遗传育种。Email:yjx95289528@163.com 地址: 250014 山东省济南市历下区文化东路42号山东省林业科学研究院

    责任作者:

    李善文,博士,研究员。主要研究方向:林木遗传育种。Email: lishanwen66@163.com 地址:同上

  • 中图分类号: S722.3

Genetic diversity of germplasm resources of Leuce based on SSR fluorescent marker

  • 摘要: 目的进行白杨派种质资源的遗传多样性研究,为毛白杨种质资源保存、评价与利用提供参考。方法对所收集的白杨派无性系采用毛细管电泳法进行荧光SSR-PCR产物检测,利用所得结果评价种源及无性系间的遗传变异和遗传多样性水平。结果16对SSR引物对272个无性系进行检测,共检测到106个等位基因,每位点的等位基因数为4~11个,平均等位基因数为6.625个;平均观测杂合度(Ho)值为0.561,平均期望杂合度(He)值为0.432,表明遗传多样性比较丰富;有9个位点的Ho/He大于1,表明杂合度比较高。对其中来自毛白杨6个集中分布区的234个无性系进行遗传变异分析,结果表明:2%的遗传变异来源于种源间,98%的遗传变异来自于种源内无性系间;由不同种源间的等位基因模型可知,山西、陕西、河南、山东和河北5个种源的无性系均有特有等位基因,杂交时可以适当选择不同种源的无性系作杂交亲本以拓宽杂交种的遗传基础,或种源间适当引种增加毛白杨种源间的遗传多样性。结论聚类分析和主坐标分析结果均表明,北京与河北种源亲缘关系最近,河南与陕西、山东与山西亲缘关系也较近,分别聚为3大类群,而相同种源的无性系并没有完全聚到一类,其原因可能是由于种源间相互引种或遗传变异造成的。研究结论为毛白杨种质资源保存和利用提供了理论依据。

     

  • 图  1  种源的等位基因模型

    NaF.等位基因频率;PA.特有等位基因数;CA.共有等位基因数(≤25%)。

    Figure  1.  Allelic patterns across provenances

    NaF, allele frequency; PA, private alleles; CA, common alleles.

    图  2  毛白杨种源和无性系的遗传变异

    Figure  2.  Genetic variations of provenances and clones of Populus tomentosa

    图  3  6个种源的主坐标分析

    Figure  3.  Main coordinate analysis of six provenances

    图  4  基于Nei's遗传距离的毛白杨不同种源UPGMA系统关系图

    Figure  4.  UPGMA clustering tree of Populus tomentosa provenances based on Nei's genetic distance

    图  5  基于16对SSR引物的272个白杨派无性系聚类图

    Figure  5.  Cluster diagram of 272 clones of Leuce based on 16 SSR primers

    表  1  不同种源取样数及无性系编号

    Table  1.   Number of samples from different provenances and clone No.

    来源
    Source
    样本数
    Sample number
    无性系编号
    Clone No.
    北京Beijing 25 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25
    河北Hebei Province 56 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81
    山东Shandong Province 19 82, 83, 221, 222, 232, 234, 235, 237, 242, 243, 244, 245, 246, 247, 248, 249, 251, 254, 255
    河南Henan Province 41 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 223, 225, 226, 227, 228, 229, 230, 257, 258, 259
    山西Shanxi Province 44 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 260
    陕西Shaanxi Province 49 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 185, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 238, 239, 261
    甘肃Gansu Province 5 204, 205, 206, 207, 208
    安徽Anhui Province 9 209, 210, 211, 212, 213, 214, 215, 216, 217
    江苏Jiangsu Province 4 218, 219, 220, 263
    白杨杂种White poplar hybrid
    (WPH)
    19 224, 231, 233, 236, 240, 241, 250, 252, 253, 256, 262, 264, 266, 267, 268, 269, 270, 271, 272
    新疆杨P. bolleana 1 265
    下载: 导出CSV

    表  2  引物序列信息

    Table  2.   Primer sequence information

    SSR编码
    SSR code
    重复单元
    Repeat motif
    SSR位置
    Position of SSR
    正向引物序列
    LP sequence
    反相引物序列
    RP sequence
    Ptr_1_SSR1 [CCT]5 61857~61870 AAAGCTTGTGTTCCACTTGT CAGATCTACCTCCTCCATCA
    Ptr_1_SSR2 [AAGA]6 98103~98127 TCTCAATTACATCCCAATCC GGTTGATTCACCAGCAGTAT
    Ptr_3_SSR13 [TA]10 12067876~12067894 AGTTGTTTGGGCTGTGTATC GTGCAATTCCCTGATTTAAG
    Ptr_7_SSR14 [ATT]6 1689228~1689246 TCCCACAAGCACTCTTAACT CCTTTGCAGCACAGTAGTAA
    Ptr_7_SSR15 [TG]12 1788775~1788798 CACTTGCTCTTACTCCTGCT CCAGGACAAATGCAATACTT
    Ptr_9_SSR3 [CGA]7 464617~464636 TCGAATTCTCCGATAGTGTT AATGCTTTCTTATGCTGCTC
    Ptr_10_SSR1 [GAC]9 364056~364082 CCAACAACAAGTACCCTCAT AGCTAGAGTCCTGTCTGCTG
    Ptr_11_SSR1 [TA]29 32355~32412 TGAATGAAATACATTGCTGC GCTATTATTGGATTTGCCTG
    Ptr_11_SSR8 [AT]30 3040852~3040911 AAATGGAGACTTGTGTGGAC GTGCAATAAAGCAGTGTGAA
    Ptr_13_SSR6 [GAA]8 732759~732781 TGGTACTCTCCTCTTGCCTA CCTTCAGTTTCTGCTTCATC
    Ptr_14_SSR7 [GCG]6 714903~714919 TGATTCTACTGGATCCAACC TATCCGATTCTCTAAGGCAA
    Ptr_14_SSR11 [TCC]8 1130007~1130029 CTCTCAGTTCTCTTGGATCG GAGGTCTCATTTGTTGAAGC
    Ptr_14_SSR12 [AT]13 1300753~1300777 TACTGGTGGTGCTCAATACA AAAGCAAACGCAGTAATAGC
    Ptr_16_SSR3 [AT]10 567670~567689 GGGATTTCACCACTTATTGA TTATATTTGTCTGGGAGCGT
    Ptr_18_SSR17 [AT]13 6997967~6997992 TTTCTGATGCTGTAGCTGTG ATCAGTATGCTTTGCCTGTT
    Ptr_19_SSR1 [TCA]8 110734~110756 ATTTCTTTCGCCTCATAACA CCCTCTTTGTGTGGAATTTA
    下载: 导出CSV

    表  3  16个位点272个白杨派样品SSR引物多态性分析

    Table  3.   Polymorphism analysis of 272 clones of Leuce at 16 SSR locus

    位点
    Locus
    样本量
    Sample number
    等位基因数
    Allele number
    有效等位基因数
    Effective number of allele
    Shannon信息指数
    Shannon information index
    观测杂合度
    Observed heterozygosity(Ho)
    期望杂合度
    Expected heterozygosity(He)
    多态信息指数
    Polymorphic information index
    Ptr-1-SSR2 272 5.000 2.236 0.898 0.952 0.553 0.388
    Ptr-7-SSR14 272 9.000 3.992 1.573 0.897 0.750 0.691
    Ptr-1-SSR1 272 5.000 1.840 0.837 0.287 0.456 0.423
    Ptr_9_SSR3 272 5.000 1.098 0.238 0.022 0.089 0.021
    Ptr_11_SSR1 272 7.000 2.604 1.134 0.923 0.616 0.494
    Ptr_10_SSR1 272 6.000 2.390 1.000 0.982 0.582 0.597
    Ptr_11_SSR8 272 5.000 1.038 0.113 0.018 0.036 0.004
    Ptr_14_SSR12 272 9.000 2.060 1.058 0.599 0.515 0.429
    Ptr_14_SSR7 272 5.000 2.054 1.033 0.537 0.513 0.461
    Ptr_16_SSR3 272 6.000 1.034 0.109 0.015 0.033 0.000
    Ptr_3_SSR13 272 7.000 1.463 0.583 0.279 0.316 0.224
    Ptr_14_SSR11 272 6.000 2.088 0.812 0.978 0.521 0.378
    Ptr_18_SSR17 272 11.000 3.103 1.446 0.482 0.678 0.594
    Ptr_7_SSR15 272 9.000 2.832 1.249 0.993 0.647 0.536
    Ptr_13_SSR6 272 7.000 2.343 1.012 0.985 0.573 0.576
    Ptr_19_SSR1 272 4.000 1.034 0.098 0.033 0.033 0.013
    均值Mean 272 6.625 2.075 0.825 0.561 0.432 0.364
    下载: 导出CSV

    表  4  16个位点234个毛白杨无性系的SSR多态性分析

    Table  4.   Polymorphism analysis of 234 clones of Populus tomentosa at 16 SSR locus

    位点
    Locus
    样本量
    Sample number(N)
    等位基因数
    Allele number(Na)
    有效等位基因数
    Effective number of allele(Ne)
    Shannon信息指数
    Shannon information index(I)
    观测杂合度
    Observed heterozygosity(Ho)
    期望杂合度
    Expected heterozygosity(He)
    Ptr-1-SSR2 234 3.000 2.034 0.736 0.996 0.508
    Ptr-7-SSR14 234 9.000 3.716 1.488 0.932 0.731
    Ptr-1-SSR1 234 3.000 1.882 0.824 0.303 0.469
    Ptr_9_SSR3 234 3.000 1.022 0.066 0.004 0.021
    Ptr_11_SSR1 234 4.000 2.378 0.961 0.966 0.579
    Ptr_10_SSR1 234 6.000 2.866 1.214 0.996 0.651
    Ptr_11_SSR8 234 2.000 1.004 0.015 0.004 0.004
    Ptr_14_SSR12 234 6.000 1.868 0.903 0.577 0.465
    Ptr_14_SSR7 234 5.000 1.985 0.965 0.534 0.496
    Ptr_16_SSR3 234 1.000 1.000 0.000 0.000 0.000
    Ptr_3_SSR13 234 2.000 1.347 0.426 0.295 0.257
    Ptr_14_SSR11 234 3.000 2.009 0.707 1.000 0.502
    Ptr_18_SSR17 234 8.000 2.783 1.275 0.470 0.641
    Ptr_7_SSR15 234 6.000 2.576 1.048 1.000 0.612
    Ptr_13_SSR6 234 6.000 2.811 1.160 0.996 0.644
    Ptr_19_SSR1 234 4.000 1.013 0.046 0.013 0.013
    平均Mean 234 4.438 2.018 0.740 0.568 0.412
    下载: 导出CSV

    表  5  6个种源毛白杨无性系遗传多样性分析

    Table  5.   Genetic diversity of 6 provenances of Populus tomentosa

    种源
    Provenance
    代码
    Code
    N Na Ne I Ho He
    北京Beijing BJ 25.000 2.375 1.895 0.630 0.593 0.391
    河北Hebei Province HB 56.000 3.125 1.912 0.672 0.578 0.393
    山东Shandong Province SD 18.000 2.750 1.845 0.631 0.524 0.361
    河南Henan Province HN 42.000 3.125 2.091 0.735 0.563 0.415
    山西Shanxi Province SX 44.000 3.750 1.912 0.706 0.544 0.391
    陕西Shaanxi Province SHX 49.000 3.250 2.071 0.747 0.585 0.423
    平均Mean 39.000 3.063 1.954 0.687 0.564 0.396
    下载: 导出CSV

    表  6  不同种源的特有等位基因和频率

    Table  6.   Private alleles and frequencies of different provenances

    种源
    Provenance
    位点
    Locus
    等位基因
    Allele
    特有等位基因频率
    Private allele and gene frequency
    河北Hebei Province Locus2 354 0.009
    山东Shandong Province Locus2 369 0.028
    Locus8 343 0.028
    河南Henan Province Locus6 235 0.012
    Locus7 294 0.012
    山西Shanxi Province Locus2 393 0.011
    Locus6 229 0.011
    Locus8 345 0.011
    Locus9 253 0.011
    Locus12 348 0.011
    Locus14 347 0.011
    Locus14 355 0.011
    Locus15 345 0.011
    Locus15 357 0.011
    Locus16 292 0.011
    陕西Shaanxi Province Locus4 269 0.02
    Locus13 193 0.01
    Locus14 359 0.02
    Locus16 280 0.01
    Locus16 281 0.01
    下载: 导出CSV

    表  7  6个种源间的Nei's遗传距离

    Table  7.   Nei's genetic distance among six provenances

    种源Provenance 北京
    Beijing
    河北
    Hebei Province
    山东
    Shandong Province
    河南
    Henan Province
    山西
    Shanxi Province
    陕西
    Shaanxi Province
    北京Beijing 0
    河北Hebei Province 0.004 0
    山东Shandong Province 0.032 0.02 0
    河南Henan Province 0.034 0.022 0.02 0
    山西Shanxi Province 0.012 0.004 0.01 0.014 0
    陕西Shaanxi Province 0.066 0.047 0.041 0.016 0.035 0
    下载: 导出CSV

    表  8  白杨派无性系聚类图各个分支的数量及其来源

    Table  8.   Quantity and source of Leuce clones in each branch of cluster diagram

    分类
    Classification
    北京
    Beijing
    河北
    HebeiProvince
    山东
    ShandongProvince
    河南
    HenanProvince
    山西
    ShanxiProvince
    陕西
    ShaanxiProvince
    甘肃
    GansuProvince
    安徽
    AnhuiProvince
    江苏
    JiangsuProvince
    新疆杨
    P. bolleana
    白杨杂种
    WPH
    总计
    Total
    1 12 21 4 11 12 10 1 2 2 0 0 75
    2 12 29 7 7 24 8 3 2 0 0 0 92
    3 0 0 0 0 1 0 0 0 1 1 11 14
    4 0 2 0 6 4 20 0 0 1 0 0 33
    5 1 4 8 17 3 11 1 5 0 0 8 58
    总计Total 25 56 19 41 44 49 5 9 4 1 19 272
    下载: 导出CSV
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  • 收稿日期:  2017-11-29
  • 修回日期:  2018-01-16
  • 刊出日期:  2018-06-01

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