青花菜小孢子发育时期与花器形态的相关性

对青花菜小孢子发育时期细胞学特征及其与花器外部形态的相关性展开研究,试图从花器的外部形态特征来推断小孢子的发育时期,最终目的是为花药或游离小孢子培养研究提供科学依据。结果表明,青花菜小孢子发育经历四分体时期、单核早中期、单核靠边期和双核期共4个时期,供试3个青花菜各时期细胞学特征存在明显差异。青花菜小孢子发育时期与花蕾大小和花药颜色等指标密切相关。供试青花菜花蕾纵径为10.52~11.05 mm,花蕾横径为3.64~4.25 mm,花药长度为2.85~2.87 mm,花药宽度为0.95~0.96 mm,且花瓣微露出花萼,花瓣和花药均为淡黄色时,80%以上的小孢子发育至单核靠边期。     

火龙果小孢子不同发育时期的形态特征

阳现蕾起观测"紫红龙"火龙果花蕾发育的动态变化,采集从现蕾期起1至13天的花蕾,通过电子显微镜观察火龙果小孢子在不同发育时期的外部形态结构特征及细胞学特征变化,为花药或小孢子培养提供细胞学依据和最佳取材时间,以期建立高效、稳定的火龙果花药或小孢子组织培养体系。结果表明,火龙果小孢子发育经历了四分体时期、单核早期、单核靠边期和双核期,最后发育为成熟花粉粒,每个时期具有明显不同的形态特征;火龙果小孢子发育时期与花蕾的大小和花药的长度紧密相关。火龙果花蕾纵径在5.5~6.0 cm,横径在2.5~3.2 cm,绝大数小孢子发育至四分体时期,此时对应的花蕾发育天数约为4 d;花蕾纵径在6.8~7.6 cm,横径在3.0~3.4 cm,绝大数小孢子发育至单核早期,此时对应的花蕾发育天数约为5 d;花蕾纵径在8.1~8.6 cm,横径在3.3~3.9 cm,绝大数小孢子发育至单核靠边期,此时对应的花蕾发育天数约为6 d;双核期时,花蕾纵径在8.9~10.5 cm,横径在3.8~4.4 cm,对应的花蕾发育天数约7~8 d。因此,可以根据火龙果花蕾的形态、大小和花药发育时期,从而确定小孢子最佳培养时期对应的选蕾标准。     

蒺藜苜蓿小孢子不同发育时期花蕾形态与花药脱分化能力的研究

本研究以豆科模式植物蒺藜苜蓿为材料,研究其小孢子不同发育时期与花蕾形态的相关性,以及花药脱分化能力。结果表明：蒺藜苜蓿小孢子发育可分为单核早中期、单核靠边期和双核期,不同时期的小孢子与花蕾外部形态具有明显相关性。根据花蕾形态判断小孢子不同发育时期并进行培养,其中单核靠边期小孢子的花药愈伤组织诱导率最高,可达37.21%;单核早中期为4.82%,而双核期小孢子的花药无脱分化能力。这说明小孢子单核靠边期是蒺藜苜蓿花药培养的最佳时期,此时期的花蕾膨大,萼片绿色且比花瓣稍短。     

甜椒小孢子发育的细胞学观察

为研究甜椒小孢子发育时期的染色鉴定方法以及探讨小孢子发育时期与花器形态的相关性,采用不同染色方法进行甜椒小孢子的细胞学观察,并对小孢子发育期的细胞学特征以及小孢子不同发育时期与花器外观形态的关系进行研究。结果表明,甜椒小孢子经酒精盐酸处理后,用醋酸洋红染色液的染色效果最佳;甜椒小孢子的发育经历四分体期、单核早期、单核中期、单核后期、双核期直至花粉粒成熟期,各时期的特征明显;甜椒小孢子各发育时期与花蕾外部形态具有密切的相关性。     

黑皮冬瓜小孢子发育与花蕾形态建成的相关性研究

对4个黑皮冬瓜品种的小孢子发育进行了细胞学观察,并比较分析了小孢子发育与花蕾形态建成的相关性。结果表明,4个黑皮冬瓜品种的小孢子发育过程相同,可分为四分体时期、单核居中期、单核靠边期、单核有丝分裂期和双核期5个时期。小孢子发育与雄花蕾形态建成密切关联,小孢子直径与蕾径、蕾长、萼长、瓣长及雄蕊直径均呈对数式的正相关。因此,黑皮冬瓜小孢子发育时期可以根据雄花蕾的形态指标判定。黑皮冬瓜小孢子发育处于单核靠边期的雄花蕾蕾长0.54~0.76 cm,蕾径0.53~0.73 cm,萼长0.78~1.18 cm,瓣长0.49~0.60 cm,雄蕊直径0.39~0.53 cm,蕾长/蕾径0.99~1.06,萼长/瓣长1.68~1.97,着生于开花节之上的第7~8节。     

白芷小孢子发育时期与花器形态相关性分析

目的：研究3个白芷品系小孢子发育时期与花蕾形态和花药形态的相关性,以期直接从花蕾形态和花药形态判断小孢子的发育时期。方法：采用苯酚品红对白芷小孢子进行染色,对白芷小孢子发育的细胞学以及小孢子不同发育时期与花柄长、花蕾大小、花药直径、花蕾和花药颜色的关系进行研究。结果：白芷小孢子发育经过四分体时期、单核前期、单核中晚期、二核期和三核期共5个时期,各时期特征明显;3个白芷品系小孢子发育时期与花蕾的外部形态和花药颜色密切相关,其中以CX08-2的花柄长、花蕾直径和花药长度最大,其次为B-2,而B-6为最小;但单核中晚期与其相邻时期的花柄长、花蕾直径和花药长度未达到显著水平,花瓣颜色、花瓣是否开裂和花药颜色可以作为花药培养取材的直观依据。结论：通过花器官形态判断小孢子的发育时期可以为白芷花药培养接种材料的选择提供依据。     

甘蓝型油菜不同播期与小孢子发育状态荧光染色的研究

研究不同播种时期与小孢子发育状态的关系。在同一时间对不同播期及同一播期的花蕾进行荧光染色镜检,结果表明：不同播种时期,同样大小花蕾的发育状态不尽一致,早播材料处于单核晚期至双核早期的花蕾小,晚播的材料处于单核晚期至双核早期的花蕾稍大。通过建立花蕾模型图可以看出,播期不同,处于单核晚期至双核早期的花蕾长为2.0-4.0mm。在进行小孢子游离培养时,应该根据小孢子实际的发育状态来进行取蕾培养。     

金莲花花蕾大小与花粉发育时期的关系

本研究以张家口地道性中药材金莲花花蕾为材料,利用醋酸洋红染色剂染色的方法,通过在显微镜下观察花粉的发育情况,探讨出金莲花花蕾大小与花粉发育时期的关系。结果表明:当花蕾长度在5.32~7.32 mm和7.33~9.33 mm时,花粉大部分处于四分体时期,个别处于单核早期;长度为9.34~11.34 mm时,花粉发育时期为单核早中期;长度为11.35~13.35 mm时,花粉发育时期集中在单核后期;长度为13.36~15.36 mm时,此时的花蕾即将开放,为双核期。通过观察金莲花外观形态,确定其发育时期,为后期单倍体培养提供理论依据。     

朝鲜白头翁小孢子发育时期与花器形态的相关性研究

为了寻找朝鲜白头翁小孢子发育时期的鉴定方法以及探讨小孢子发育时期与花器形态的相关性,采用常规石蜡切片法,对朝鲜白头翁小孢子发育时期的细胞学以及小孢子不同发育时期与花蕾大小的关系进行了研究。结果表明：朝鲜白头翁每个花药具4个花粉囊,小孢子母细胞减数分裂属同时型,小孢子在四分体中的排列属四面体型。成熟花粉粒属2-细胞型,并有3个萌发沟。花粉囊壁由4层细胞构成,即表皮（1层）、药室内壁（1层）、中层（1~2层）、绒毡层（1层）。绒毡层发育属腺质型（或分泌型）,出现多核现象。朝鲜白头翁小孢子发育时期与花蕾的外部形态特征、花药颜色有一定的相关性。小孢子处于单核期时花蕾长度在1.10~1.20 cm,花萼外侧颜色为紫红色,花药的颜色为黄色,花丝的长度为0.4~0.5 cm。朝鲜白头翁同一花蕾不同花药小孢子的发育存在一定程度的渐续性。     

番茄小孢子发育时期鉴定和游离方法的研究

为寻找番茄小孢子单核期的鉴定方法以及将小孢子从花药中游离出来而不影响其活性的最佳方法,采用醋酸洋红染色法、酒精-盐酸-醋酸洋红染色法、DAPI 染色法,分别染小孢子单核期细胞核,pH6.8-8.0的中性红染小孢子单核期的液泡;采用挤压法、挤压离心法、自然散出法,分别从花药中游离出小孢子。实验结果醋酸洋红染细胞核着色不好,酒精-盐酸-醋酸洋红染细胞核着色清晰,DAPI染色细胞核清晰,但成本高且受时间限制,中性红染色液泡效果好,但受细胞活性影响;挤压法游离小孢子提前脱壳率较高,挤压离心法游离小孢子提前脱壳率最高,自然散出法游离小孢子脱壳率最小。所以,采用酒精-烟酸-醋酸洋红法鉴定番茄小孢子发育时期效果最好,采用自然散出法游离番茄小孢子可减少小孢子脱壳率,提高小孢子活性。     

长白山区紫草小孢子发生的细胞形态学观察

采用常规石蜡切片法,观察了长白山区紫草的小孢子发生过程和各时期的形态特征.结果表明:长白山区紫草小孢子发育直到四分体为止发育都正常,小孢子母细胞减数分裂为同时型,四分体排列方式为四面体型.小孢子发育受阻于四分体分离后的单核小孢子时期开始,是因为这时期绒毡层细胞异常膨胀,占满整个药室,小孢子没有可发育的空间,最终导致小孢子败育.     

Functional genomics of microspore embryogenesis

Isolated plant microspores, when stressed and cultured in vitro, can be diverted from their normal gametophytic pathway towards sporophytic development, with the formation of haploid embryos and ultimately doubled-haploid plants. This process is called androgenesis or microspore embryogenesis, and is widely used in plant breeding programmes to generate homozygous lines for breeding purposes. Protocols for the induction of microspore embryogenesis and the subsequent regeneration of doubled haploid (DH) plants have been successfully developed for more than 200 species. These practical advances stand in stark contrast to our knowledge of the underlying molecular genetic mechanism controlling this process. The majority of information regarding the genetic and molecular control of the developmental switch from gametophytic to sporophytic development has been garnered from four intensely studied (crop) plants comprising two dicotyledonous species, rapeseed (Brassica napus) and tobacco (Nicotiana tabacum), and two monocotyledonous species, wheat (Triticum aestivum) and barley (Hordeum vulgare). In these species the efficiency of microspore embryogenesis is very high and reproducible, making them suitable models for molecular studies. In the past, molecular studies on microspore embryogenesis have focussed mainly on the identification of genes that are differentially expressed during this developmental transition and/or early in embryo development, and have identified a number of genes whose expression marks or predicts the developmental fate of stressed microspores. More recently, functional genomics approaches have been used to obtain a broad overview of the molecular processes that take place during the establishment of microspore embryogenesis. In this review we summarise accumulated molecular data obtained in rapeseed, tobacco, wheat and barley on embryogenic induction of microspores and define common aspects involved in the androgenic switch.     

辣椒花药培养研究进展

归纳了辣椒花药培养过程中影响雄性胚胎发生的诸多因素,包括供试材料、小孢子发育时期、基本培养基、植物生长调节剂、培养基添加物质和温度胁迫处理,总结了小孢子胚胎发生的细胞学观察、再生株的倍性水平和单倍体基因组加倍研究,指出尽管人类已从细胞和分子方面对辣椒花药培养中诱导小孢子胚胎发生和形成胚状体有了进一步的认识,但尚未完全了解小孢子是如何被激发进入孢子体发育途径。当小孢子胚胎发生之谜完全揭开后,辣椒花药培养技术将会应用于更加广泛的领域。     

Effect of donor plant age and inflorescence age on microspore culture of Brassica napus L.

Summary Effect of age of donor plants and age of inflorescence on embryogenesis in microspore culture of B. napus was examined. Microspores isolated from buds of older plants had a higher embryo yield than those of younger ones. The effect of the age of inflorescence showed a different pattern. In older plants, a higher embryogenesis response was observed in microspores isolated from buds of new inflorescences, while in young plants, microspores isolated from buds of old inflorescences showed high embryo yield. These different responses were considered to be attributable to a difference in the developmental stage of pollen at the time of microspore isolation. Our results indicated that microspores collected from older inflorescences and older plants have sufficient embryogenic potential when the optimum developmental stage of pollen was used. Frequency of embryo to plant conversion was influenced by the size of embryos subcultured, but not by donor plant age or the age of the inflorescence.     

Efficient production of callus-derived doubled haploids through isolated microspore culture in eggplant (<Emphasis Type="Italic">Solanum melongena</Emphasis> L.)

Production of doubled haploids (DHs) through androgenesis induction is an important biotechnological tool for plant breeding. In some species, DHs are efficiently obtained through embryogenesis from isolated microspore cultures. In eggplant, however, this process is still at its infancy, despite the economic relevance of this important agricultural crop. To date, only two studies have focused previously on this process, suggesting that in eggplant microspore cultures, the only morphogenic response is callus formation. Given the notable lack of studies on eggplant microspore cultures, in this work we explored this process with different experimental approaches. We studied the response of different cultivars and characterized the development of microspores induced to divide and proliferate. We demonstrated that microspore-derived embryos (MDEs) can be produced in eggplant; however, MDEs stopped at the globular stage, to turn into euploid and principally mixoploid calli. From these calli, 60 % of DH plants could be regenerated. In order to promote microspore induction we evaluated the effect of polyethylene glycol (PEG) and mannitol. PEG, but not mannitol, significantly increased induction of microspore embryogenesis. We also tested the ability of eight different media compositions to promote efficient plant regeneration from calli. In order to test it in a genotype-independent manner, we previously developed a method to generate clonal callus populations derived from single microspore-derived calli. Together, the results presented hereby constitute an efficient way to produce eggplant DHs through microspore culture. In addition, they contribute significant insights into the knowledge of the particularities of androgenesis induction in this species.     

影响芦笋产量性状的相关性分析

对芦笋的6个农艺学性状指标进行相关性分析,并对产量和一级笋率进行了多重比较。结果表明,影响芦笋产量的重要因子有茎粗、株高和茎数。影响芦笋一级笋率的重要因子有茎粗、株高、产量。植株越高,产量越高,一级笋率也越高。茎粗、株高、茎数构成芦笋产量、品质特性,同时也是新品种选育的必备性状。通过对产量和一级笋率的多重比较发现,全雄品种在产量、一级笋率和其它农艺性状方面明显优于其它常规种,因此芦笋全雄品种的选育已成为目前芦笋育种的重中之重。