小叶兜兰的种子发育和无菌萌发

为建立小叶兜兰的繁育技术体系,本研究通过无菌播种的方法,辅以TTc生活力测定等方法,比较了小叶兜兰种子在授粉后不同发育时期和培养条件下的萌发率,对小叶兜兰种胚的发育过程进行了显微观察,探讨种胚发育程度与萌发的关系。结果表明,小叶兜兰种胚的发育阶段对萌发的影响最大,授粉后255d的种子萌发率最高（90．71％）,该阶段种子仍呈白色但微干燥,种胚刚发育至球形胚阶段,胚柄尚存。1／4MS和1／2MS为小叶兜兰适宜的基本培养基,添加100mg·L^-1的土豆汁对小叶兜兰的无菌萌发有良好的促进作用。     

扇脉杓兰果实生长动态及胚胎发育过程观察

对授粉后不同发育阶段扇脉杓兰(Cypripedium japonicum Thunb.)果实的生长动态进行了观察和分析,并分别采用TrC法和常规石蜡切片法研究了种子生活力及其胚胎发育过程.观察结果湿示:扇脉杓兰果实形态成熟时间约为110 d,其中,授粉后0～20 d为第1次迅速生长期,授粉后20～30 d为第1次缓慢生长期,授粉后30～50 d为第2次迅速生长期,授粉后50～110 d为第2次缓慢生长期;果实纵径和横径的生长动态变化过程相似,但横径的生长动态曲线较纵径平缓,形态成熟时果实的纵径和横径分别为48.87和13.59 mm.成熟种子由内外2层种皮和球形胚构成,不具胚乳,内外种皮间具空气腔;败育种子只具有内种皮和外种皮而无种胚.胚胎发育类型为石竹型,种胚自受精形成合子到发育为成熟球形胚约需95 d.种胚发育时合子第1次不均衡横裂形成基细胞和顶细胞;基细胞发育为胚柄细胞,胚柄细胞高度液泡化,在胚胎发育的过程中不进行分裂并逐渐退化消失;顶细胞不参与胚柄形成,并且经过有丝分裂最终形成球形胚;内珠被在种子成熟时发育成为1层致密的紧贴胚体的内种皮.种胚纵径和横径的生长动态变化相似,成熟球形胚的纵径和横径分别为208.71和106.19 μm.扇脉杓兰种子生活力较高,有生活力的种子占56％.根据研究结果推测:自然状态下扇脉杓兰种子萌发率较低,可能与致密的种皮、种子中较小的胚体以及无胚乳导致的营养成分不足有关.     

珍稀濒危植物金丝李种子的休眠机理

珍稀濒危植物金丝李(Garcinia paucinervis)种子的萌发十分缓慢,探讨其休眠机理,可为该物种的种质资源保育与可持续利用提供理论依据。本文对金丝李种子种皮结构及其透水性,剔除部分种皮和胚乳后种子的萌发情况,胚乳和胚等粗提物的活性,储藏、层积和不同温度下种子萌发情况,萌发过程中内源激素含量等进行了研究。结果表明:金丝李种皮无栅栏细胞层,下表面的角质层较薄;种皮对种子的吸胀阻碍小;随着种孔端剔除种皮和胚乳程度的加深,金丝李种子的萌发进程逐渐延长,甚至降低其萌发率,种脐端削除处理对种子萌发影响不大;内果皮、种皮、胚乳和胚中可能存在抑制金丝李种子萌发和生长的内源抑制物;新鲜种子胚率达86.12%,低温层积后胚率无显著变化;低温层积处理延缓其萌发进程,对萌发率无显著影响,4℃低温层积是储藏金丝李种子的较好方法;种子萌发对温度敏感,在32℃培养下可打破种子休眠,萌发速度显著加快。种子萌发过程中ABA含量降低,GA与ABA、IAA与ABA的比值随种子萌发显著升高,萌发促进与抑制物比例逐渐趋于提高。因此,金丝李种子存在内源抑制物,同时缺乏萌发促进物质,导致生理休眠。该种子休眠特性使其幼苗生长能应对生境的季节变化,种群在风险环境中得以延续,避免大量幼苗竞争。植被破坏导致种子萌发阶段受阻是造成金丝李濒危的原因之一。     

四种石斛兰种胚发育进程研究

以玫瑰石斛、尖刀唇石斛、短棒石斛、兜唇石斛种子为材料,进行种胚非共生萌发研究,并对其种子形态和胚的发育进程进行了显微观察。结果表明:处于球形胚阶段的石斛兰种子,种胚吸胀后突破种皮,发育至吸收毛和芽生长点出现后,种胚形成原球体;种子萌芽后胚尚未成熟,只进入心形胚阶段。呈纺锤形种子的种皮两端形状不同,一端存在结点,呈弯曲状的尖形,另一端种皮呈收拢的圆口形。4种石斛兰种子,玫瑰石斛种子最长,为两端狭长的纺锤形;兜唇石斛种子最短,呈两端稍细的纺锤形。玫瑰石斛、短棒石斛、尖刀唇石斛种子胚培养需要5～10 d萌发;兜唇石斛种子和胚皆偏小,萌发需要30 d。石斛兰种胚和种皮吸水膨胀后,种胚向种皮的一端移动、脱出或种胚撕裂种皮中央后突破而出,形成裸胚。玫瑰石斛种子撕裂种皮后主要从种皮中央突破;短棒石斛、尖刀唇石斛、兜唇石斛部分种胚从种皮一端脱出,部分种胚则从中央撑破种皮脱出。充分膨胀、变绿后萌芽的裸胚,存在极性,顶部芽生长点萌动,下部出现成群散射状吸收毛。     

鹤顶兰种子萌发及原球茎增殖培养研究

以鹤顶兰成熟蒴果的种子为实验材料,研究影响种子非共生萌发的因素和种胚发育途径,并采用正交设计研究了原球茎增殖的影响因素。结果表明:冷藏影响种子的活力且其萌发率随冷藏时间延长而降低;0.5%NaClO溶液浸泡种子可提高萌发率,缩短初始萌发时间;种胚发育途径为种胚转绿后从种子侧面突破种皮而形成原球茎,随后分化出具根芽结构的完整植株;6-BA对原球茎增殖作用显著,原球茎增殖的最适培养基为1/2MS+3.0mg/L6-BA+1.0mg/L KT+1.0mg/L NAA,增殖倍数为6.67。     

带叶兜兰种子原地共生萌发及有效菌根真菌的分离与鉴定

为获得带叶兜兰(Paphiopedilum hirsutissimum)种子萌发的共生真菌,采用原地共生萌发技术获得了2株自然萌发的小幼苗,并分离和筛选出了有效的种子萌发共生菌——瘤菌根菌(Epulorhiza sp.)。为验证分离菌株对带叶兜兰种子萌发的有效性,将Phs34号菌株与带叶兜兰种子在灭菌后的原生境基质上进行室内共生萌发试验,结果表明,经过6周的培养,对照组没有观察到种子的萌发;接菌的种子胚明显膨大,突破种皮,形成原球茎,平均萌发率为(58.35±3.41)%。这表明分离得到的瘤菌根菌能促进带叶兜兰的种子萌发。     

濒危植物巴东木莲种子休眠与萌发特性的研究

巴东木莲(Manglietia patungensis)为我国特有种, 属国家重点保护植物。为找出其生殖环节中的致危因素, 作者对巴东木莲种子休眠与后熟过程中的形态和萌发特性进行了研究。结果表明, 巴东木莲种胚发育不完全可能是种子休眠的主要原因, 在其后熟过程中胚不断分化、发育成熟; 种皮具有较好的透性, 与休眠的关系不大; 种子不同部位均存在萌发抑制物, 胚乳中高含量的萌发抑制物是影响胚萌发的重要因素。内源激素ABA和IAA在巴东木莲种子休眠与萌发过程中起着重要作用, ABA是引起休眠的关键因素, IAA有助于种子的萌发, IAA/ABA相对含量的变化对种子的休眠和萌发产生重要影响。巴东木莲种子的休眠是由种子本身的形态和生理特点引起的综合休眠, 在4℃低温保湿条件下才能完成其形态和生理后熟过程, 而自然条件下, 巴东木莲种子成熟时正值秋季少雨, 很容易失水而不能完成其后熟过程而失去生活力, 这可能是导致该物种自然更新困难的重要原因。     

几种兰花种子无菌萌发及胚胎发育过程的几种途径

以虎头兰(黄色素花)×大雪兰、大雪兰×虎头兰(黄色素花)、虎头兰(黄色素花)×黄蝉兰、黄蝉兰×虎头兰(黄色素花)、冬凤兰、竹叶兰、窄唇蜘蛛兰、半柱毛兰、三褶虾脊兰和多花脆兰为材料,进行了杂交和自交种子无菌培养研究,并对胚胎发育进行了显微观察,结果表明:兰花种胚的发育途径有两条,一是种胚突破种皮后再转绿;二是种胚转绿后再突破种皮,且两条途径是同时进行的。种胚突破种皮的方式有3种:一是种子发育过程中,种皮慢慢消失,变成裸露的胚;二是种胚向种子的一端移动,最终突破种皮;三是种胚从种子的一侧突破种皮,形成裸胚。幼苗的发育途径有两条:一是小球体经圆球茎发育成小苗;二是小球体伸长成根茎后发育成小苗。     

珍稀濒危植物蒜头果种子萌发特征与幼苗类型的研究

以去除果皮后阴干的珍稀濒危植物蒜头果种子为材料,在温室大棚沙池内进行层积处理,从层积处理开始至子叶出土的不同萌发阶段,考察蒜头果种子发育形态、贮藏物质积累、酶活性以及幼苗类型等变化特征,初步探讨其种子休眠形成原因。结果显示：(1)蒜头果种子从解除休眠开始至萌发形成幼苗的过程约需195 d,其中幼胚的形态发育后熟约需75 d,随后30 d内是种子集中萌发的时期,其发芽率达到最高(53.33%);依据种胚发育形态的标志特征将此过程划分为7个阶段(S1～S7阶段):S1阶段种子未萌发,S2阶段种胚“露白”,S3阶段胚根突破种皮长超过1 cm, S4阶段下胚轴与胚根连接处形成弯钩结构,S5阶段“S”型胚形成及胚根前端膨大,S6阶段种子不仅具有膨大的胚根且已有侧根的分化,S7阶段子叶脱落,胚芽出土,真叶出现。(2)蒜头果种子在湿沙层积过程中,种胚胚长和胚率从S1阶段的(5.49±1.57)mm和(19.48±5.72)%分别增加至S6阶段的(67.92±2.94)mm和(240.75±15.29)%,胚率平均增加了12.4倍,显示蒜头果种子的胚需要经历后熟过程才能萌发,属于胚后熟型种子。(3)从...     

新疆短命植物小拟南芥(Arabidopsis pumila)种子萌发特性及其生态适应性

通过显微结构及不同处理条件下种子萌发率的观察,对早春短命植物小拟南芥(Arabidopsis pumila)种子萌发特性及影响因素进行了研究,并对其生态适应性进行了讨论。结果表明:(1)温度和光照变化对自然生境和温室收获种子的萌发率影响均不显著,说明此种群在前期萌发阶段对光、温不敏感;(2)自然生境中采收的小拟南芥种子萌发率显著低于温室收获种子,说明环境条件的变化对短命植物种子的发育具有重要作用,可显著改变种子的萌发行为;(3)赤霉素使自然生境收获种子胚活性增强从而对萌发有较大促进作用,可使萌发率增加50%以上;(4)对种皮进行各种机械损伤处理使得种皮松弛或透气,可以显著提高自然生境种子的萌发率(超过70%);(5)盐和干旱胁迫对种子萌发均具有明显的抑制作用,但复水后部分被抑制种子可重新萌发,显示盐和干旱胁迫可导致种子产生浅度休眠。结合小拟南芥自然生存环境及本研究的结果,显示其种子萌发特性与生境具有高度适应性。     

Asymbiotic Seed Germination and in vitro Seedling Rapid Development of Paphiopedilum spicerianum

In this paper, we studied the asymbiotic seed germination and seedling development of Paphiopedilum spicerianum, a wild plant with extremely small populations (PSESP) in China. By analogizing the influences of different media, seed maturity, pretreatments and lights on the seed germination, the results showed that the best germination condition is using the seeds at 270 days after pollination, pretreating with 1% NaOCl for 40 min, on the 1/4MS+10% coconut water medium under 24 hours darkness for 4 weeks. For seedling formation, 30g·L-1 Hyponex No 1 medium with 10mg·L-1 α naphthlene acetic acid, 05g·L-1 activated charcoal and 10% banana homogenate was the most effective. For seedling development, the same medium used for seedling formation with supplemented 10g·L-1 6 benzyladenine was most suitable.     

Embryo development in the lady's slipper orchid, Paphiopedilum delenatii, with emphasis on the ultrastructure of the suspensor

BACKGROUND AND AIMS: Owing to large-scale collecting, the lady's slipper orchid, Paphiopedilum delenatii, is under threat of extinction. Asymbiotic germination provides a useful way to re-establish plants in the wild and for commercial propagation. A detailed study of embryo development would provide information on subsequent germination events and aid in the propagation of the species. METHODS: Developing capsules were collected for histochemical and ultrastructural studies by using both light and transmission electron microscopy. KEY RESULTS: The suspensor of this species consists of three vacuolated cells. During the early globular stage of embryo development, structural differentiation occurs, revealing an abundance of smooth endoplasmic reticulum cisternae and wall ingrowths within the suspensor cells. These features are not present in cells of the embryo proper. Furthermore, the results of Nile red staining demonstrate that a cuticular layer is present only in the embryo proper, but absent from the suspensor. Cuticular material is also present in the inner walls of the seed coat, and persists through seed maturation. CONCLUSIONS: The morphological features of the transfer cell and the absence of cuticular material in the suspensor cell wall corroborate the hypothesis that the suspensor is the major nutrient uptake site for the developing embryo in the lady's slipper orchid. The absence of an endosperm and presence of cuticular material in the inner walls of the seed coat enclosing the embryo proper further support the notion that nutrient uptake by the embryo is confined to the micropylar end of the seed through the suspensor.     

兰科杓兰属（Cypripedium）植物种子非共生萌发研究进展

兰科杓兰属(Cypripedium)植物主要分布于东亚、北美等温带地区和亚热带山地。杓兰不仅具有极高的观赏价值,而且其经济价值和科研价值也越来越受到人们的重视。近年来对杓兰属植物人工繁殖的相关研究不断深入,主要集中于种子的非共生萌发等方面。本文对濒危兰科杓兰属植物进行了简要介绍,并就其种子非共生萌发研究从种子成熟度、预处理和有机添加物的作用、培养基的配置等方面进行综述,为目标杓兰种类的非共生萌发试验方案的制定奠定基础,将有助于温带/高山兰科植物保育研究的发展。     

Asymbiotic seed germination and in vitro seedling development of <Emphasis Type="Italic">Cyrtopodium punctatum</Emphasis>: a propagation protocol for an endangered Florida native orchid

Cyrtopodium punctatum Lindley is an endangered epiphytic orchid restricted in the United States to southern Florida. Due to its ornamental value, the species was extensively collected from the wild during the past 100 years. Today, only a few plants remain in protected areas. As part of a conservation plan for the species, procedures for asymbiotic seed germination were developed. Five asymbiotic orchid seed germination media (PhytoTechnology Orchid Seed Sowing Medium, Knudson C, Malmgren Modified Terrestrial Orchid Medium, Vacin &; Went Modified Orchid Medium, and ½-strengh Murashige &; Skoog) were examined for their effectiveness in promoting seed germination and protocorm development under a 16/8 h L/D photoperiod and dark (0/24 h L/D). The influence of photoperiod on growth and development was also examined. Seeds were germinated under a 16/8 h, 12/12 h, 8/16 h L/D photoperiod, at 25 ± 3°C and allowed to develop in vitro for 10 weeks. After 10 weeks, developing seedlings were transferred to Sigma Phytatrays and returned to their assigned photoperiod treatments for continued seedling development for an additional 15 weeks. Highest germination occurred in 0/24 h L/D on PhytoTechnology Orchid Seed Sowing Medium and seedlings displayed more advanced development when cultured under 16/8 h L/D photoperiod after 15 weeks in Phytatrays. Thirty-five week old seedlings potted in coconut husk growing medium exhibited 90% survival following 5 weeks acclimatization to greenhouse conditions. This asymbiotic seed germination protocol for C. punctatum will facilitate future reintroduction projects involving this endangered species.     

培养基与光照对沼兰种子非共生萌发的影响

近年来由于灭绝性采挖和生态环境的恶化,沼兰(Malaxis monophyllos)的自然资源遭到严重破坏,加上沼兰种子具有萌发缓慢和萌发率低的特性,已处于近危乃至易危的状态。为了繁殖和保护沼兰种质资源,利用添加2%蔗糖、3%活性炭和0.8%琼脂的Knudson C(KC)培养基,研究了萘乙酸(NAA)、6-苄基腺嘌呤(6-BA)、椰乳、香蕉泥、土豆泥和光照条件6种因素对沼兰种子非共生萌发的影响;同时通过光学显微镜和扫描电镜观察了种子的外部形态。结果显示,沼兰种皮细胞的平周壁方向结构平坦、没有纹络,但是沿着细胞垂周壁方向细胞壁较厚。通过正交试验设计和冗余分析,建立了沼兰种子非共生萌发的最佳方案,即在含有4.5mg·L–1NAA、10mg·L–16-BA、8%香蕉泥和3%土豆泥的KC培养基上光照条件(光照强度为20μmol·m–2·s–1,光周期为12h/12h)下萌发。该方案的种子萌发率在90%以上。     

Seed Germination and Seedling Development of Several Tree Species in the Lower Subtropical Evergreen Broad-Leaf Forest

Studies on seed germination and seedling development of Ormosia glaberrima Wu, Ouercus hui Chun, Diospyros morrisiana Hance, Oroxylum indicum (L.) Vent., Toona sinensis (A. Tuss. ) Roem. and Paulownia fortunei (Seem.) Hemsl. revealed that had rapid germination but Ormosia glaberrirna. Germination began 2--3 days and ceased 2—4 weeks after the initiation of the experiment. Fast-growing sun trees Oroxylum indicurn, T. sinensis, and P. fortunei germinated faster than did the species in the forest. Germination of Oroxylum glaberrirna was delayed because of its hard seed coat, but became rapid when its coat was slit open. Species in the forest and Oroxylurn indicurn and T. sinensis could germinate in both dark and light, tiny seeds of P. fortunei germinated poorly in dark. Water absorption of seeds presented three phases which varied among the species. Morphology of the seedlings might be related to shading-tolerance and growth speed of species.     

Asymbiotic germination response to photoperiod and nutritional media in six populations of Calopogon tuberosus var. tuberosus (Orchidaceae): evidence for ecotypic differentiation

Background and Aims

Ecotypic differentiation has been explored in numerous plant species, but has been largely ignored in the Orchidaceae. Applying a specific germination protocol for widespread seed sources may be unreliable due to inherent physiological or genetic differences in localized populations. It is crucial to determine whether ecotypic differentiation exists for restoration and conservation programmes. Calopogon tuberosus var. tuberosus, a widespread terrestrial orchid of eastern North America, is a model species to explore ecotypic differences in germination requirements, as this species occupies diverse habitats spanning a wide geographical range.

Methods

Mature seeds were collected from south Florida, north central Florida, three locations in South Carolina, and the upper Michigan peninsula. Effects of three photoperiods (8/16, 12/12, 16/8 h L/D) were examined on asymbiotic in vitro seed germination and seedling development of C. tuberosus. Germination and early development was monitored for 8 weeks, while advanced development was monitored for an additional 8 weeks. In an additional experiment, asymbiotic seed germination and development was monitored for 8 weeks on six culture media (BM-1 terrestrial orchid medium, Knudson C, Malmgrem, half-strength MS, P723, and Vacin and Went). A tetrazolium test for embryo viability was performed.

Key Results

Short days promoted the highest germination among Florida populations, but few differences among photoperiods in other seed sources existed. Different media had little effect on the germination of Michigan and Florida populations, but germination of South Carolina seeds was higher on media with higher calcium and magnesium. Tetrazolium testing confirmed that South Carolina seeds exhibited low viability while viability was higher in Florida seeds. Seed germination and corm formation was rapid in Michigan seeds across all treatments. Michigan seedlings allocated more biomass to corms compared with other seed sources.

Conclusions

Rapid germination and corm formation may be a survival mechanism in response to a compressed growing season in northern populations. Ecotypic differentiation may be occurring based on seed germination and corm formation data.Key words: Asymbiotic germination, corm development, Calopogon tuberosus, ecotypic differentiation, native orchid, orchid seed germination, seedling development     

In vitro reproduction of three Limodorum species (Orchidaceae): impacts of scarification methods and nitrogen sources on mature seed germination and seedling development

Limodorum species are hard-to-cultivate, germinating with difficulty or not germinating at all on a number of media. The present work was focused on the effects of seed sterilization-scarification methods and of different nitrogen sources on in vitro seed germination and seedling development of Limodorum abortivum, L. brulloi and L. trabutianum. Our results showed that sodium hypochlorite solution with a 1% concentration of available chlorine was ineffective for both sterilization and scarification of Limodorum seeds, while a 5% concentration was found to be an effective sterilizing and scarifying solution for L. trabutianum. The positive effect of organic nitrogen in promoting asymbiotic germination in many terrestrial orchids was confirmed by this study for L. trabutianum and L. brulloi, which showed significantly higher germination percentages on Basal Medium modified than on Knudson C, which contains inorganic nitrogen. This is the first report of in vitro asymbiotic germination of mature seeds of L. brulloi and L. trabutianum, especially successful for L. trabutianum that showed high germination percentages (>63%) and an active seedling development. These results may be useful for conservation purposes, since L. brulloi is a narrow Italian endemics and L. trabutianum is threatened with extinction both in Italy and in France.