甘蓝型油菜隐性上位互作核不育材料1665花药败育的细胞学研究

采用石蜡切片方法,对甘蓝型油菜隐性上位互作核不育材料1665的可育株与不育株花药进行细胞学观察.结果显示:(1)不育株花药在花粉母细胞减数分裂时期出现异常,部分花粉母细胞细胞分裂相不均等分裂或分裂异常.导致部分四分体形状异常.(2)不育株绒毡层细胞在四分体时期开始生长膨大,单核花粉时期出现液泡化和巨型化,侵占药室,使得小孢子不能正常释放或无法继续发育;部分释放出的小孢子未及时形成花粉壁,阻碍花粉继续发育.不能发育形成二核期和三核期花粉,导致花药败育.     

长春蒲公英雄性不育株形态学观察与败育细胞学研究

在长春蒲公英(Taraxacum junpeianum Kitam.)株群中发现雄性不育现象,为研究其败育机理及特点,探寻其不育基因,采用形态观察法、石蜡切片技术和染色体压片法,对长春蒲公英野生型及其雄性不育株的花药发育过程和花粉母细胞减数分裂过程进行了观察。结果表明:(1)长春蒲公英雄性不育株花药中部发红、干瘪、无花粉散出。与野生型比较,雄性不育株雄蕊更短,子房更窄,种子形态更加狭长;(2)长春蒲公英雄性不育株败育时期为四分体到单核小孢子前期,败育方式为小孢子自身异常发育,绒毡层异常分解,互相粘连败育;(3)长春蒲公英雄性不育株花粉母细胞减数分裂二分体时期出现落后微核,随后产生极少四分体,并且四分体产生大量染色体桥,小孢子营养物质流失,彻底败育。因此,长春蒲公英雄性不育株败育彻底、稳定,并且有种的特点。小孢子自身异常发育和绒毡层异常分解是导致败育的主要原因。     

蓝猪耳小孢子发生和雄配子体发育

利用常规石蜡切片和超薄切片技术研究蓝猪耳(Torenia fournien)小孢子发生和雄配子体发育过程.蓝猪耳雄蕊4枚,花药具4个花粉囊.小孢子母细胞经减数分裂成四分体,其排列方式为四面体形或左右对称形.成熟花粉属2细胞型,具3个萌发孔.花药壁发育为双子叶型,腺质绒毡层.小孢子母细胞在四分体时期频繁出现细胞质降解的异常现象,其它发育阶段均正常;小孢子母细胞不正常的减数分裂可能导致花粉败育,这可能是蓝猪耳结实率低的原因之一.     

桔梗花粉母细胞减数分裂及雄性败育的细胞生理学研究

以桔梗不育系PA及其保持系PB为试验材料,采用石蜡切片和改良苯酚品红染色压片法对花粉母细胞减数分裂和雄配子体发育过程进行比较,探讨桔梗雄性不育系小孢子形成过程和败育发生的细胞生理学机理。结果表明:(1)桔梗保持系PB花粉母细胞减数分裂的细胞质分裂为同时型,同一花药减数分裂较同步;在中期Ⅰ和中期Ⅱ,少数细胞中可见赤道板外染色体;四分体以四面体为主,成熟花粉粒为二核花粉。(2)不育系PA花粉母细胞减数分裂后期Ⅰ开始出现异常,表现为细胞质形态改变,末期Ⅱ之后细胞质不能分裂,形成异常四分体,胼胝质壁不能溶解,四分体难以释放出游离小孢子而被降解,导致败育。(3)在发育过程中,桔梗不育系花蕾游离脯氨酸、可溶性蛋白含量低于保持系,而SOD活性、丙二醛含量均高于保持系。(4)桔梗不育系PA及其保持系PB花粉母细胞减数分裂和雄配子体发育过程存在明显差异,桔梗雄性败育过程大体可分为4个阶段,即后期Ⅰ细胞质异常、末期Ⅱ之后细胞质不能分裂、四分体难以释放游离小孢子、四分体被降解仅残留碎片。研究认为,桔梗不育花蕾(开花前)生长发育过程中,体内活性氧代谢紊乱、丙二醛积累及游离脯氨酸等"物质代谢损亏"可能是引起桔梗雄性败育的原因。     

湿地松雄性不育和可育系小孢子叶球形态和细胞发育动态变化

为了解湿地松‘松泰’小孢子叶球在发育过程形态是否有差异变化,明确其败育过程、败育方式及影响因素,为湿地松雄性不育品种利用和后期开展相关研究提供科学依据。该研究以‘松泰’s10败育系和s9可育系为材料,观察小孢子叶球形态发育变化,并对其小孢子叶球进行石蜡切片,在光学显微镜下观察小孢子发育过程。结果表明:s10败育系和s9可育系在小孢子母细胞减速分裂前无明显差异,小孢子叶球生长趋势也一致;四分体时期,s10小孢子细胞发育异常,小孢子叶球形态发育也出现异常,二者异常发育具有同步性;可育系从四分体到单核小孢子发育阶段的时间为5 d左右,而败育系持续发育长达20 d左右,持续时间为可育系的4倍。在此期间出现小孢子绒毡层细胞发育异常、降解缓慢,小孢子囊壁组织排列紊乱、降解延迟等现象,s10形成异常二核花粉,且无花粉散出。因此,推论s10小孢子败育的原因主要是小孢子囊壁细胞发育异常,其小孢子叶球形态异常,相对应的绒毡层在四分体时期发育异常,不能适时地分泌胼胝质酶来降解围绕着四分体的胼胝质壁,也不能适时地合成输送花粉形成所需能量物质,同时囊壁细胞出现降解延迟和层积,这一系列的异常变化导致不能形成正常四分体,从而使花粉败育。     

玉米新选细胞质雄性不育系小孢子发育的细胞学观察及DNA甲基化分析

细胞质雄性不育在高等植物中普遍存在,是杂种优势利用的重要工具,为推动植物杂种优势的利用发挥了重要作用。文章以本课题组前期新选的玉米细胞质雄性不育系A1、A2及保持系18(红)为材料,利用石蜡切片技术对不育材料小孢子发育过程进行细胞学观察,采用高效液相色谱法(HPLC)对不同发育时期的叶片及不同发育时期的雄穗DNA进行甲基化分析,从细胞学和表观遗传学角度了解不育系A1、A2的败育机制。结果表明：不育材料A1、A2小孢子发生败育的主要时期为四分体时期至单核小孢子中期。在不育系A2中还存在另一种败育方式,即在花粉母细胞时期表现出败育特征。甲基化分析结果表明,保持系18(红)的叶片DNA甲基化水平从苗期到拔节期迅速上升,而不育系A1、A2叶片DNA甲基化水平基本保持不变;保持系雄穗DNA甲基化水平表现为从花粉母细胞时期到双核期逐渐升高,而不育材料A1、A2从花粉母细胞时期到双核期的雄穗DNA甲基化水平表现为先上升后下降的趋势,达到最高峰的时期均出现在小孢子发育的四分体时期。从小孢子发育的细胞学观察结果可以发现,小孢子败育的主要时期往往具有较高的甲基化水平,推测DNA甲基化水平变化可能与不育材料A1、A2的花粉败育有关。     

红菜薹雄性不育系花药败育的细胞形态学观察

采用石蜡切片技术,在光学显微镜下系统研究了红菜薹(Brassica campestris L.ssp．chinensis L.var．utilis TsenetLee．)波里马胞质雄性不育系(Polima CMS)、红菜薹萝卜胞质雄性不育系(Ogura CMS)及相应保持系花药发育过程的细胞形态学特征。观察结果表明：红菜薹Polima CMS花药发育受阻于孢原细胞阶段,不形成花粉,属无花粉型,此不育系花药不形成绒毡层和中层;而红菜薹Ogura CMS花药败育发生于小孢子母细胞期或四分体时期,表现为绒毡层细胞异常,挤压四分体,导致四分体和绒毡层同时解体而败育。     

OT杂种系百合品种‘Cocossa’小孢子败育细胞学研究

以OT百合‘Cocossa’为材料,采用醋酸洋红染色烤片法和石蜡切片法,对该品种花粉母细胞减数分裂过程以及绒毡层在花粉母细胞发育过程中的变化进行系统的细胞学观察。研究发现:(1)当百合花蕾长度在2.5~4.5 cm时,花粉母细胞处在减数分裂时期。(2)在花粉母细胞减数分裂的过程中,出现了较多染色体异常现象:中期Ⅰ出现染色体环;后期Ⅰ出现了滞后染色体和染色体桥;在末期Ⅰ和末期Ⅱ细胞出现不均等分裂和微核;处于同一花蕾和同一花药的花粉母细胞减数分裂具有不同步性,能同时观察到2个不同时期的分裂相;形成畸形、无核、多核的异常小孢子。(3)在末期Ⅱ和单核花粉粒时期,绒毡层延迟降解,引起多核花粉粒、畸形花粉粒以及花粉粒自溶现象。结果表明:OT百合‘Cocossa’小孢子败育是花粉母细胞异常减数分裂与绒毡层异常降解共同导致的。     

温光敏核雄性不育小麦雄性败育的细胞学观察

为了掌握小麦温光敏核雄性不育系雄性败育发生的时期和类型,为两系法利用小麦杂种优势提供依据,用涂抹压片法观察了其减数分裂和小孢子发育过程.结果表明,在低温短日照条件下,在叶枕距-5cm 左右、幼穗长5cm 左右、孕穗前5～10d的时段处于花粉母细胞形成期,发育正常;叶枕距-2～1cm 、幼穗长8cm 左右、孕穗前4～5d到孕穗当天的时段处于减数分裂期,减数分裂行为基本正常,能形成正常的四分体;叶枕距5cm 左右、穗长9cm左右、孕穗后到抽穗前的时段处于小孢子发育期,小孢子能发育到单核靠边期,但此后就发育停滞,细胞质和核物质逐渐解体,不能被苏木精或碘液染色,出现败育;发生败育的高峰在小孢子单核晚期,败育花粉的主要类型是圆败型.而在高温长日照条件下,减数分裂和小孢子发育过程与常规对照品系相同,能形成正常可育的三核花粉粒.     

灯盏花雄性不育种质鉴定与花药发育的细胞学研究

对云南泸西栽培灯盏花群体进行调查,发现了灯盏花雄性不育种质个体,其出现频率约为1.06×10-4.对所发现的灯盏花不育株形态特征及其花药发育过程进行了观察,并对花粉活力进行鉴定.结果显示:(1)灯盏花不育株根、茎、叶形态与正常可育植株基本相似,管状花小,花丝短,花药瘦小,无花粉粒散出或花粉无活力.(2)灯盏花在其花药发育的小孢子母细胞时期、四分体时期、小孢子时期和单核早期,由于绒毡层细胞液泡化、提前解体,不能为小孢子或花粉发育提供所需物质,导致小孢子母细胞和四分体解体,产生无花粉的花药;或小孢子和单核花粉胞内降解,形成不同形状和外壁纹饰的败育花粉.研究认为,灯盏花花药绒毡层异常是其花粉败育的主要原因.     

融安金柑雌雄配子发育相关性及2n配子诱导研究

以融安金柑为试验材料,观察了2n花粉自然发生率、雌雄配子发育过程及减数分裂行为,以不同秋水仙素浓度及处理时间对花蕾进行2n配子诱变.结果显示:(1)融安金柑2n花粉自然发生率为0.31％;(2)花蕾纵径为0.228～0.270cm时,为花粉母细胞减数分裂Ⅰ前期;花蕾纵径为0.274～0.318 cm时,大孢子处于减数分...     

欧洲黑杨花粉母细胞减数分裂观察及其不同步性分析

以欧洲黑杨的雄花枝为材料,采用温室切枝水培和醋酸洋红染色制片法,观察花粉母细胞减数分裂进程及其与花芽发育外部形态的关系。结果表明:(1)欧洲黑杨花粉母细胞的减数分裂进程快慢与温度高低关系密切;(2)减数分裂发生时其花芽的外部形态发育没有明显变化;(3)花粉母细胞减数分裂不同步性突出,同一花药内的花粉母细胞处于不同的减数分裂时期,且同一花芽尖端发育明显滞后于中部和基部;(4)减数分裂过程中有少量的核仁异常现象,虽观察到较多平行纺锤体现象,但未发现天然2n花粉。该研究结果揭示了欧洲黑杨减数分裂的遗传变异规律,为诱变育种、倍性育种以及分子水平的进一步研究奠定了细胞学基础。     

关苍术两性花与雌花花药的解剖学研究

苍术属(Atractylodes DC.)是菊科菜蓟族(Cynareae)刺苞亚族(Carlininae O. Hoffm.)的一个东亚特有属,世界上仅有7种,其中我国有5种。该研究以关苍术为材料,采用石蜡切片法比较研究了两性花和雌花的花药及雄配子体发育进程,并进一步探讨了其雌花产生花药退化的时期及原因。结果表明：(1)关苍术小孢子发育与花蕾长度间存在相关性,当花蕾长度在5 mm时进入花粉母细胞时期,花药壁已分化,在7~9 mm时处于四分体时期,大于11 mm时开始进入花粉粒时期。(2)关苍术花药5个,花粉囊4个,减数分裂属同时型,四分体以正四面体为主,属3-细胞型,萌发沟3个。(3)关苍术花粉囊壁发育属双子叶型,从外层的表皮、药室内壁,到内层的中层和绒毡层均由一层细胞构成,关苍术绒毡层为腺质绒毡层。(4)关苍术雌花花药退化发生在花药发育早期至四分体时期,表现为花药发育早期畸形、药壁分化异常、小孢子母细胞发育停滞在前期、绒毡层增生4个原因。该研究结果为苍术属植物的系统发育、物种形成和进化提供胚胎学依据。     

Microsporogenesis of the feathers (fertile branches derived from annual buds) in Vitis vinifera,cv Picolit giallo

Abstract

Using light and electron microscopy, we have studied the microsporogenesis and tapetal development of the feathers in two different low producing clones of Picolit giallo (sp. Vitis vinifera). In these clones while the productivity of the main branches (fertile branches originated from buds, formed in the previous year, that remained silent during the winter) is very low, that of the feathers (fertile branches derived from annual buds) is always normal.

The microsporogenesis and tapetal development proceed normally in almost all the examined anthers; it is remarkable that at the tetrad stage the tapetal cells appear well structured without any degeneration symptom, unlike what observed for the main branches. Moreover in most of the mature anthers the pollen grains are numerous, pleinty of organelles and show sometimes thickenings in the callose layer under their wall. The tapetal cells of these anthers have disappeared. Only in few anthers we observed the presence of collapsed pollen grains and tapetal cells with anomalous development, that are still present when the pollen grains are mature. This rare situation for the feathers is on the contrary frequent for the main branches.     

温度对斯托克通氏烟草雄配子体形成和发育的影响

为探究低温对斯托克通氏烟草(Nicotiana stocktonii)花粉母细胞(PMC)减数分裂及其雄配子体发育过程的影响,采用卡宝品红染色法,研究不同温度条件下该材料雄配子体形成和发育的过程。结果表明:种植于昼温(31±0.5)℃、夜温(11±0.5)℃人工气候箱中的Nicotiana stocktonii花粉母细胞减数分裂过程异常现象较少,出现微核的比率较低,用新鲜成熟的花粉做萌发实验花粉萌发率较高,为(71±3)%; 而种植于昼温(25±0.5)℃、夜温(3±0.5)℃条件下的Nicotiana stocktonii开花后花药大多干瘪,用新鲜成熟花粉做萌发实验花粉萌发率低,为(13.67±3)%,花粉母细胞减数分裂过程出现染色体桥、染色体不同步、染色体断片、落后染色体等现象,存在微核的细胞比率较高。因此,Nicotiana stocktonii花粉母细胞减数分裂与小孢子发育过程易受温度影响,从而影响花粉的可育性。     

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.     

Tetrad pollen formation in quartet mutants of Arabidopsis thaliana is associated with persistence of pectic polysaccharides of the pollen mother cell wall

The quartet (qrt) mutants of Arabidopsis thaliana produce tetrad pollen in which microspores fail to separate during pollen development. Because the amount of callose deposition between microspores is correlated with tetrad pollen formation in other species, and because pectin is implicated as playing a role in cell adhesion, these cell-wall components in wild-type and mutant anthers were visualized by immunofluorescence microscopy at different stages of microsporogenesis. In wild-type, callose was detected around the pollen mother cell at the onset of meiosis and around the microspores during the tetrad stage. Microspores were released into the anther locule at the stage where callose was no longer detected. Deposition and degradation of callose during tetrad pollen formation in qrt1 and qrt2 mutants were indistinguishable from those in wild-type. Enzymatic removal of callose from wild-type microspores at the tetrad stage did not release the microspores, suggesting that callose removal is not sufficient to disperse the microspores in wild-type. Pectic components were detected in the primary wall of the pollen mother cell. This wall surrounded the callosic wall around the pollen mother cell and the microspores during the tetrad stage. In wild-type, pectic components of this wall were no longer detectable at the time of microspore release. However, in qrt1 and qrt2 mutants, pectic components of this wall persisted after callose degradation. This result suggests that failure of pectin degradation in the pollen mother cell wall is associated with tetrad pollen formation in qrt mutants, and indicates that QRT1 and QRT2 may be required for cell type-specific pectin degradation to separate microspores.     

小叶丁香引种栽培条件下种子败育的解剖学研究

为探讨小叶丁香在北京栽培条件下种子败育的问题,对其胚胎发育过程进行了解剖学观察研究。结果显示:(1)小叶丁香人工异花授粉坐果率为20.37%,天然授粉坐果率为9.13%,人工自花授粉以及去雄套袋授粉均未坐果;各种授粉方式结实率均为0。(2)小叶丁香花药壁发育为基本型,腺质绒毡层,胞质分裂为同时型,小孢子四分体排列方式以四面体型为主,2-胞成熟花粉;倒生胚珠,单珠被,薄珠心,直线型大孢子四分体,胚囊发育为蓼型,核型胚乳,胚发育为紫菀型,先后形成球形胚、心形胚、鱼雷形胚和子叶胚;9月1日以后,子叶胚发育停滞,细胞逐渐降解。研究表明,栽培条件下,小叶丁香为异花授粉植物,人工异花授粉可提高植株坐果率,但未获得种子;其大小孢子及雌雄配子体发育、传粉受精过程、胚胎发育初期均正常,但后期发育停止,是小叶丁香引种栽培条件下种子败育的关键环节。     

Flower developmental stage and organ sensitivity of bell pepper (Capsicum annuum L.) to elevated temperature

High temperatures adversely affect crop productivity of several plant species including bell pepper (Capsicum annuum L. var. annuum). The objectives of this study were: (1) to determine whether flower ontogeny is adversely affected by high temperature during different phases of development, including pre‐ and post‐pollination events; (2) to determine the duration of high temperature exposure necessary to cause reduction in fruit set; and (3) to determine whether injury to the pistil or stamen during development is responsible for reduced fruit set during high temperature. We determined that flower buds at <2·5 mm in length, corresponding to microspore mother cell meiosis to tetrad dissolution, and flowers that reached anthesis during the high temperature exposure had reduced fruit set when exposed to 33 °C for 48 or 120 h. Flower buds at <2·5 mm in length also had reduced pollen viability when exposed to 33 °C for 120 h. Morphological examination demonstrated that meiocytes initiated tetrad formation, but after dissolution the microspores remained small and clumped without a thick exine. High temperature exposure at a late‐development, pre‐anthesis stage did not affect pistil or stamen viability, but high post‐pollination temperatures inhibited fruit set, suggesting that fertilization is sensitive to high temperature stress.     

两种山羊豆属植物花粉形态与花粉母细胞减数分裂行为观察

采用扫描电镜与1%醋酸洋红压片的方法,观察山羊豆属两种植物的花粉形态及花粉母细胞减数分裂行为,从而为山羊豆属的遗传学及分类学提供依据。两种山羊豆属植物立体形状为近球形或扁球形,均具三孔沟,穴—网状纹饰,极面观为近三角形,赤道面观为圆形。东方山羊豆的大小为:赤道轴×极轴=14.3（13.5～15.0）×12.6（12.1～13.7）μm,山羊豆的大小为：赤道轴×极轴=13.7（12.3～14.8）×12.4（10.6～13.0）μm.花粉母细胞减数分裂为同时型胞质分裂,终变期染色体构型为8Ⅱ,没有出现落后染色体及微核现象。小孢子四分体为四面体型。减数分裂过程有一定规律,与花蕾大小,颜色有密切关系。减数分裂进程存在一定差异,同一个花蕾中的不同花药花粉母细胞分裂速度基本相同,所处的时期及观察的分裂相一致,在同一个花药中的不同花粉母细胞中,可见各种不同时期的细胞。