番茄属种间杂种离体培养F1植株的细胞学分析

本文报道了栽培番茄（Lycopersico esculentum)“北京早红”等5个品种分别与野生型秘鲁番茄（L.peruvianum)PI128657中8号株系杂交,离体胚培养,得F1杂种植株,对花粉母细胞在减数分裂中染色体行为和终变期二价体交叉点的频率,以及亲和性程度等进行了.结果表明,6个亲本植株花粉母细胞减数分裂染色体的行为是正常的,中期I为12个二价体。其中环状二价体占多数,棒状二价体数较少,中期I没有单价体,后期I和II均正常,四分体阶段无微出现,但各亲本在终变期和中期I的环状二价体和棒状二价体的数有一定的差异,这可能与不同亲本基因型的亲和性程度和在遗传学上的不协调有关。5个组合的大多数F1杂种花粉母细胞减数分裂中染色体行为基本正常,12个二价体占多数,但染色体配对不稳定,有较多的单价体,染色单体桥。四分体阶段有微核,此外,在5个组合的F1杂种植株中,均出现双二倍体花粉母细胞,这些双二倍体花粉母细胞的染色体,在减数分裂中,也均,出现落后染色体和染色单体桥,以及较多的多价体,四分体阶段有微和不同类型的四分孢子群。     

Iris japonica×Iris confusa种间杂种的细胞遗传学研究

对Irisjaponica、I confusa和它们的人工种间杂种花粉母细胞减数分裂中期Ⅰ染色体配对行为、形态学和繁育特征进行了分析 ,结果表明 :(1)I japonica和I confusa的形态差异较小 ,杂种F1形态介于亲本之间 ;(2 )人工杂交比较容易 ,杂种种子能正常发育 ;杂种F1体细胞染色体数为 2n =30 ,减数分裂中期I染色体配对频率很高 ,为 13 9个二价体 ,构型为0 95Ⅰ + 5 95Ⅱ (棒形 ) + 7 95Ⅱ (环形 ) + 0 0 5Ⅲ + 0 2 7Ⅳ ,表明它们有相似的染色体组 ,亲缘关系很近 ;(3)杂种中有少量的三价体和四价体存在 ,可能是亲本染色体间发生了结构重排或是部分同源染色体配对 ;(4)杂种中大多数细胞存在 2个单价体 ,有的高达 8个单价体 ;花粉育性为 5 0 5 1% ,不能正常结实 ,表明I japonica和I confusa间存在生殖隔离 ,是独立的生物学物种 ;(5 )杂种F1减数分裂前期可观察到细胞融合 ,这可能是造成I japonica和I confusa不同居群出现多倍体和非整倍体的重要原因。     

东乡野生稻与栽培稻正反交种间杂种F_1的雄配子发生与发育

该文以东乡野生稻与栽培稻（美国光壳稻P1003）的正反交种间杂种F1（正交为光壳稻P1003×东乡野生稻;反交为东乡野生稻×光壳稻P1003）为试材,研究其各个减数分裂时期的染色体行为特征、染色体交换重组及雄配子发育特点。结果表明：正反交杂种F1花粉母细胞细胞核减数分裂的同步性较高,细胞质为连续型胞质分裂类型。在细胞核分裂的过程中,核仁在前期I到中期I逐渐消失。染色体在前期I到中期I逐渐收缩,变得更加清晰可见。在终变期和中期I,90.54%以上的花粉母细胞能形成12个二价体（含少数棒状二价体和8字型二价体）,部分花粉母细胞（正交9.46%,反交7.65%）出现少量的单价体、三价体和四价体。后期I观察到1.27%–1.35%的花粉母细胞含有1至数条落后染色体。最终有92.6%–94.8%的小孢子能发育成形态正常、染色能力强的成熟花粉粒。另外,在正交杂种F1的粗线期中存在2个核仁,而反交杂种F1及其双亲的粗线期只有1个核仁。这些研究结果可为作物品种改良、种质资源创新以及种间亲缘关系研究提供细胞学证据。     

Cucumis sativus × C.hystrix种间杂种的形态学和细胞学观察

以甜瓜属种间杂种F1(2n=19,华南型黄瓜"二早子"×Cucumis hystrix)为试材,对其形态学、细胞学和育性作了观察分析.结果表明该杂种无雌花,雄花不能正常开放,表现高度不育.该杂种F1长势瘦弱,与笔者以前报道的种间杂种F1(C. hystrix与华北型黄瓜"北京截头"正反交)相比,在育性和形态学性状上有明显差异.花粉母细胞减数分裂观察发现,杂种F1的终变期和中期Ⅰ主要以17条单价体(Ⅰ)和1个二价体(II)存在;整个花粉母细胞的减数分裂行为异常,经常可见染色体滞后和纺锤丝定向紊乱,形成多极染色体,末期II后形成多分体,以致不能发育成正常的花粉粒,导致杂种F1高度不育.     

八倍体小偃麦与天蓝偃麦草杂交F1染色体组构型

首次获得麦草8号、麦草9号、远中2号八倍体小偃麦与天蓝偃麦草的属间杂种,杂交当代结实率为31.49%,39.28%和10.41%。杂种F1表现为两亲的中间型,植株高大、繁茂,穗长20~30 cm,小穗数25~30个,多年生,抗寒,在哈尔滨冬季无覆盖条件下可安全过冬。对F1植株进行减数分裂行为观察,结果发现,染色体配对不正常,单价体频率高,出现多价体。杂种F1减数分裂中期I染色体配对构型分别为:9.5Ⅰ+16.98Ⅱ+0.27Ⅲ、13.6Ⅰ+14.01Ⅱ+0.87Ⅲ、11.2Ⅰ+16.8Ⅱ+0.08Ⅲ。二价体数变动在13~18间、单价体数变动在11~17间、多价体变动在0.08~0.87间,二价体多数是棒状二价体,推测两亲有一对部分同源关系染色体组,其余为非同源染色体组,但有的染色体间有部分同源关系。小麦5B染色体上Ph基因可能受到E组染色体的抑制。     

水稻亚种间杂种F1半不育小孢子发生的细胞学观察

利用石蜡切片和染色体压片法对水稻亚种间半不育杂种F1及其亲本的小孢子母细胞减数分裂过程进行细胞学观察.结果显示:亲本及杂种F1的花药壁发育正常,但部分F1的小孢子母细胞减数分裂异常,形成不均等的二分体和异常的四分体;其染色体在中期Ⅰ分散在赤道板两旁或远离赤道板,形成单价体;在后期Ⅰ和后期Ⅱ产生大量落后染色体或染色体桥.研究表明,部分花粉母细胞减数分裂中期和后期染色体行为异常可能是造成杂种F1花粉半不育的主要原因.     

Hystrix patula与Pseudoroegneria libanotica属间杂种的细胞学研究

为研究猬草Hystrixpatula的染色体组组成,进行了H.patula与Pseudoroegnerialibanotica的人工杂交,获得杂种F1,观察了亲本和杂种F1花粉母细胞减数分裂染色体配对行为。杂种F1染色体配对较高,84%的细胞形成7个或7个以上二价体,其构型为6.08Ⅰ+7.48Ⅱ,C-值为0.69。结果表明,H.patula含有St染色体组。     

东乡野生稻与栽培稻正反交种间杂种F1的雄配子发生与发育

该文以东乡野生稻与栽培稻(美国光壳稻P1003)的正反交种间杂种F₁(正交为光壳稻P1003×东乡野生稻; 反交为东乡野生稻×光壳稻P1003)为试材, 研究其各个减数分裂时期的染色体行为特征、染色体交换重组及雄配子发育特点。结果表明: 正反交杂种F₁花粉母细胞细胞核减数分裂的同步性较高, 细胞质为连续型胞质分裂类型。在细胞核分裂的过程中, 核仁在前期I到中期I逐渐消失。染色体在前期I到中期I逐渐收缩, 变得更加清晰可见。在终变期和中期I, 90.54%以上的花粉母细胞能形成12个二价体(含少数棒状二价体和8字型二价体), 部分花粉母细胞(正交9.46%, 反交7.65%)出现少量的单价体、三价体和四价体。后期I观察到1.27%–1.35%的花粉母细胞含有1至数条落后染色体。最终有92.6%–94.8%的小孢子能发育成形态正常、染色能力强的成熟花粉粒。另外, 在正交杂种F₁的粗线期中存在2个核仁, 而反交杂种F₁及其双亲的粗线期只有1个核仁。这些研究结果可为作物品种改良、种质资源创新以及种间亲缘关系研究提供细胞学证据。     

FISH分析栽培高粱×拟高粱、甜高粱×拟高粱的染色体行为

采用荧光原位杂交技术对45S rDNA在栽培高粱×拟高粱、甜高粱×拟高粱F1的有丝分裂和减数分裂染色体进行定位研究。在有丝分裂中期染色体上2个杂种分别检测到2个杂交信号, 在减数分裂粗线期、终变期、中期Ⅰ染色体上45S rDNA位于一个二价体上, 说明这两个杂种携带45S rDNA的染色体为同源染色体。根据45S rDNA位点随细胞减数分裂过程的位置变化, 表明这两个杂种染色体配对行为正常, 平均构型为2n=2x=20(10Ⅱ), 证明45S rDNA可作为染色体的一个识别指标间接地观察细胞减数分裂过程染色体的变化行为。     

大鹅观草与蛇河披碱草属间杂种的细胞遗传学研究

为探讨大鹅观草(Roegneria grandis,2n=4x=28)的染色体组组成,为其正确的分类处理提供细胞学依据。该研究通过人工远缘杂交,成功获得3株大鹅观草与蛇河披碱草(Elymus wawawaiensis,2n=4x=28)属间杂种F1植株。杂种植株形态介于两亲本之间,不育。亲本及杂种经I2-IK溶液染色后进行花粉育性检测,结果显示Roegneria grandis和Elymus wawawaiensis的花粉可育,育性高达94.6%和90.5%;杂种F1不育。花粉母细胞减数分裂中期I染色体配对结果显示,亲本花粉母细胞配对正常,均形成14个二价体,以环状二价体为主,Roegneria grandis有频率很低(0.04/细胞)的单价体出现;杂种F1平均每个花粉母细胞形成6.46个二价体,变化范围为5~8;在观察的83个花粉母细胞中,有35.2%的花粉母细胞形成了7个二价体,形成6个二价体的细胞占42.59%,较少细胞形成8个二价体;平均每个细胞形成14.66个单价体,变化范围为10~18;平均每细胞观察到0.14个三价体;杂种花粉母细胞染色体构型为14.66 I+6.46 II+0.14 III;平均每细胞交叉数为9.83,C值为0.35。结果表明:(1)R.grandis与Elymus wawawaiensis有一组染色体组同源的St染色体组,另外一组染色体组不是St或者H染色体组,Roegneria grandis的染色体组组成不是St Stg;(2)较低频率的三价体(平均0.14个/细胞),可能是由于R.grandis的St和Y染色体组间具有一定的同源性,也可能是染色体易位等原因导致,对于Y染色体组的起源还需深入地研究;(3)在不同地理来源的披碱草属和鹅观草属物种中St染色体组同源性不同,R.grandis与来自于北美的Elymus lanceolatus与E.wawawaiensis的St染色体组较与分布于亚洲的E.sibiricus和E.caninus的St染色体组同源性反而更高,其原因还需要进一步地研究。     

Cytological studies on meiosis and male gametophyte development in autotetraploid cucumber

With improved staining and chromosome preparation techniques, meiosis of pollen mother cells (PMCs) and male gametophyte development in autotetraploid cucumber (Cucumis sativus L.) was studied to understand the correlation between chromosomes behaviour and fertility. Various chromosome configurations, e.g. multivalent, quadrivalents, trivalents, bivalents and univalents were observed in most PMCs at metaphase I. Lagging chromosomes were frequently observed at anaphase in both meiotic divisions. In addition, chromosomes segregations were not synchronous and equal in some PMCs during anaphase II and telophase II. Dyads, triads, tetrads with micronuclei and polyads were observed at tetrad stage, and the frequencies of normal tetrad with four microcytes were only 55.4 %. The frequency of abnormal behaviour in each stage of meiosis was counted, and the average value was 37.2 %. The normal meiotic process could be accomplished to form the microspore tetrads via simultaneous cytokinesis. Most microspores could develop into fertile gametophytes with 2 cells and 3 germ pores through the following stages: single-nucleus early stage, single-nucleus late stage and 2-celled stage. The frequency of abnormalities was low during the process of male gametophyte development. The germination rate of pollen grains was 46.9 %. These results suggested that abnormal meiosis in PMCs was the reason for low pollen fertility in the autotetraploid cucumber.     

Meiotic behavior of pollen mother cells in relation to ploidy level of somatic hybrids between Solanum tuberosum and S. chacoense

Potato somatic hybrids obtained by protoplast fusion between Solanum tuberosum (4x) and Solanum chacoense (2x) were investigated for genome stability and meiotic behavior associated with the pollen viability in order to elucidate the mechanism influencing the fertility of the somatic hybrids. The ploidy level detections conducted in 2004 and 2007 demonstrated that 68 out of 108 somatic hybrids had their ploidy level changed to be uniform and euploidy after successive in vitro subcultures, which mainly occurred in octaploids, aneuploids, and mixoploids, while 74% hexaploids were still stable in their genome dosage in 2007. Different types of abnormal meiotic behavior were observed during the development of pollen mother cells (PMCs) including the formation of univalents, multivalents, laggard chromosomes, and chromosomal bridges, as well as triads and polyads. A higher proportion of abnormal meiosis seemed to be accompanied with a genome dosage higher than the hexaploids expected in this study. A significant positive correlation between defective PMCs and the number of small pollen grains and negative correlation between number of small pollen grains and pollen viability strongly suggested that abnormal meiosis could be a causal factor influencing the fertility of the somatic hybrids. The hexaploids with stable genome dosage and a certain level of fertility will have great potential in a potato breeding program.     

部分菊属植物及其种间杂种减数分裂异常现象观察

对不同倍数性菊属植物及其部分种间杂种的减数分裂异常现象进行观察统计,并分析其形成机制以及在菊属系统演化中的作用。结果表明,菊属减数分裂异常现象包括分裂不同步、二价体提前解离、二价体互锁、染色体桥、落后染色体等。减数分裂不同步现象普遍存在于菊属植物减数分裂过程。二倍体的菊花脑、甘菊、异色菊的部分二价体在终变期提前解离为单价体。菊花脑及其部分杂种中观察到了互锁二价体。四倍体菊花脑、南京野菊、‘黄英’、‘滁菊’在AI和AII都出现了染色体桥,毛华菊有1.5%的PMC在AI出现染色体桥。四倍体菊花脑AI、AII期出现落后染色体的频率分别为10.6%和7.3%;毛华菊AI期出现落后染色体的频率为4.4%;栽培菊‘黄英’和‘滁菊’在AI、AII期出现落后染色体的频率高于毛华菊。杂种出现染色体桥及落后染色体的频率普遍高于亲本。倒位以及由其引起的各种染色体结构变异可能在菊属系统演化过程中起着重要作用。     

萝卜与甘蓝属间杂种基因组原位杂交分析

用基因组原位杂交方法（Genomic in situ hybridization, 简称GISH）研究了萝卜( Raphanus sativus,2n=18,RR)和甘蓝（Brassica oleracea , 2n=18, CC）属间杂种F1减数分裂过程。结果表明杂种体细胞染色体组成为RC,2n=18,但花粉母细胞有三种不同类型：1. RC,2n=18, 终变期染色体平均配对构型为14.87Ⅰ+1.20Ⅱ+0.04Ⅲ+0.06Ⅳ, 染色体配对主要发生在萝卜和甘蓝染色体之间, 后期Ⅰ9条萝卜染色体主要以5/4和6/3的分离比移向两极, 所形成配子的染色体数目和组成均不平衡,配子败育; 2. RRCC,4n=36, 终变期染色体形成18个二价体,后期Ⅰ染色体均衡分离,形成RC不减数配子;3. RRCC缺体,4n=30－34, 少数萝卜染色体丢失,形成的配子具有全套的甘蓝染色体和部分萝卜染色体。     

白菜型油菜与蓝花子杂交的初步研究

通过胚胎培养,成功地获得了白菜型油菜（Ｂｒａｓｉｃａｃａｍｐｅｓｔｒｉｓ）与蓝花子（ＲａｐｈａｎｕｓｓａｔｉｖｕｓＬｖａｒｒａｐｈａｎｉｓｔｒｏｉｄｅｓＭａｋｉｎｏ）的属间杂种。该杂种具有两种类型;一种为大花类型,一种为小花类型。对它们进行花粉母细胞减数分裂的观察结果表明：小花类型为未加倍的杂种ＭＩ,存在１９个未配对染色体,大花类型为加倍或部分加倍杂种,加倍类型ＭＩ,１９个二价体排列在赤道板上;部分加倍类型ＡＩ,具有１０－１０－９的染色体组分割现象。大花类型具有可育性;它能够产生很多ｎ＝１９及ｎ＝９、ｎ＝１０的正常配子。染色体组分割能够产生倍半二倍体,它能用来研究染色体的功能和开展染色体工程。     

Formation and function of phragmoplast during successive cytokinesis stages in higher plant meiosis

Cytoskeletal rearrangements were studied during meiotic telophase in a number of monocotyledonous plant species. Wild type and abnormal meiosis (in wide cereal hybrids, meiotic mutants and allolines) was analyzed. It was found that central spindle fibers that move centrifugally, along with newly-formed MTs, are the basis of phragmoplast formation and function in PMCs of monocotyledonous plant species with successive cytokinesis stages. A model for centrifugal movement of the meiotic phragmoplast is proposed; this model is a modification of the corresponding process during B-anaphase.     

Cytological investigations of the interspecific hybrids of Nicotiana tabacum L. x N. glauca Grah

Interspecific amphihaploid and amphidiploid hybrids between Nicotiana glauca Grah. (2n = 24) and N. tabacum L. (2n = 48) cultivars BY 103 and K 326 were analysed. F1 amphihaploids (2n = 36) were viable and completely self- and cross-sterile, and mostly univalents were present during meiosis (with pairing range from 0 to 5). In some meiocytes, meiotic irregularities were observed, such as sporadic chromatin bridges and formation of restitution nuclei. The resultant F1 hybrids were easily converted to amphidiploids (2n = 72) via colchicine treatment of seedlings. The number of univalents and the frequency of PMCs containing unpaired chromosomes indicated that amphidiploids N. tabacum cv. BY 103 or K 326 x N. glauca represented quite a high pairing category. However, they were male sterile because pollen mother cells were arrested at the tetrad stage. The termination of development of PMCs, and consequently male sterility, are very rare in this kind of tobacco hybrids.     

MS32-regulated timing of callose degradation during microsporogenesisin Arabidopsis is associated with the accumulation of stacked rough ER in tapetal cells

  In the male sterile32(ms32)mutant in Arabidopsis thaliana, pollen development is affected during meiosis of pollen mother cells (PMCs). In normal wild-type (WT) anthers, callose is deposited around PMCs before and during meiosis, and after meiosis the tetrads have a complete callose wall. In ms32, PMCs showed initial signs of some callose deposition before meiosis, but it was degraded soon after, as was part of the cellulosic wall around the PMCs. The early dissolution of callose in ms32 was associated with the occurrence of extensive stacks of rough ER (RER) in tapetal cells. The stacks of RER were also observed in the WT tapetum, but at a later stage, i.e., after the tetrads were formed and when callose is normally broken down for release of microspores. Based on these observations it is suggested that: (1) callose degradation around developing microspores is linked to the formation of RER in tapetal cells, which presumably synthesize and/or secrete callase into the anther locule, and (2) mutation in MS32 disrupts the timing of these events. Received: 27 April 1999 / Revision accepted: 21 June 1999     

The role of microtubular cytoskeleton and callose walls in the cytomixis process in tobacco (Nicotiana tabacum L.) pollen mother cells

We have studied the microtubule cytoskeleton structure and callose walls deposition in the course of meiosis at the cytomictic and normal tobacco (N. tabacum L.) PMCs. It was ascertained that microtubule cytoskeleton did not play an evident part in the process of cytomixis. Increased cytomixis frequency probably is determined by irregular callose walls deposition. The possible reasons of nuclear material passage between tobacco PMCs at the cellular level are discussed.     

Accessory phragmoplast corrects abnormal cytokinesis in wheat x rye hybrids

The compensation for phragmoplast dysfunction in the male meiosis of F1 wheat × rye hybrids was described. In pollen mother cells (PMCs), he transition from central spindle fibers (forming a solid bundle) to phragmoplast (hollow cylinder) was blocked. This blockage suppresses the centrifugal movement of the phragmoplast and cell-plate formation. As a result, cells become binucleate. Sometimes, two nuclei fuse and form one restitution nucleus. In PMCs of the wheat × rye F1 hybrid D-144 gp 06 year (T. aestivum n. 93-60 t 9 × S. cereale n. Saratovskaya 7) with this phenotype, an additional phragmoplast is formed at the late telophase. This occurs by a common mechanism for the development of the immobile phragmoplast in the meiosis in bicotyledons; new phragmoplasts arise as a result of microtubule polymerization starting from the spindle poles. The accessory phragmoplast facilitates a new cell plate assembly and achievement of cytokinesis.