花楸合子胚发育的形态解剖学观察

为了解花楸有性生殖过程及败育现象,采用石蜡切片法对不同发育阶段的花楸合子胚进行了形态解剖学观察。结果表明:花楸属于核型胚乳;花楸受精后合子静止期为5 d左右。花楸合子(受精卵)发育经过原胚、球形胚、心形胚、鱼雷形胚、子叶胚等5个阶段。在5月21日的切片材料中首次观察到受精卵。开花后6～9d(6月初)发育成原胚,开花后约15 d发育成球形胚,之后5 d发育成心形胚,开花后1个月观察到鱼雷形胚,再经过10 d左右发育成子叶胚,7月15日前后合子胚基本发育成熟。在花楸子叶胚初期有较严重的败育现象。     

山茱萸胚和胚乳发育的初步研究

山茱萸胚乳的发育类型为核型,种子为有胚乳种子。胚的发育可分为三个时期:原胚期—分化期(快速生长期)—成熟期,胚的生长呈S型曲线。淀粉在山茱萸胚、胚乳中的积累有顺序性。     

中国鹅掌楸与北美鹅掌楸种间杂交的胚胎学研究*

应用控制授粉、常规石蜡切片技术研究了中国鹅掌楸与北美鹅掌楸种间杂交后的胚胎发育过程。结果表明：（１）控制授粉后花粉正常萌发，并有较多的花粉管进入柱头，但花粉管在花柱内生长缓慢甚至解体；（２）花粉萌发６ｄ后花粉管经花柱沟、珠孔塞和珠心冠原组织进入胚囊进行珠孔受精，２０ｄ时游离核胚乳细胞化，发育形成２～４个细胞厚的狭长组织，３５ｄ后可观察到球形胚；（３）随着胚和胚乳的发育，珠被发育形成种皮。授粉时，胚珠具２层珠被，外、内珠被均由外表皮、中层和内表皮组成；胚与胚乳发育期间，珠被各层发生了一系列的变化，最终种皮由外珠被的外表皮细胞形成的膜层和中层细胞形成的气室以及内、外珠被内表皮细胞形成的硬化层组成；（４）受精时成熟胚囊及到达珠孔的花粉管的数目较少，受精频率较低；（５）受精过程基本正常，但胚和胚乳发育不协调，导致胚在发育早期败育，很难观察到后期胚的发生。     

小叶杨(Populus simonii Carr.)种子发育的胚胎学观察

在人工授粉条件下,观察了小叶杨有性生殖过程的形态发育及其持续时间,结果如下:除正常花粉外,有异常和败育花粉,但不影响传粉和受精。胚珠倒生,单珠被,厚珠心,蓼型胚囊,珠孔受精,核型胚乳,胚的发育属柳叶菜型,种缨由胎座表皮细胞发育而成,柱头属湿型,花柱是实心的。人工授粉后4小时,花粉粒90％以上萌发,授粉后48小时,花粉管到达胚囊,双受精作用发生在授粉后3—6天,胚乳游离核时期持续约15—18天,但从胚乳游离核到胚乳细胞全部形成只需2—3天,合子“休眠”6—10天,原胚期约10天,心形期和鱼雷期各约2天,幼胚后期发育迅速,胚体由占种子内全长的1/3至充满整个种子只需5—6天。从授粉到种子成熟,历时约一个月。     

广西板栗胚珠形成与胚胎发育

以广西10年生地方板栗品种东兰油栗为研究对象,通过石蜡切片技术对板栗胚胎发育过程进行观察。结果表明:5月31日—6月7日,板栗雌花发育进入胚珠形成期,胚珠为倒生。到6月12日和6月21日时,大孢子母细胞经过减数分裂,发育成为具有4个成熟细胞核的胚囊和8个核的成熟胚囊。6月23日观察到板栗处于受精卵形成或分裂时期,同时伴有8~10胚乳核细胞围绕在受精卵周围,中央为大液泡。6月30日采集的样品中,胚体成球形均匀增长,其细胞分裂方式以平周、垂周分裂为主。7月8日为心形胚形成期到子叶发育期,已经能观察到形成完全的胚,有子叶细胞形成。胚乳细胞体积大,细胞壁薄,有明显的大液泡;7月15日可以观察到子叶细胞充满到整个子房内,胚乳细胞逐渐退化消失。     

墨西哥落羽杉×落羽杉杂交的生殖与发育解剖学特征

【目的】了解墨西哥落羽杉×落羽杉杂交的生殖与发育解剖学特征,为新品种、良种选育及种子发育的分子调控机制研究提供参考。【方法】通过形态观测和石蜡切片法,对杂交母本墨西哥落羽杉大孢子叶球从花芽分化至种子成熟的过程进行了系统的形态学和解剖学观察。【结果】1—5月为大孢子发生和雌配子体发育期。大孢子母细胞减数分裂形成4个大孢子,其中合点端1个发育为功能大孢子,其余3个退化。墨西哥落羽杉的复合颈卵器由13～19个颈卵器组成,顶生,无腹沟细胞,复合颈卵器外侧具1层双核套层细胞。6月中旬为受精期,随着花粉管的生长,精原细胞体积不断增大,并在受精前分裂为2个同型精子,其中1个与卵细胞受精形成合子,精卵融合主要发生在颈卵器中上部。6月下旬—7月上旬为原胚发育期,7月中旬—8月上旬为早期胚发育期,8月中旬—9月中旬为后期胚发育期。墨西哥落羽杉原胚发育属于松杉类标准型,简单多胚和裂生多胚现象并存,胚发育不同步,成熟胚为直线型,具4～9枚子叶,胚轴中无髓。【结论】本研究首次从细胞学水平上观察了墨西哥落羽杉大孢子发生和雌配子体发育过程,探究了墨西哥落羽杉×落羽杉生殖融合和胚胎发育类型,为墨西哥落羽杉×落羽杉的种...     

油料植物琴叶风吹楠种子萌发观察

琴叶风吹楠(Horsfieldia pandurifolia)幼苗属于子叶留土类型。成熟种子的主要成分是胚乳,胚极小,位于种子基部;胚根先突破位于种子基部的发芽孔形成主根,然后上胚轴开始生长,子叶向胚乳组织延伸以吸收营养物质;少数种子具多胚现象;只要播种方法正确,容易获得生长正常的苗木,否则容易导致苗木畸形。     

极度濒危植物水松大孢子发生、雌配子体发育及胚形成

【目的】通过对水松生殖发育解剖结构的研究,探讨水松属分类地位,分析水松种子严重败育的原因,进一步为水松繁殖和保护提供丰富、完善的胚胎学研究资料。【方法】2011年9月至2013年9月,采用石蜡切片技术对水松大孢子发生、雌配子体发育和胚形成过程进行系统地显微观察,详细观测水松雌球果发育过程中外部形态变化。【结果】9—11月为水松雌球果芽分化期,12月至翌年5月为大孢子发生和雌配子体发育期,6月为受精期,6月下旬至7月为原胚发育期,8月为早期胚发育期,9月为后期胚发育期。大孢子母细胞减数分裂形成直线型四分体,其中3个大孢子逐渐退化,仅靠近合点端的大孢子发育成为功能大孢子;复合颈卵器由6～17个颈卵器构成,外层有双核套层细胞。套层细胞具有明显的极性分布;复合颈卵器大多数着生于雌配子体顶端,少数侧生;中央细胞不形成卵核和腹沟细胞,直接发育为成熟的卵细胞;精细胞进入颈卵器时,不损伤颈细胞;受精作用主要发生在颈卵器中部,偶尔发生在颈卵器两端。精核与卵核融合过程中,精核的核仁发生解聚和复聚;合子转移到合点端后,进行有丝分裂形成原胚自由核;受精作用属于有丝分裂前配子融合类型。原胚发育属松杉类标准型,为简单多胚类型;原胚发育形成原胚柄,但不形成初生胚柄。受精作用过程中,存在严重授粉不足现象。【结论】基于雌配子游离核、颈卵器和颈细胞数目,颈卵器类型和着生位置,中央细胞及原胚发育等特征综合分析表明,水松属与台湾杉属、水杉属、柳杉属、落羽杉属亲缘关系密切,与杉木属亲缘关系较近。授粉不足可能是水松大量瘪粒种子形成的主要原因。     

马尾松胚胎发育的观察研究

在自然条件下，马尾松成熟花粉粒发育需6个月，授粉有效期在珠鳞展开的头5天，授粉后雌配子体游离核时期持续12个月，从游离核到细胞约需7天，颈卵器2－3个，受精作用发生在授粉后第13个月，原胚期约30天，幼胚期约40，成熟胚期约60天；从球花芽分化到种子成熟约需24个月，在形态发育的观察中，发现低温冻害会引起花粉在发育各阶段的形态异常及大量败育。     

花楸树体细胞胚与合子胚的发生发育

利用花楸树人工控制授粉后的未成熟合子胚进行体细胞胚的诱导培养,通过石蜡切片、扫描电镜和透射电镜对合子胚和体细胞胚的发生发育过程进行组织、细胞观察。结果表明:1)体细胞胚发生进程与合子胚相似,均经历了球形期、心形期、鱼雷形期,最终发育成成熟胚;2)直接发生和间接发生在愈伤组织表面的体细胞胚均为单细胞起源,间接发生在愈伤组织内部的体细胞胚存在单细胞起源,同时还存在可能的多细胞起源;3)子叶和愈伤组织表面发生的体细胞胚在发育早期均具明显的胚柄,鱼雷形胚期胚柄退化;4)愈伤组织内部发生的体细胞胚没有胚柄;5)与合子不同,胚性细胞的第1次分裂为均等分裂;6)多细胞原胚周围降解的细胞具有细胞程序性死亡的形态特征。     

金钱松胚胎发育

本文主要阐述金钱松传粉、受精和胚胎发育各期进行的时间和特点。4月上、中旬花粉成熟为传粉期。5月花粉在珠心上萌发,2精核移入花粉管,同时颈卵器形成和卵细胞逐渐成熟。6月受精,受精卵形成新细胞质,继而发育成为4、8、16细胞至球形原胚,胚柄延长。7月胚体增大,根端先分化,苗端后分化。10月胚成熟。金钱松为简单多胚,原胚为松型原胚。     

水曲柳体细胞胚与合子胚发生的细胞学研究

水曲柳(Fraxinus mandshurica)属木犀科(Oleaceae)白蜡树属,是我国东北重要珍贵硬阔树种之一,主要分布于小兴安岭、长白山、辽宁东部山地等地区, 以材质优良而著称.由于长期不合理的采伐利用,目前可利用的资源急剧减少,已被列为国家三级保护植物(傅立国,1992).进行水曲柳体细胞胚胎发生的研究,在资源保护、树种快繁和基因工程育种上有其重要的现实意义.     

马尾松种子发育过程中种仁贮藏物质的积累与变化

连续４ａ以不同发育阶段的马尾松种子为试材，研究其种仁贮藏物的积累过程及其组分含量的变化规律，结果表明，（１）种仁的干重占成熟球果干重的４．８％，其贮藏物质随种胚发育而不断变化，粗脂肪，粗蛋白和总糖的最终含量分别为４４．７６％，４２．５０％和７．１４％（其中淀粉和可溶性糖分别为４．２６％与２．８８％）。（２）酶蛋白的最大含量在原胚阶段初为２０．８３％，以后至幼胚阶段急剧减少到４．９％。（３）脂肪由４     

Potential Effects of Temperature on Early Reproductive Development and Progeny Performance in Picea abies (L.) Karst

In order to enhance cone production in Norway spruce [Picea abies (L.) Karst] seed orchards have been established in more southerly, warmer sites in Norway. This has led to concern and some evidence that seedlings obtained from parent trees grown at warmer sites may not be well adapted if planted in northern regions. Selective events during sexual reproduction in the warmer climate may select for traits not well adapted for cold climates. This study describes ovule and male gametophyte development, fertilization and proembryo and early embryo development under six different temperature regimens of warm or cold, warm-cold or cold-warm, and compares development in relation to temperature sums in degree days (dd) and calendar date. The outside treatment in a cold climate in Norway served as the control, and normal development and few ovule abortions and abnormalities were observed. In no treatments were abnormalities in male gametophyte development observed. In treatments in which trees were kept in the greenhouse throughout the study or moved during the study from the outside to inside the greenhouse, a higher incidence of megagametophyte, egg or embryo abortion was generally observed.     

马尾松球果和种子的生长发育及养分动态*

１９９０～１９９３年连续４ａ，对马尾松球果和种子的生长发育与养分动态，进行了比较系统的分析研究。结果表明：（１）马尾松球果系２年成熟，授粉（４月上旬）后当年为幼球果形成期；第２年受精前的６月至以后的９月中旬为球果迅速生长期，９月下旬至１０月为球果成熟期。（２）在２年生球果生长期间，球果干物质不断增加，而水分含量逐渐减少，１０月底干物质与水分的含量达到平衡点。（３）球果养分的含量，ｌ年生球果幼期Ｎ＞Ｋ＞Ｐ，三者变化趋势近乎一致，２年生球果含Ｎ量明显下降。种子内的Ｎ、Ｋ含量在７月中两者相当，此后Ｎ含量上升直至成熟期，到１０中Ｋ与Ｐ的含量相近。（４）胚胎发育进程中，球果和种子的体积与养分含量在原胚和幼胚阶段（７月至９月中旬）的变化最大，以后渐趋平缓。     

Somatic embryogenesis from vegetative shoot apices of mature trees of Pinus patula

Embryogenic cultures were initiated and established from apical shoots of mature trees of three genotypes of Pinus patula Scheide et Deppe. Factors affecting initiation, including cold pretreatment, basal medium composition, growth regulators and gelling agent concentration, and the effect of partial desiccation on somatic embryo maturation were investigated. Cold pretreatment of thick sections (0.5-1.0 mm) of apical shoots at 2 degrees C for 3 days on 0.3% activated charcoal induced white mucilaginous embryogenic callus on initiation medium. Subculture of this embryogenic callus on maintenance medium resulted in the formation of embryonal suspensor masses with proembryos. Partial desiccation (12-90 h) of embryogenic tissue at the proembryo stage of development, prior to transfer to maturation medium containing 9 g l(-1) Gellan gum, enhanced somatic embryo maturation and germinability. The frequency of maturation increased from 5.3 to 16.5% after 12 h of desiccation and from 16.5 to 73.8% after 24 h of desiccation, but longer periods of desiccation were ineffective.     

Metabolite profiling reveals clear metabolic changes during somatic embryo development of Norway spruce (Picea abies)

Progress on industrial-scale propagation of conifers by somatic embryogenesis has been hampered by the differences in developmental capabilities between cell lines, which are limiting the capture of genetic gains from breeding programs. In this study, we investigated the metabolic events occurring during somatic embryo development in Norway spruce to establish a better understanding of the fundamental metabolic events required for somatic embryo development. Three embryogenic cell lines of Norway spruce (Picea abies (L.) Karst) with different developmental capabilities were studied during somatic embryo development from proliferation of proembryogenic masses to mature somatic embryos. The three different cell lines displayed normal, aberrant and blocked somatic embryo development. Metabolite profiles from four development stages in each of the cell lines were obtained using combined gas chromatography-mass spectrometry. Multivariate discriminant analyses of the metabolic data revealed significant metabolites (P ≤ 0.05) for each development stage and transition. The results suggest that endogenous auxin and sugar signaling affects initial stages of somatic embryo development. Furthermore, the results highlight the importance of a timed stress response and the presence of stimulatory metabolites during late stages of embryo development.     

银杏种子后熟生理与内源激素变化的研究

研究银杏种子后熟期间的生理变化以及内源激素对种子休眠与萌发过程的调控作用.结果表明:银杏种胚分化不完全、发育不充分是种子休眠的主要原因,银杏种子的后熟过程实质上是种胚不断分化生长的过程.在此过程中,物质代谢、酶活性的变化均有利于种胚发育和种子发芽,种子内部经历了一系列生理生化变化去调整内源激素平衡、完成种胚发育.研究发现,内源激素在银杏种子休眠与萌发过程中起着重要的作用:GA1 3是银杏种胚后熟的关键物质;ZRs对银杏种子生长起重要的促进作用;GA1 3/ABA及ZRs/ABA相对比例的变化对种子发育、胚的后熟及种子萌发均有重要的影响.     

Identification and quantitative analysis of stage-specific carbohydrates in loblolly pine (Pinus taeda) zygotic embryo and female gametophyte tissues

Stage-specific analyses of starch and 18 sugars, including pentoses, hexoses, disaccharides, trisaccharides, oligosaccharides and sugar alcohols, were made throughout seed development for zygotic embryo and female gametophyte (FG) tissues of loblolly pine (Pinus taeda L.). Tissue was most often analyzed in triplicate from two open-pollinated families grown in different locations and sampled in different years. Carbohydrates were analyzed by enzymatic assay, high performance liquid chromatography or gas chromatography/mass spectrometry. For all carbohydrates quantified, peak concentrations were higher in embryo tissue than in FG tissue. Significant changes in starch and sugar concentrations occurred over time, with both seed collections showing similar trends in temporal changes. Although concentrations were not always similar, embryo and FG tissues generally showed similar patterns of change in starch and sugar concentrations over time. Total starch concentration was highest during early seed development and decreased as development progressed. The major sugars contributing to osmotic potential during early seed development were D-pinitol, sucrose, fructose and glucose. During mid-seed development, D-pinitol, sucrose, fructose, glucose, melibiose and raffinose provided major contributions to the osmotic environment. During late seed development, sucrose, raffinose, melibiose, stachyose and fructose were the major contributors to osmotic potential. These data suggest stage-specific media composition for each step in the somatic embryogenesis protocol.