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山西省吉县刺槐无性系种质遗传多样性的EST-SSR分析

李秀宇 郭琪 董黎 孙宇涵 牛东升 刘佳平 王红生 李云

李秀宇, 郭琪, 董黎, 孙宇涵, 牛东升, 刘佳平, 王红生, 李云. 山西省吉县刺槐无性系种质遗传多样性的EST-SSR分析[J]. 北京林业大学学报, 2019, 41(7): 39-48. doi: 10.13332/j.1000-1522.20190108
引用本文: 李秀宇, 郭琪, 董黎, 孙宇涵, 牛东升, 刘佳平, 王红生, 李云. 山西省吉县刺槐无性系种质遗传多样性的EST-SSR分析[J]. 北京林业大学学报, 2019, 41(7): 39-48. doi: 10.13332/j.1000-1522.20190108
Li Xiuyu, Guo Qi, Dong Li, Sun Yuhan, Niu Dongsheng, Liu Jiaping, Wang Hongsheng, Li Yun. EST-SSR analysis of genetic diversity of Robinia pseudoacacia clones in Jixian County, Shanxi Province of northern China[J]. Journal of Beijing Forestry University, 2019, 41(7): 39-48. doi: 10.13332/j.1000-1522.20190108
Citation: Li Xiuyu, Guo Qi, Dong Li, Sun Yuhan, Niu Dongsheng, Liu Jiaping, Wang Hongsheng, Li Yun. EST-SSR analysis of genetic diversity of Robinia pseudoacacia clones in Jixian County, Shanxi Province of northern China[J]. Journal of Beijing Forestry University, 2019, 41(7): 39-48. doi: 10.13332/j.1000-1522.20190108

山西省吉县刺槐无性系种质遗传多样性的EST-SSR分析

doi: 10.13332/j.1000-1522.20190108
基金项目: 国家自然科学基金项目(31570677),国家重点研发课题(2017YFD0600503),北京林业大学重大科研成果培育项目(2017CGP007),国家林业局科技发展中心项目(2016007)
详细信息
    作者简介:

    李秀宇。主要研究方向:林木生物技术与良种繁育。Email:lixiuyu@bjfu.edu.cn 地址:100083 北京市海淀区清华东路35号北京林业大学生物科学与技术学院

    责任作者:

    李云,教授,博士生导师。主要研究方向:林木遗传育种。Email:yunli@bjfu.edu.cn 地址:同上

EST-SSR analysis of genetic diversity of Robinia pseudoacacia clones in Jixian County, Shanxi Province of northern China

  • 摘要: 目的了解各对EST-SSR引物检出的等位简单序列重复片段数量,解析吉县良种基地刺槐无性系种质的多态性状况与种质间亲缘关系,为引物利用和刺槐种质遗传管理提供参考。方法利用课题组开发的45对EST-SSR引物进行PCR扩增,使用毛细管电泳仪对产物进行检测,用Popgene 1.32版软件对群体遗传参数进行评估,利用NTsys 2.10e进行聚类分析。结果45对EST-SSR引物中,除了引物RP-24均表现出良好的多态性。44对引物共检测到等位重复序列298个,每对引物检测到2 ~ 15个,平均6.77个。群体观测杂合度(Ho)的变化范围为0.022 7 ~ 0.842 1,平均为0.412 6。期望杂合度(He)的变化范围为0.088 7 ~ 0.822 3,平均为0.484 0。引物多态性信息量(PIC)变化范围为0.085 2 ~ 0.796 6,平均为0.444 4。结论该基地的刺槐无性系种质具有较高的遗传多样性。由聚类结果可知,在遗传距离为0.81时,可将96份刺槐无性系种质聚成5类。聚类图在分子水平上显示了刺槐无性系种质间的亲缘关系,研究结果对于利用这些引物用于刺槐的种质评价、资源管理和辅助育种有积极的参考价值。

     

  • 图  1  基于EST-SSR数据的96份刺槐资源聚类图

    Figure  1.  Dendrogram of 96 Robinia pseudoacacia resources based on EST-SSR data

    表  1  试验所用EST-SSR引物信息

    Table  1.   Information of EST-SSR primers used in the experiment

    位点 Locus 引物序列 Primer sequence (5′−3′) 重复基元 Repeat motif 大小 Size/bp 退火温度 Ta/℃
    Rp-01 F: TGCAGAAAGAGAAAGCAGAGG (TGTGAA)4 140 58
    R: CCGAACCCTTTCTGGTTAGTC
    Rp-02 F: GCTGCGTTTAATTTTGTCAGG (GAAT)4 170 58
    R: TCAATCCATCAAAGAGGAAACA
    Rp-03 F: GTGAGAAGTGGTTAGGGTTTT (CTC)7 186 55
    R: TCAAGATCACCAACGTACAA
    Rp-04 F: CTCGTGATGATGGTGTTGATG (AATGGT)4 146 58
    R: AATGGTCCAAACAACACGAAG
    Rp-05 F: CCTTGCACATTTATCCCAGAA (TCTGGC)3 158 57
    R: CGACCTCGATCTTTTCTTGTG
    Rp-06 F: TGGACAAAACATCATCGTGTG (TGAGTT)4 147 59
    R: CTCTCTTCTTTCTGCCCCTCA
    Rp-07 F: TTTTTCTCCCAACGAAACAAA (CT)10 144 56
    R: TGATGTGTTGTACGGAGGTGA
    Rp-08 F: TCAGGTGCATAAGCTCATTACTTC (AAAAT)4 152 56
    R: GGTTGTCAGATGAAATGCACA
    Rp-09 F: CGTTTAGAAGCTGAGGCAGAA (CTTT)5 153 58
    R: TGAGATATCTTAGTGCAGGAGCA
    Rp-10 F: GGCATGTGGCTATGAAGATGT (CCTTT)4 154 57
    R: TCAGTGGGACTTGGTTTCTTG
    Rp-11 F: GAAGCTATCACCGCAAATGAA (AG)10 150 57
    R: GTCGAAGTGCGTCCTAGATCA
    Rp-12 F: AAGAGTCATCACGGAGACCAA (AGCAGA)4 150 56
    R: GGAGTCCAATTAAGTGCGAGA
    Rp-13 F: CATTTCCGATTTCCAATTCCT (CTCTTC)4 151 56
    R: GCCGAGGACTCGGTAGAAGT
    Rp-14 F: TTAGCACGAACCTGGTTATGG (TGCAAC)4 151 56
    R: CACTTCATTTGGTTCCTTGAGA
    Rp-15 F: TTAACTAATGCGGCGAGAAGA (TCAC)5 119 56
    R: GAGAGGAAGTGTGCGAAACAA
    Rp-16 F: TATGAGACAGTGTTGGTTGGT (TTCAGT)4 175 56
    R: CGTGCCAGAAGAGTATAACAG
    Rp-17 F: GTAAGTCTGCAAAGAAGACCA (AACCA)4 150 56
    R: GCTTTTCACCTATCAACTCAA
    Rp-18 F: GGATGAACTTTGGCAATCCTT (GGTCAG)4 158 55
    R: AATTTGTTGGGAATGCTGTTG
    Rp-19 F: CAGGAGTGGCAGCATTAGTGT (AGGCTG)4 123 56
    R: CACAACAAGCACATTTTGCAC
    Rp-20 F: TTTCTTGGCTTGCTTTTGCTA (GCAGCT)3 145 56
    R: TCTTGGATACGCAAGGTTGTC
    Rp-21 F: TATGATCACGTCCCCTAATGC (CCA)7 146 57
    R: AAGTGGAAAGAAATGGGATGG
    Rp-22 F: GGTAAGGTGAAGGAGGTGGAG (AGGGTT)4 150 56
    R: AGCTTGGTCTCCTAGGTCGTC
    Rp-23 F: GGAGGAGCAACCATCTGTGTA (AGAAGT)4 146 56
    R: CTCCCTCTTCATCCTCACCTC
    Rp-24 F: TGCACATATTTGCCTGGTTTA (AATA)4 160 56
    R: AAAATGAGCATGACACAACCA
    Rp-25 F: CGGCAACAAGTTGAGAAGAAC (AAAG)5 139 56
    R: GGCTCACAAACCAACCTATGA
    Rp-26 F: GCTGCAAGCAAAGGATCTTAC (ATGATA)4 139 56
    R: CCTCATCATCCTCGTCATCAT
    Rp-27 F: TGGACAAAACATCATCGTGTG (TGAGTT)4 147 57
    R: CTCTCTTCTTTCTGCCCCTCA
    Rp-28 F: CTTGGTCTAGAAAGTCCTGCT (CAG)7 151 56
    R: GGTCATCAAGGTTAGTTGGAT
    Rp-29 F: CCTGATGATCAAAACGACGAC (GATC)4 148 56
    R: GGAGGTGACCCCTCTTATCCT
    Rp-30 F: TTGAACCAAAACTGGAAGAGC (GCT)8 151 56
    R: GCACCGTACAGTTACCCTATCC
    Rp-31 F: GACCCCATTTTTCTCAAGGAC (ATT)7 140 56
    R: TTGGATAAGTCGGTGAAGGTG
    Rp-32 F: CCACGTGGTTCTTCAAACATT (GTG)7 163 57
    R: CAACAACAACCCACAAACACA
    Rp-33 F: CAAACAGTCTCATGGAAATGGA (ATC)7 141 56
    R: GGGTTGGTATTGTTGGGAAAT
    Rp-34 F: AGGATATTAGCCAAGTCCATC (TGGTGA)4 164 57
    R: AGTAACCATCACCACAATCAC
    Rp-35 F: TCAGACGTGGTAGAGCAGTGTT (CACAC)4 152 58
    R: ATTTGTTTTTGGGGGAGATTG
    Rp-36 F: CGTTTCAGCCATTGATTTTGT (GAATC)5 141 57
    R: GATCATCACCGTCCACCTTC
    Rp-37 F: TGTCGTCATTTTATTTTACCC (GAACGA)4 152 56
    R: CTCACCCTTTTTATTTCCATT
    Rp-38 F: TCCATTCCCTGGTTTCTTCTT (TC)10 150 56
    R: AGCACAATTTCCTCAGTGCAG
    Rp-39 F: TTAAAGAATGTTCCGTTCAGA (AAGAGG)3 152 56
    R: GAGAAGATAGCCTCCTAGCTG
    Rp-40 F: TCATTGGACATCCCTCCATAA (TAA)8 139 56
    R: GGCTCGACATGGTTGATTTT
    Rp-41 F: AACTCACCCAATTGCACACTC (CCA)7 143 56
    R: GAGCAAGAGCTAAAGCAGCAA
    Rp-42 F: CTTCGCAATCCTCACTCTTTG (AATC)4 169 55
    R: CTTACCCAGAAGCCAACAATG
    Rp-43 F: CAAAGCAGAGAGAATGTATGG (CAAAAT)4 155 57
    R: ATCCCTTGCTCCTTGTAATAG
    Rp-44 F: TATCTGGGAGAATCGAGAGCA (ATCA)5 145 57
    R: CCACCATGGTTGTCCTTCTAA
    Rp-45 F: GGGTTGAGGAAGAGAGGAGAA (TTC)7 156 57
    R: AAAAATCGAATCGTGTTGGTG
    下载: 导出CSV

    表  2  44对引物扩增结果及多态性信息

    Table  2.   Amplification results and polymorphic information of 44 pairs of primers

    引物 Primer 观测杂合度 Ho 期望杂合度 He Nei氏指数 Nei 等位重复序列 Na 有效等位重复序列 Ne Shannon信息指数 I 引物多态性信息量 PIC
    RP-01 0.623 7 0.629 4 0.626 0 6 2.674 0 1.152 8 0.557 1
    RP-02 0.106 7 0.125 3 0.124 4 2 1.142 1 0.244 9 0.116 8
    RP-03 0.641 3 0.628 1 0.624 6 11 2.664 1 1.329 8 0.571 7
    RP-04 0.781 6 0.790 8 0.786 2 8 4.678 0 1.638 4 0.752 4
    RP-05 0.540 2 0.673 0 0.669 2 15 3.022 8 1.680 0 0.652 5
    RP-06 0.734 0 0.738 4 0.734 4 7 3.765 6 1.445 5 0.690 5
    RP-07 0.842 1 0.816 8 0.811 5 11 5.303 9 1.892 0 0.788 2
    RP-08 0.505 4 0.726 6 0.722 7 7 3.606 8 1.426 1 0.679 1
    RP-09 0.648 9 0.660 8 0.657 3 15 2.918 1 1.529 9 0.615 7
    RP-10 0.433 0.422 9 0.420 6 5 1.725 8 0.798 4 0.375 7
    RP-11 0.719 1 0.822 3 0.817 7 14 5.485 5 2.018 3 0.796 6
    RP-12 0.750 0 0.776 8 0.772 7 10 4.400 1 1.646 2 0.737 7
    RP-13 0.315 8 0.362 2 0.360 3 9 1.563 2 0.878 1 0.350 0
    RP-14 0.395 6 0.708 7 0.704 8 10 3.387 6 1.556 5 0.671 7
    RP-15 0.416 7 0.371 6 0.369 6 8 1.586 4 0.802 4 0.345 2
    RP-16 0.162 8 0.173 2 0.172 2 4 1.208 0 0.374 9 0.163 6
    RP-17 0.250 0 0.247 7 0.246 4 7 1.327 0 0.572 3 0.236 1
    RP-18 0.434 2 0.599 3 0.595 4 7 2.471 5 1.085 8 0.512 3
    RP-19 0.446 8 0.610 6 0.607 4 7 2.547 1 1.118 6 0.528 9
    RP-20 0.046 0 0.088 7 0.088 2 3 1.096 7 0.203 8 0.085 2
    RP-21 0.246 8 0.280 6 0.278 8 6 1.386 6 0.624 5 0.266 4
    RP-22 0.602 2 0.595 9 0.592 7 5 2.455 0 1.141 4 0.546 7
    RP-23 0.341 2 0.502 1 0.499 1 6 1.996 4 0.957 2 0.446 5
    RP-25 0.104 2 0.139 2 0.138 5 5 1.160 7 0.345 2 0.134 9
    RP-26 0.212 8 0.214 0 0.212 8 6 1.270 4 0.472 0 0.201 0
    RP-27 0.118 3 0.112 5 0.111 9 3 1.126 0 0.242 6 0.106 6
    RP-28 0.651 7 0.719 5 0.715 4 7 3.514 2 1.436 0 0.671 6
    RP-29 0.022 7 0.107 8 0.107 2 2 1.120 0 0.218 1 0.101 4
    RP-30 0.268 8 0.416 6 0.414 3 5 1.707 4 0.775 8 0.367 3
    RP-31 0.376 6 0.561 2 0.557 6 10 2.260 4 1.269 3 0.532 9
    RP-32 0.511 9 0.635 9 0.632 1 7 2.718 0 1.246 2 0.578 3
    RP-33 0.557 9 0.444 1 0.441 7 4 1.791 2 0.696 4 0.357 3
    RP-34 0.500 0 0.523 5 0.520 6 8 2.086 0 1.106 7 0.485 3
    RP-35 0.375 0 0.565 1 0.561 9 5 2.282 7 0.987 2 0.486 1
    RP-36 0.512 2 0.661 5 0.657 4 4 2.919 0 1.167 6 0.590 9
    RP-37 0.612 9 0.665 1 0.661 5 6 2.954 4 1.353 5 0.628 0
    RP-38 0.556 8 0.774 9 0.770 5 15 4.357 9 1.895 0 0.741 3
    RP-39 0.392 4 0.592 3 0.588 5 5 2.430 3 1.021 3 0.507 8
    RP-40 0.178 9 0.182 0 0.181 0 3 1.221 0 0.345 7 0.166 4
    RP-41 0.315 8 0.331 3 0.329 5 6 1.491 5 0.706 5 0.311 7
    RP-42 0.281 7 0.459 5 0.456 3 2 1.839 1 0.648 7 0.352 2
    RP-43 0.221 1 0.305 3 0.303 7 4 1.436 1 0.613 7 0.285 1
    RP-44 0.081 4 0.133 3 0.132 6 4 1.152 8 0.317 1 0.129 0
    RP-45 0.315 2 0.398 4 0.396 2 4 1.656 2 0.661 0 0.333 4
    总计 Total 298 104.907 6
    均值 Mean 0.412 6 0.484 0 0.481 2 6.772 7 2.384 3 0.991 9 0.444 4
    下载: 导出CSV
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出版历程
  • 收稿日期:  2019-03-28
  • 修回日期:  2019-05-26
  • 网络出版日期:  2019-07-05
  • 刊出日期:  2019-07-01

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