玉米C型胞质雄性不育系花粉败育的细胞学研究

利用光学和电子显微镜对玉米C型胞质雄性不育系和保持系的花药与小孢子发育过程进行对比观察.结果表明,不育系小孢子初始败育发生在幼龄小孢子时期,单核中期基本上完全败育,绒毡层到开花前崩溃.在小孢子初始发育时期,不育系绒毡层细胞的质体呈“杯状”变态.导致雄性不育主要的细胞学原因是绒毡层细胞结构、功能的失常.     

大白菜CMS96不育系和保持系电镜观察

为了研究大白菜CMS96不育系花药败育机理,采用透射和扫描电镜,观察大白菜CMS96不育系和保持系的花药发育过程、花药和花粉表面结构。透射电镜结果表明,与保持系相比,在减数分裂期,大白菜CMS96不育系绒毡层与花药壁中层分开,不育系花粉母细胞皱缩,形状不规则,空隙较大,严重影响减数分裂正常进行;在单核小孢子时期,小孢子开始液泡化,直至成熟花粉期完全液泡化为空细胞,细胞壁严重萎缩,堆积在一起,最终花粉失去功能,导致败育。扫描电镜结果表明,大白菜CMS96不育系药室内的花粉畸形,花粉粒空瘪,表现为凹陷、扁平等形状,表面纹饰也呈多种不规则形状。花粉囊不开裂,没有花粉散出。初步认为,大白菜CMS96不育系花粉的败育从减数分裂开始到成熟花粉,伴随花粉发育整个过程。大白菜CMS96不育系药室内花粉粒虽具有花粉壁的结构,但扁平、空瘪,重叠堆积在药室壁上,失去了活力,无法散开。     

萝卜雄性不育小孢子发生的形态学研究

对萝卜雄性不育花进行形态学观察，发现两种类型的不育花，即花粉败育型和雄蕊萎缩型。比较研究了可育花及花种类型的不育花小孢子发育的细胞形态学特征，发现萝卜小孢子败育是个连续过程，在造孢细胞时期就已出现败育迹象，在小孢子发育过程中，绒毡层细胞出现液泡化、肥大、增生等异常现象。小孢子的败育与绒毡层细胞异常同步发生，小孢子一直信滞在单核期不再进一步发育，最终消失。另外，在正常可育小孢子发育过程中还偶尔观察到了不育花粉粒的存在。     

花椰菜雄性不育系小孢子发育过程及其POD活性

对花椰菜小孢子发育过程的细胞学观察和POD酶活性测定表明，雄性不育系花粉母细胞的减数分裂行为正常，小孢子败育发生在四分体形成以后，导致小孢子败育的直接原因是绒毡层细胞的肥大生长和高度液泡化，致使绒毡层与小孢子正常发育的协调关系遭到了破坏，不育花药的过氧化物酶活性高于其保持系。     

小麦D型细胞质雄性不育系和保持系小孢子发育的超微结构观察研究

对小麦D型细胞质雄性不育系和保持系小孢子发育的超微结构观察发现,D型不育系小孢子在“小液泡期”即表现出败育迹象．花粉败育过程中,小孢子液泡膜和细胞质膜断裂破碎,细胞质解体,线粒体、质体、内质网等细胞器解体或退化,绒毡层持续不解体,并缺少乌氏体的分泌。小孢子细胞解体顺序为：细胞膜首先断裂,细胞质分解变稀     

萝卜胞质紫菜薹雄性不育系花药发育的细胞形态学研究

通过2种细胞质紫菜薹雄性不育系小孢子发生的细胞形态学观察表明,供试不育系的雄蕊都深度退化,花药内无花粉。改良不育系小孢子在四分体前发育正常,在单核小孢子期,绒毡层异常膨大挤压小孢子,造成小孢子发育营养不良,引起小孢子败育。Ogura不育系的小孢子发育有所不同,它在孢原细胞分化期之前就已有53%败育,不形成花粉囊;形成花粉囊的孢原细胞大多只形成2个体积很小的的花粉囊,其中的小孢子发育与改良不育系的小孢子发育相似:单核早期绒毡层膨大挤压小孢子,使小孢子败育。     

春萝卜雄性不育系4-05A小孢子败育的细胞学观察

为了确定其花粉败育的时期和细胞学特点,采用石蜡切片法,对春萝卜雄性不育系4-05A及其保持系4-05B小孢子发生和花粉发育过程进行观察、比较。结果表明:不育系和保持系小孢子母细胞都能正常进行减数分裂,四分体可以释放出小孢子,不育系4-05A败育发生在单核靠边期,解体后的绒毡层原生质团急剧而彻底地进入花粉囊,渗入花粉母细胞周围,并与花粉母细胞黏连在一起。     

不结球白菜Ogura雄性不育花器官形态及败育细胞学的研究

对不结球白菜细胞质雄性不育系S2108A和Y3611A及其相应的保持系S2108B和Y3611B进行了花器官形态比较和花药发育细胞学观察,结果表明:两套不育系和保持系之间在花器官的多个性状上差异极显著,而两个不育系S2108A和Y3611A之间除花蕾大小、雌蕊长和花瓣长有差异外,其他均不显著,说明从不同不育源转育来的两个不育系在花器官外形上差异不大;两个不育系花药败育表现均起始于四分体时期,且由于绒毡层的液泡化和径向膨大,挤压四分体小孢子导致败育,表明败育与绒毡层的不正常发育有关.此外,与Y3611A不同,在不育系S2108A中还存在另一种败育形式,绒毡层细胞壁发生融合,成为紧贴药室壁的类似于变形型绒毡层的周原质团,部分细胞质流人药室腔,包裹粘连四分体小孢子,成为染色很深的不连续团块状物,花药败育;两不育系花药发育存在花粉囊数目的变异,且花粉囊发育时期不同步,个别花粉囊较正常花粉囊发育推迟,在不育系Y3611A中还有巨型花粉囊的现象.     

玉米C型胞质雄性不育系花药发育的细胞学观察

对3个 C 型不育系及其同型保持系和4个恢复系与这3个不育系组配的12个 F_1的花药发育过程进行了观察,以了解 C 型不育系雄性败育的细胞学机理。观察结果表明,不同核背景的不育系,其雄性败育的细胞学特点各异。Cms-C Mo17的花药绒毡层细胞为双核型。该层细胞在减数分裂的四分体时期开始液泡化,并呈辐射状膨大,至小孢子的单核前     

小麦D型细胞质雄性不育系和保持系小孢子发育的超微结构观察

对小麦D型细胞质雄性不育系和保持系小孢子发育的超微结构观察发现,D型不育系小孢子在“小液泡期”即表现出败育迹象。花粉败育过程中,小孢子液泡膜和细胞质膜断裂破碎,细胞质解体,线粒体、质体、内质网等细胞器解体或退化,绒毡层持续不解体,并缺少乌氏体的分泌。小孢子细胞解体顺序为：细胞膜首先断裂,细胞质分解变稀薄,然后是核膜断裂,细胞核降解,细胞质及细胞核降解物质充满整个药室,最后核仁解体。     

番茄花药离体培养中低温预处理对小孢子发育的影响

为明确低温预处理对番茄花药离体培养过程中小孢子发育的影响，进行了番茄花药离体培养的低温预处理试验，试验结果表明：离体培养条件下，雄核发育存在三条途径，即B途径、A-V途径和A-G途径，以B途径为主；随着培养时间的增加，小孢子逐渐退化；对番茄花药离体培养进行低温预处理，可在一定程度上延缓番茄小孢子退化，提早雄核发育，并增加参与雄核发育小孢子的比率，但参与B-途径的小孢子比参与A-途径的增加得多。     

光皮树小孢子发育及其对花药愈伤组织诱导的影响

为了构建光皮树花药组织培养技术体系,以光皮树优良无性系湘林G1小孢子不同发育时期的花药为外植体进行愈伤组织诱导,结合对其小孢子不同发育时期中花的形态特征观察和叶片中GA、IAA、ZR、ABA内源激素变化研究,综合分析小孢子不同发育期对花药愈伤组织诱导的影响。结果表明：光皮树优良无性系湘林G1小孢子在花粉母细胞、四分体、单核细胞和双核细胞4个时期的花药愈伤组织诱导具有显著差异,其中处于单核靠边时期能获得较高诱导率;花药愈伤组织诱导率与小孢子不同发育时期叶片中内源激素GA、IAA、ZR、ABA的含量存在一定的相关性,其中与ABA的含量成显著负相关性。     

芹菜雄性不育花药发育的细胞学观察

对芹菜花药发育过程的细胞学观察表明,雄性不育系花粉母细胞的减数分裂行为正常,可以形成单核小孢子并从四分体中释放出来,这以后由于绒毡层细胞提前解体,使小孢子得不到充足营养而发生败育。     

The origin of unreduced microspores due to aberrant cytokinesis in the meiocytes of potato and its genetic significance

Summary In 7 aneuhaploids of Solanum tuberosum and one clone of S. phureja the mechanism of unreduced microspore production has been studied. It has been observed that the unreduced microspores are produced not only by nuclear restitution at meiosis but also by aberrant cytokinesis in the meiocyte. The size and the number of nucleoli that are present in young microspores at the sporad stage give a fairly reliable indication of the ploidy level and the type of nuclear restitution that has occurred. These two aspects are helpful to deduce the meiotic events that have probably led to the formation of unreduced microspores in some cases. It has been found that triads and dyads are formed regardless of the type of nuclear division or restitution, indicating that cytokinesis is independent of karyokinesis to some extent. Cytological basis of unreduced microspore formation in diploid forms of potato appears to be more complex than it was supposed earlier.     

不同预处理对萝卜花药组织结构的影响

研究了花枝低温预处理及常、低温下离体花药的甘露醇和秋水仙素预处理对萝卜花药组织结构的影响。结果表明:4℃低温条件下处理枝条,处理1,3,5 d的花药绒毡层细胞出现从排列整齐到逐步退化的现象,药室中的小孢子染色也逐渐加深,7 d后花药壁细胞退化严重,小孢子数目则减少,故枝条低温预处理合适的时间为3~5 d。4℃条件下离体花药经甘露醇和秋水仙素预处理3 d后,药室中小孢子数目多,染色深,处理超过3 d,小孢子数目急剧减少,活力下降。25℃条件下甘露醇和秋水仙素预处理不利于花药结构的维持与小孢子活力的保持。     

Structural observations during androgenic microspore culture of the 4c1 genotype of Zea mays L.

Summary The capacity of the maize genotype 4c1 to regenerate microcalli and embryos from cultured microspores has been examined by comparing various cold pretreatments and culture media, using microspores and pollen at different stages of development. Viability of cultured cells was tested with FDA and their development was traced with light and fluorescence microscopy using DAPI as a nuclear dye.It was found that a pre-incubation of dissected flowers floating in a liquid nutrient medium at 8°C during 10–14 days was most successful for the induction of cell division. Among the developmental stages tested only the microspores appeared to regenerate. Subculture at 25°C in the same liquid medium, supplemented with 0.1 mg/l TIBA, gave highest rates of microspore division, i.e. up to 70% at 4 to 6 days of culture.All pathways described earlier for maize androgenic embryogenesis were observed within the 4c1 genotype. Symmetric divisions occurred in cultured microspores but most frequently asymmetric divisions lead to the formation of microcalli within 12 days of culture. In at least 60% of all dividing microspores cells were derived from the generative nucleus. Microcalli further developed either into loose or compact calli. Compact calli formed embryo-like structures.Abbreviations DAPI 4,6-diamidino-2-phenylindole - Dicamba 3,6-dichloro-2-methoxy benzoic acid - 2,4D 2,4 dichlorophenoxyacetic acid - FDA fluorescein diacetate - PAA phenylacetic acid - TIBA 2,3,5-triiodobenzoic acid - YP medium Yu-Pei basal salt medium     

中国白菜小孢子离体培养和双单倍体植株再生

摘 要: 采用略加修改的NLN培养基（无机大量元素1/2）和甘蓝型油菜小孢子的胚状体诱导方法,培养了5个中国菜用小白菜品种和1个中国大白菜品种的小孢子,进行了双单倍体的诱导。结果表明,不同品种的小孢子产胚量差异很大,在接种的6个基因型中,有5个诱导出胚,诱导成功率83.33％ 。其中的中萁青抗热605产胚量最高,是8.676个胚／10蕾。这些胚在添加了NAA和PP333（多效唑）的B5培养基发育成健壮的小植株。关于6个基因型的小孢子胚胎发生能力,还有待进一步研究。     

Prospects of polyploidizing cassava, Manihot esculenta Crantz, by unreduced microspores

Eleven interspecific hybrids of cassava with the wild relatives Manihot glaziovii, M. pseudoglaziovii. M. dichotoma and M. anomala were examined to detect unreduced microspores. The frequency of dyads and triads was counted, as well as pollen viability. Of these hybrids, the second generation progeny of cassava×M. glaziovii showed a high frequency of unreduced microspores (3.7%) permitting their selection for further manipulation of this character. An association of vegetatively-reproduced genotypes and the occurrence of unreduced microspores has also been observed.     

Influence of the stage for anther excision and heterostyly in embryogenesis induction from eggplant anther cultures

In this work we address two aspects of eggplant flower biology potentially involved on the efficiency of anther culture: the selection of the best floral stage to extract anthers for culture, and the effect of heterostyly in the identification of suitable buds and anthers. For 12 different accessions, we determined morphological criteria (length ranges) to identify buds and anthers enriched in vacuolate microspores and young bicellular pollen, the stages most responsive to embryogenesis induction. While these microspore/pollen stages were the most responsive when isolated and cultured in liquid medium, we observed that culture of anthers containing these stages is not the best choice. Instead, the highest response was found for younger anthers, containing mostly young and mid microspores. We analyzed eggplant anther walls and found that their particular thickness may be behind this apparent discrepancy, since they may delay the diffusion of inducing factors to the anther locule, reducing their effect over inducible microspores. Thus, the culture of younger anthers would allow for younger microspores to grow up to the inducible stages while factors are entering the locule. We also analyzed the embryogenic response of short and long-styled buds present in Cristal, a heterostylic cultivar. Our results demonstrated that each floral morph produced buds and anthers of different lengths, but equally useful for anther culture, since similar amounts of embryos were produced. The practical application of these results may improve the efficiency of anther culture not only in these cultivars, but in others also presenting thick anther walls and heterostyly.     

Fertile maize lines obtained from isolated microspores

Microspore response of three- way cross maize hybrid genotype 3AL/95 (Zea mays L.) was studied under simplified isolation and culture conditions. Fertile plant production was achieved through abundant plant regeneration. As a total, microspores of 160 tassels were inoculated and five sustainable microspore derived callus cultures (SMC) were obtained. Hybrid seeds (ML SC), which were produced by crossing of regenerates from two SMCs, gave rise to subsequent vigorous and fertile progeny. The response of the 3AL/95 and ML SC microspores was studied in three liquid culture media in order to improve the early viability of microspores. Them N6M medium provided better survival of cultured microspores (p = 5%) than the ppN6M/89 and the YPM-G media. The pH 5.8 in mN6M medium revealed significant increase (p = 1%) in microspore viability as compared to pH 3.0. The ML SC microspores showed higher viability(30%) on the first day of culture in the mN₆M than those of the3AL/95 (19%) but without improved rate of callus formation and plant regeneration. This revised version was published online in August 2006 with corrections to the Cover Date.