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‘正午’牡丹核型分析及减数分裂的染色体行为观察

钟原 杜明杰 刘羽心 成仿云

钟原, 杜明杰, 刘羽心, 成仿云. ‘正午’牡丹核型分析及减数分裂的染色体行为观察[J]. 北京林业大学学报, 2019, 41(10): 68-73. doi: 10.13332/j.1000-1522.20180381
引用本文: 钟原, 杜明杰, 刘羽心, 成仿云. ‘正午’牡丹核型分析及减数分裂的染色体行为观察[J]. 北京林业大学学报, 2019, 41(10): 68-73. doi: 10.13332/j.1000-1522.20180381
Zhong Yuan, Du Mingjie, Liu Yuxin, Cheng Fangyun. Karyotype analysis and meiotic chromosome behavior observation on Paeonia × lemoinei ‘High Noon’[J]. Journal of Beijing Forestry University, 2019, 41(10): 68-73. doi: 10.13332/j.1000-1522.20180381
Citation: Zhong Yuan, Du Mingjie, Liu Yuxin, Cheng Fangyun. Karyotype analysis and meiotic chromosome behavior observation on Paeonia × lemoinei ‘High Noon’[J]. Journal of Beijing Forestry University, 2019, 41(10): 68-73. doi: 10.13332/j.1000-1522.20180381

‘正午’牡丹核型分析及减数分裂的染色体行为观察

doi: 10.13332/j.1000-1522.20180381
基金项目: 北京林业大学青年教师科学研究中长期项目(2015ZCQ-YL-03)
详细信息
    作者简介:

    钟原,博士,副教授。主要研究方向:牡丹育种与栽培。Email:zhongyuanbjfu@126.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:

    成仿云,教授,博士生导师。主要研究方向:牡丹育种与栽培。Email:chengfy8@263.net  地址:同上

  • 中图分类号: S722

Karyotype analysis and meiotic chromosome behavior observation on Paeonia × lemoinei ‘High Noon’

  • 摘要: 目的芍药属牡丹组革质花盘亚组与肉质花盘亚组间的远缘杂交是现代牡丹育种的重要方向之一。然而,亚组间杂种普遍高度不育,很难继续用于杂交育种。‘正午’牡丹是一个观赏性好、适应性强的亚组间杂交品种,虽然其通常高度不育,但仍被用作亲本培育出一些优异的杂交后代,表现出一定的育性。研究其减数分裂的染色体行为,可为揭示其极低育性的形成机制提供重要的信息。方法本研究以‘正午’幼嫩花蕾中的雌蕊为材料,进行体细胞染色体核型分析;以花药为材料进行花粉母细胞减数分裂观察。结果‘正午’牡丹为二倍体,核型公式为2n = 2x = 10 = 7m + 1sm + 2st(1SAT)。其花粉母细胞可通过正常的减数分裂形成四分体及小孢子,减数第一次分裂中约有70%的花粉母细胞发生染色体行为异常,包括单价体及多价体、染色体桥、断片、落后染色体、不等分裂等,其中染色体桥出现频率最高;减数第二次分裂中,染色体行为异常率同样高达70%,常见的异常类型包括纺锤体定位异常、不同步分裂、染色体桥、断片、落后染色体等,最后形成二分体、三分体、微核或特小的额外小孢子。结论‘正午’减数分裂存在大量异常可能与其高度杂合的核型有关。普遍存在的单价体及多价体引起的不等分裂和染色体桥及断片造成的染色体片段缺失等染色体行为异常可能是导致‘正午’牡丹高度不育的重要原因。同时,仍有一小部分花粉母细胞能够顺利完成减数分裂,形成小孢子。其中一部分通过二分体或三分体形成未减数小孢子,表明其具有用于培育多倍体牡丹的潜力。

     

  • 图  1  ‘正午’牡丹中期染色体及核型

    Figure  1.  Metaphase chromosomes and karyotype of Paeonia × lemoinei ‘High Noon’

    图  2  ‘正午’牡丹花粉母细胞的正常减数分裂过程

    a:中期I,5个整齐排列的棒状二价体;b、c:后期I;d:末期I;e:中期II;f:后期II;g:末期II;h:四分体时期。 a, metaphase I with 5 rod bivalents orderly arranged; b,c, anaphase I; d, telophase I; e, metaphase II; f, anaphase II; g, telophase II; h, tetrad.

    Figure  2.  Regular meiosis of pollen mother cells in Paeonia × lemoinei ‘High Noon’

    图  3  减数第一次分裂中期的异常

    a ~ e:4个二价体 + 2个单价体(a、e:4个棒状二价体 + 2个单价体,b ~ d:3个棒状二价体 + 1个环形二价体 + 2个单价体);f ~ j:3个二价体 + 4个单价体;k、l:2个二价体 + 6个单价体(k:棒状二价体 + 环形二价体 + 6个单价体,l:2个棒状二价体 + 6个单价体);m:1个二价体 + 8个单价体;n:1个二价体 + 1个三价体 + 5个单价体;o:2个二价体 + 1个三价体 + 3个单价体;p:3个二价体 + 1个三价体 + 1个单价体;q:1个二价体 + 1个七价体 + 1个单价体;r:1个二价体 + 1个五价体 + 3个单价体 + 1个断片;s:1个五价体 + 5个单价体;t:1个六价体 + 4个单价体。a−e, 4 bivalents + 2 univalents (a, e, 4 rod bivalents + 2 univalents, b−d, 3 rod bivalents + 1 ring bivalent + 2 univalents); f−j, 3 bivalents + 4 univalents; k−l, 2 bivalents + 6 univalents (k, rod bivalent + ring bivalent + 6 univalents; l, 2 bivalents + 6 univalents); m, 1 bivalent + 8 univalents; n, 1 bivalent + 1 trivalent + 5 univalents; o, 2 bivalents + 1 trivalent + 3 univalents; p, 3 bivalents + 1 trivalent + 1 univalent; q, 1 bivalent + 1 heptavalent + 1 univalent; r, 1 bivalent + 1 quinquevalent + 3 univalents + 1 fragment; s, 1 quinquevalent + 5 univalents; t, 1 hexavalent + 4 univalents.

    Figure  3.  Meiotic abnormalities in metaphase I

    图  4  减数第一次分裂后期到末期的异常

    a ~ c:有染色体桥,无断片;d ~ g:有染色体桥及1 ~ 6个断片;h、i:染色体双桥及断片;j ~ l:染色体桥和落后染色体;m:末期染色体桥及断片;n ~ s:无染色体桥,有落后染色体及断片;t:染色体不等分离;u ~ x:不等分离、染色体桥、落后染色体及断片同时存在。a−c, chromosome bridge with no fragment; d−g, chromosome bridge with 1−6 fragments; h−i, chromosome double bridge with fragments; j−l, chromosome bridge with lagging chromosome; m, chromosome bridge with fragment in telophase I; n−s, lagging chromosome and fragment with no chromosome bridge; t, unequal segregation of chromosomes; u−x, unequal segregation of chromosomes with chromosome bridge, lagging chromosome and fragment.

    Figure  4.  Meiotic abnormalities in anaphase I and telophase I

    图  5  减数第二次分裂的异常

    a ~ f:中期II的不同纺锤体定位(a:直线纺锤体,b:垂直纺锤体,c、d:三极纺锤体,e:融合纺锤体,f:平行纺锤体);g ~ n:减II后期的异常(g:染色体桥,h:对角线桥,i、j:三极纺锤体后期形成的2组减数染色体及1组未减数染色体,k:融合纺锤体后期形成的2组未减数染色体,l:4组染色体及额外染色体,m:后期II的不同步性,形成2组减数染色体及1组未减数染色体,n:后期II的对角线桥及染色体断片);o ~ t:末期II的异常(o、p、s:2个减数核和1个未减数核,p:3个核及染色体桥、断片,q:2个未减数核,r:4个核及2个微核,t:末期对角线桥);u:三分体;v:三分体和微核;w:二分体;x:2个正常小孢子、1个具微核的小孢子和1个特小的小孢子。a−f, aspects of meiotic spindles at metaphase II (a, linear spindle, b, perpendicular spindle, c−d, tripolar spindle, e, fused spindle, f, parallel spindle); g−n, meiotic abnormalities in anaphase II (g, chromosome bridge, h, chromosome diagonal bridge, i−j, 2 groups of reduced chromosomes and 1 group of unreduced chromosomes come from the tripolar spindles, k, 2 groups of unreduced chromosomes come from the fused spindles, l, 4 groups of chromosomes with extra chromosomes, m, 2 groups of reduced chromosomes and 1 group of unreduced chromosomes because of the asynchronized anaphase II, n, chromosome diagonal bridge and fragment in anaphase II); o−t, meiotic abnormalities in telophase II (o, p, s, 2 reduced nucleuses and 1unreduced nucleus, p, 3 nucleuses with chromosome bridge and fragment, q, 2 unreduced nucleuses, r, 4 nucleuses with 2 micronucleuses, t, diagonal bridge in telophase II); u, triad; v, triad with micronucleus; w, dyad; x, 2 regular microspores, 1 microspore with micronucleus and 1 extra small microspore.

    Figure  5.  Meiotic abnormalities in the second meiotic division

    表  1  ‘正午’牡丹核型数据

    Table  1.   Karyotype parameters of Paeonia × lemoinei ‘High Noon’

    染色体序号
    Chromosome No.
    相对长度
    Relative length/%
    臂比
    Arm ratio
    染色体类型
    Chromosome type
    1 11.77 ± 0.53 1.40 ± 0.05 m
    2 11.48 ± 0.53 1.17 ± 0.09 m
    3 10.94 ± 0.52 1.16 ± 0.08 m
    4 10.65 ± 0.23 1.52 ± 0.13 m
    5 10.25 ± 0.34 1.39 ± 0.10 m
    6 10.09 ± 0.24 1.15 ± 0.11 m
    7 9.59 ± 0.29 1.23 ± 0.21 m
    8 9.37 ± 0.47 1.71 ± 0.24 sm
    9 8.27 ± 0.59 4.39 ± 0.37 st
    10 7.58 ± 0.42 3.53 ± 0.17 st*
    注:*随体染色体,随体长度未计算在染色体长度内。Note: * represents sat-chromosome, the length of satellite was not included.
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  • 收稿日期:  2018-11-26
  • 修回日期:  2019-05-13
  • 网络出版日期:  2019-09-17
  • 刊出日期:  2019-10-01

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