中国春双端二体中端体的配对研究

21个中国春双端二体与中国春品种杂交,观察了杂种F_1减数分裂中期Ⅰ端体与相应染色体的配对。结果表明:14个组合的中期Ⅰ具有1个异形三价体的PMC的百分数超过90%;有3个组合(6A、5B和6D)分别为89.87%、83.56%和85.71%;2个组合(4A和1B)低于80%。在4A、1B和5B的3个组合中,具有1个异形二价体和1个单价端体的PMC超过15%。在4A、1B和4D的组合中,还有一定频率的PMC具有2个单价端体。用21个组合中端体配对频率计算的二价体频率与中国春品种中期Ⅰ构型频率基本一致。这些结果表明,染色体臂作为端体时和它作为双臂染色体的一部分时,是以同样的方便程度配对的。     

粘型小麦雄性不育系减数分裂特征及育性恢复研究

调查了粘型1B／1R和非1B／1R小麦雄性不育系,保持系及其F2的花粉母细胞减数分裂中期Ⅰ染色体联会情况、后期Ⅰ出现落后染色体的细胞频率以及末期Ⅱ含有微核的四分体的频率,结果表明：（1）粘果山羊细胞质对1B／1R型不育系减数分裂染色体配对水平具有特异性降低作用;（2）粘型1B／1R不育系减数分裂中期Ⅰ出现单价体细胞频率与后期Ⅰ出现落后染色体细胞的频率呈正相关,也与含微核的四分体频率呈正相关,而对应保持系则没有相关性;（3）粘果山羊草细胞质对非1B／1R不育系减数分裂过程影响不大,5个1B／1R不育系减数分裂过程中,3个时期染色体行为变异率的差异是特定的1B／1R核型与粘果山羊草细胞质互作的结果;（4）粘型1B／1R不育系杂交R2单株减数分裂3个时期染色体行为变异率与其恢复度成反比,这类不育系减数分裂中染色体行为不同步是其恢复不高且变异较大的一重要原因。     

粘型小麦不育系育性恢复性的细胞遗传学研究

利用5个粘性不育系分别与中国春的二体,1B重双端体及1B°1D^Ⅳ缺四体杂交,调查其F1的育性及其F1减数分裂中期Ⅰ的染色体配对情况和后期Ⅰ出现落后染色体的细胞频率。结果表明：（1）5个不育系与中国春二体杂交F1的自交结实率存在明显差异。中国春的1B重双端体使5个不育系的自交结实率大幅度下降,中国春的1B°1DⅣ缺四体使各不育系表现全不育;（2）5个不育系和3个父本杂交F1减数分裂中期Ⅰ出现单价体细胞的频率与其后期Ⅰ出现落后染色体细胞的频率不存在直接相关;（3）5个不育系与中国春二体的杂交F1自交结实率与其减数分裂中期Ⅰ出现单价体细胞的频率呈正相关。相关系数为0.9695**,与后期Ⅰ出现落后染色体细胞的频率不直接相关。     

同源四倍体矮牵牛花粉母细胞减数分裂观察

以同源四倍体矮牵牛06P-12为材料,采用常规压片法对花粉母细胞减数分裂过程及染色体行为进行了观察研究,以探明同源四倍体矮牵生育性降低的细胞学原因.结果显示:花粉母细胞减数分裂过程与二倍体基本相同但有其特殊性,主要表现在:终变期染色体的构型复杂,有四价体、三价体和单价体;中期Ⅰ和中期Ⅱ有赤道板外染色体;后期Ⅰ和后期Ⅱ出现落后染色体、丢失染色体、染色体桥及不均等分裂的现象;四分体时期出现一分体、二分休、三分体以及含微核的异常三分体、四分体、多分体.花粉母细胞减数分裂过程中正常细胞平均达78.6%,异常细胞频率平均为21.4%.研究表明,同源四倍体矮牵生育性降低的细胞学原因是减数分裂过程中染色体行为异常.     

水稻配子体、孢子体细胞质雄性不育系线粒体蛋白质的分析

利用SDS-聚丙烯酰胺凝胶电泳方法分析了水稻配子体细胞质雄性不育系粤泰A、保持系粤泰B、F_1代泰优2号、恢复系胜优2号和孢子体细胞质雄性不育系马协A、保持系马协B、F_1代马协63、恢复系明恢63及另一种孢子体细胞质雄性不育系珍汕97A、保持系珍汕97B、F_1代汕优63、恢复系明恢63黄化苗的线粒体蛋白质。结果表明,粤泰A、B、F_1、恢复系之间出现6条多肽带的差异,马协A、B、F_1、恢复系之间出现4条多肽带的差异,珍汕97A、B、F_1、恢复系之间出现2条多肽带的差异。     

小麦ph1b突变体与小黑麦杂交的细胞遗传学研究

本文利用普通小麦品系"中国春"(对照)、中国春ph1b突变体分别与八倍体小黑麦、六倍体小黑麦杂交,杂种F1的减数分裂前期Ⅰ染色体行为表现异常,中期Ⅰ出现较多的单价体、棒状二价体和多价体,在后期和末期出现落后染色体、染色体片断和微核。原因是ph1b基因的存在造成染色体联会机制紊乱,致使一些部分同源染色体配对并发生互换,有可能在以后的世代产生染色体易位与基因重组。     

几类异质小麦雄性不育系育性恢复性的细胞遗传学研究

系统调查了4类异质(粘果、易变、偏凸、二角山羊草细胞质)1BL/lRS、非1BL/1RS小麦雄性不育系与其恢复系杂种F减数分裂中期Ⅰ出现单价体细胞频率,以及后期Ⅰ出现落后染色体和染色体桥细胞频率,并对中、后期染色体变异率与杂种F自交结实率进行了相关分析.结果表明(1)1BL/1RS型杂种在中期Ⅰ、后期Ⅰ染色体变异率要明显高于非1BL/1RS杂种;(2)4类异源细胞质在非1BL/1RS杂种中有着明显提高单价体细胞频率的作用;(3)在1BL/1RS杂种中,1B@1BL/1RS杂合核型染色体联会松弛,对单价体频率的影响远大于异源细胞质的影响;(4)1BL/1RS型杂种自交结实率与中期出现单价体细胞频率不直接相关,而与后期出现落后染色体和染色体桥细胞的频率呈高度负相关;(5)非1BL/1RS型杂种在减数分裂中、后期染色体行为相对稳定,易恢复且恢复度高,很有实际利用价值.     

四倍体鲫鲤、三倍体湘云鲫染色体减数分裂观察

用精巢细胞直接制片法观察了异源四倍体鲫鲤、三倍体湘云鲫和二倍体红鲫、湘江野鲤精母细胞染色体第一次减数分裂中期配对情况 ;作为对照 ,观察了上述四种鱼肾细胞的有丝分裂中期染色体。在精母细胞第一次减数分裂中 ,异源四倍体鲫鲤同源染色体两两配对 ,形成 10 0个二价体 ,没有观察到单价体、三价体和四价体 ;三倍体湘云鲫精母细胞形成 5 0个二价体和 5 0个单价体 ;红鲫和湘江野鲤精母细胞分别形成 5 0个二价体。肾细胞检测表明异源四倍体的染色体数目为 4n =2 0 0 ;湘云鲫为 3n =15 0 ;红鲫和湘江野鲤分别为 2n =10 0。减数分裂时染色体分布情况与肾细胞染色体检测结果相吻合。具有四套染色体的异源四倍体鲫鲤在减数分裂中只形成 10 0个二价体 ,而不形成 2 5个四价体或其它形式 ,为产生稳定一致的二倍体配子提供了重要的遗传保障 ,也为人工培育的异源四倍体鲫鲤群体能够世世代代自身繁衍下去提供了重要的遗传学证据。三倍体湘云鲫在减数分裂过程中出现二价体、单价体共存 ,同源染色体在配对和分离中出现紊乱 ,导致非整倍体生殖细胞的产生 ,为湘云鲫的不育性提供了染色体水平上的证据     

小麦等离子束处理的生物学效应研究

本文以早熟品种冀麦31号和晚熟品系88—4284萌动种子为材料,报道了等离子束处理对小麦种子萌发的影响和细胞学效应。观察表明,等离子束长时间处理抑制种子发芽。在根尖细胞有丝分裂中期观察到染色体断裂和断片,冀麦31号染色体断裂的频率为0.49—1.34％,88—4284为0.21—2.14％。随着处理时间延长,染色体断裂频率逐渐提高。有丝分裂后期和末期出现大量的落后染色单体和染色体桥。在花粉母细胞减数分裂中期Ⅰ,经过等离子束处理的材料出现环状单价体。后期Ⅰ观察到染色体倒位造成的染色体桥和断片。在四分体期还有微核出现。     

黄瓜同源三倍体创制及减数分裂行为观察

采用常规杂交法研究黄瓜二、四倍体杂交过程中亲本育性、授粉组合及亲本基因型对杂交结实率的影响,并利用减数分裂制片法对获得的黄瓜同源三倍体进行了花粉母细胞减数分裂行为的观察.结果显示:(1)同源四倍体白交结实率比较低(13.0%～14.5%),可能与其花药内所包含的正常花粉粒比例小及花粉管萌发长度较短有关.(2)二、四倍体杂交组合的结实率很低(0.26%～0.02%),但在两种配组方式之间存在着明显差异,即以同源四倍体黄瓜为父本和二倍体黄瓜为母本的杂交结实率比较高,反之则杂交结实率比较低.(3)在二、四倍体杂交过程中,二、四倍体的基因型对杂交结实率的影响较大,以杂交双亲同属一个基因型的杂交效果较好.(4)同源三倍体花粉母细胞减数分裂过程与二倍体基本相同,但存在较高频率的染色体异常行为:中期I染色体构型复杂,在大多数花粉母细胞中可观察到单价体、二价体、三价体的存在;中期Ⅰ和Ⅱ有少数染色体游离于赤道板外;后期I和后期Ⅱ常出现落后染色体、染色体桥及细胞分裂不同步现象,其最终结果导致了不正常四分体和不可育配子的形成.(5)同源三倍体花粉粒的平均可染率和萌发率分别为18.8%和11.3%.研究结果表明,黄瓜二、四倍体正反交能直接获得同源三倍体材料;同源三倍体花粉母细胞减数分裂异常导致不育配子的形成是引起其育性低的细胞学原因;同源三倍体的部分育性为通过同源三倍体的回交来选育初级三体系奠定了基础.     

Cytogenetic studies on two F1 hybrids of autotetraploid rice varieties showing extremely high level of heterosis

Mechanisms of two F1 hybrids (D46A × DTP-4 and D46A × Dminghui63) of autotetraploid rice (2n = 4x = 48) showing extremely high pollen fertility 87.40% and 85.97%, respectively, seed set 82.00% and 79.00%, respectively and extremely high level of heterosis were analyzed cytologically. The chromosome pairing of D46A × DTP-4 and D46A × Dminghui63 was normal at metaphase I(MI), and had almost no I or III, with an average of 0.020I +14.36 II 6.44rod+7.91ring) +0.01III + 4.80 IV + 0.01VIII and 0.06 I + 17.67 II (11.01rod + 6.67ring)] + 0.06 III +3.10IV+0.01VI, respectively. The most frequent chromosome configurations were 10II+7IV and 12II+bIV. The bivalent frequency was less frequent in hybrids than that in restoring parents, and the same results were gained from univalents, trivalent and multivalents. However, the quadrivalent frequency was significantly higher in hybrids than that in restoring parents at MI. The other meiotic phases progressed normally, except for low percentages of PMCs with lagging chromosomes at AI and low percentages of PMCs with micronuclei at telophaseI (TI) and telophaseII (TII). PMCs with lagging chromosomes at AI and PMCs with micronuclei at TI and TII showed negative correlation between pollen fertility and seed set. Above 90% of the PMCs could form normal microspores, which resulted in the production of viable pollen grains, abnormal microspores were observed including penta-fission and hexa-fission. Based on these results we suggest that the two F1 hybrids had better behaviors of chromosome pairing and genetic stability than autotetraploid rice and other autotetraploid plants ever studied.     

Robertsonian translocations in wheat arise by centric misdivision of univalents at anaphase I and rejoining of broken centromeres during interkinesis of meiosis II

The mechanism of origin of Robertsonian translocations was investigated in plants monosomic for chromosome 1A of wheat and 1H(t) of Elymus trachycaulus by GISH. Chromosomes 1A and 1H(t) stayed univalent in all metaphase I cells analyzed, suggesting that Robertsonian translocations do not originate from meiotic recombination in centromeric regions with shared DNA sequence homology. At ana-/telophase I, the 1H(t) and 1A univalents underwent either chromosome or chromatid segregation and misdivided in 6-7% of the pollen mother cells. None of the ana-/telophases I analyzed had Robertsonian translocations, which were only observed in 2% of the "half tetrads" at ana-/telophase II. The frequency of Robertsonian translocations observed at ana-/telophase II corresponds well with the number of Robertsonian translocations (1-4%) detected in progenies derived from plants monosomic for group-1 chromosomes of wheat (1A, 1B, and 1D) and 1H(t) of E. trachycaulus. Our data suggest that Robertsonian translocations arise from centric misdivision of univalents at ana-/telophase I, followed by segregation of the derived telocentric chromosomes to the same nucleus, and fusion of the broken ends during the ensuing interkinesis.     

Asymmetric division of spindle microtubules and microfilaments during bovine meiosis from metaphase I to metaphase III

The kinetics of spindle and chromosomes during bovine oocyte meiosis from meiosis I to meiosis III is described. The results of this study showed that (1) oocytes began to extrude the first polar body (Pb1) at the early anaphase I stage and the Pb1 totally separated from the mother cell only when oocytes reach the MII stage; (2) the morphology of the spindle changed from barrel-shaped at the metaphase stage to cylinder-shaped at early anaphase, and then to a thin, long triangle-shaped cone at late anaphase and telophase stages; (3) chromosome morphology went from an individual visible stage at metaphase to a less defined chromatin state during anaphase and telophase stages, and then back to visible individual chromosomes at the next metaphase; (4) chromatin that connected with the floor of the cone became the polar bodies and expelled, and almost all of the microtubules (MTs) and microfilaments (MFs) composing the spindles moved towards and contributed to the polar bodies; and (5) the size of the metaphase I (MI) spindle was larger than the metaphase II (MII) and metaphase III (MIII) spindles. The MII spindle, however, is more barrel-shaped than the MI spindle. This study suggests that spindle MTs and MFs during bovine oocyte meiosis are asymmetrically divided into the polar bodies.     

A comparative cytogenetic study of the rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) autotetraploid restorers and hybrids

We studied pollen fertility, seed set and cytogenetic characteristics of restorer lines and F1 hybrids of autotetraploid rice. T4002, T4063, T461A × T4002 and T461A × T4063 showed significantly higher pollen fertility and seed set than T4132 and T461A × T4132. Meiotic pairing configurations of T4002, T4063, T4132, T461A × T4002, T461A × T4063 and T461A × T4132 were 0.05 I + 19.96 II (9.89 rod + 10.07 ring) + 0.01 III + 2.00 IV, 0.11 I + 19.17 II (8.90 rod + 10.37 ring) + 0.09 III + 2.26 IV + 0.01 VI, 1.34 I + 9.46 II (4.50 rod + 4.96 ring) + 0.80 III + 6.02 IV + 0.09 VI + 0.09 VIII, 0.02 I + 14.36 II (6.44 rod + 7.91 ring) + 0.01 III + 4.80 IV + 0.01 VIII, 0.06 I + 17.67 II (11.01 rod + 6.67 ring) + 0.06 III + 3.10 IV + 0.01 VI and 1.11 I + 11.31 II (5.80 rod + 5.51 ring) + 0.41 III + 5.63 IV + 0.03 VI + 0.03 VIII, respectively. Configuration 16 II + 4 IV and 12 II + 6 IV occurred in the highest frequency among the autotetraploid restorers and hybrids. Meiotic chromosome behaviors were less abnormal in the tetraploids with high seed set than those with low seed set. The hybrids had fewer frequencies of bivalents, univalents, trivalents and multivalents than the restorers, but higher frequency of quatrivalents than the restorers at MI. The frequency of univalents at MI had the most impact on pollen fertility and seed set, i.e., pollen fertility decreased with the increase of univalents. The secondary impact factors were trivalents and multivalents, and bivalents and quatrivalents had no effect on pollen fertility and seed set. The correlative relationship between pollen fertility and cytogenetic behaviors could be utilized to improve seed set in autotetraploidy breeding.     

WHAMM is required for meiotic spindle migration and asymmetric cytokinesis in mouse oocytes

WASP homolog associated with actin, membranes and microtubules (WHAMM) is a newly discovered nucleation-promoting factor that links actin and microtubule cytoskeleton and regulates transport from the endoplasmic reticulum to the Golgi apparatus. However, knowledge of WHAMM is limited to interphase somatic cells. In this study, we examined its localization and function in mouse oocytes during meiosis. Immunostaining showed that in the germinal vesicle (GV) stage, there was no WHAMM signal; after meiosis resumption, WHAMM was associated with the spindle at prometaphase I (Pro MI), metaphase I (MI), telophase I (TI) and metaphase II (MII) stages. Nocodazole and taxol treatments showed that WHAMM was localized around the MI spindle. Depletion of WHAMM by microinjection of specific short interfering (si)RNA into the oocyte cytoplasm resulted in failure of spindle migration, disruption of asymmetric cytokinesis and a decrease in the first polar body extrusion rate during meiotic maturation. Moreover, actin cap formation was also disrupted after WHAMM depletion, confirming the failure of spindle migration. Taken together, our data suggest that WHAMM is required for peripheral spindle migration and asymmetric cytokinesis during mouse oocyte maturation.     

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.     

Meiosis in allopolyploid Crepis capillaris. II. Autotetraploids.

Meiotic chromosome pairing of autotetraploid Crepis capillaris was analysed by electron microscopy of surface-spread prophase I nuclei and compared with light microscopic observations of metaphase I chromosome configurations. Prophase I quadrivalent frequencies are high in all three tetrasomes. (A, D, and C) and partially dependent on chromosome size. At metaphase I quadrivalent frequencies are much lower and strongly dependent on chromosome size. There is no evidence for multivalent elimination during prophase I in this system, and the reduction in multivalent frequency at metaphase I can be explained by an insufficiency of appropriately placed chiasmata. The high frequencies of prophase I quadrivalents far exceed the two-thirds expected on a simple model with two terminal independent pairing initiation sites per tetrasome, suggesting that multiple pairing initiation occurs. Direct observations reveal relatively high frequencies of pairing partner switches (PPSs) at prophase I, which confirms this suggestion. The numbers of PPSs per tetrasome show a good fit to the Poisson distribution, and their positional distribution along chromosomes is random and nonlocalized. These observations favour a model of pairing initiation based on a large number of evenly distributed autonomous pairing sites each with a uniform and low probability of generating a PPS.     

The fall of biological maturation promoting factor (MPF) and histone H1 kinase activity during anaphase and telophase in mouse oocytes.

Cell fusions have been used to determine the biological activity of the MPF complex in murine oocytes during their progression through anaphase and telophase to metaphase II. Oocytes (1) at metaphase I, (2) during the anaphase-telophase transition, or (3) at metaphase II were fused to germinal vesicle-staged (immature) oocytes. The hybrids were cultured for 1 h in the presence of db cAMP before fixation and nuclear evaluation. Metaphase I oocytes invariably induced germinal vesicle breakdown (GVBD) in the immature partner. By contrast, anaphase/telophase oocytes never induced GVBD in immature oocytes. The capacity to induce GVBD reappears after the formation of the second metaphase plate. In a second study, histone H1 kinase activity was measured during mouse oocyte maturation in single oocytes. H1 kinase activity was low in GV oocytes, increased sharply at MI, declined during anaphase and telophase and increased again at MII. After egg activation, H1 kinase activity was reduced to basal levels. These results provide direct evidence that a drop in activity of MPF in murine oocytes occurs concomitantly with the exit from metaphase I; MPF activity remains low until the cell re-enters metaphase.     

八倍体小滨麦与4D缺体杂交后代小染色体的传递与遗传效应

７２１８０ ４Ｄ缺体附加的１对小染色体（ｔｉ）为亚中部着丝点染色体,其大小为常染色体平均长度的１／３￣１／４。ＰＭＣ ＭＩ,染色体构型为１９．５９”（１８￣２０）＋０．４６’（０￣４）＋０．０９””（０￣１）＋０．９６ｔｉ”（０￣１）＋０．０８ｔｉ’（０￣２）,９６．１９％的细胞中,ｔｉ 色体联会成环状二价体,８２．３８％的ｔｉ”游离在赤道板两边,与常染色体不联会,且推后分离,与八倍体小滨麦杂交的     

利用中国春-山羊草2C二体附加系与中国春-偃麦草5E二体附加系杂交诱发染色体易位和缺失

通过对中国春－长穗偃麦草（E.elongata 2n=14EE)二体附加系与中国春－柱穗山羊草（Ae.cylindrica 2n=28CCDD)2C二体附加系（杀配子染色体）杂交F1减数分裂的观察,看到F1单价体数超过理论数值,后期出现大量的染色体段片,认为这种异常现象与杀配子染色体的作用有关。对76株中国春－长穗偃麦草5E二体附加系与中国春－柱穗山羊草2C二体附加系杂交、回交一代进行染色体C－分带鉴定,初步认定株系21－5－14、21－5－37、21－5－67、21－5－71为中国春染色体与偃麦草5E染色体易位。经染色体组原位杂交（GISH）进一步确定21－5－14为T5ES.4AS易位系,21－5－37为T5E.2BS易位系,21－5－67为T5E.3AS易位系,21－5－71为T5ES.5BS易位系。易位频率为5.3%。株系21－5－27、21－5－18、21－5－18、21－5－72、21－5－4、21－5－71经染色体C－分带鉴定,分别为6B、4B、5B、4A、4B、2A缺失系。杀配子染色体引起缺失频率为6.5%。证明利用杀配子染色体诱导染色体易位和缺失的频率较高。杀配子染色体引起中国春B组染色体畸变大于A、D组,与Endo观察结果相似。