亚美万代兰与多花兰种子发芽过程的扫描电镜观察

兰种子发芽之后形成原球茎与根状茎,前者以亚美万代兰,后者以多花兰为材料进行扫描电镜观察。多花兰种子发芽初期,与亚美万代兰相似,但分化子叶的能力较差,在暗培养下,只形成鳞片状叶,紧贴在根状茎的分生组织上。根状茎只有转入光培养才有茎叶分化。兰种子发芽时的毛状物,在电子显微镜下观察,其形态与根毛较接近。     

卡德丽亚兰种子非共生萌发及萌发过程中原球茎发育的细胞学研究

对卡德丽亚兰种子非共生萌发条件及萌发过程中原球茎发育进行了研究。结果表明,低离子浓度培养基(KC、RE)利于卡德丽亚兰种子萌发;适宜种类和浓度的激素(1.0mg/LBA和0.1mg/LNAA或1.0mg/LKT和0.1mg/LNAA)对种子萌发有良好的促进作用。细胞学研究表明:卡德丽亚兰种子萌发过程的本质是未分化胚细胞团通过细胞的不断分裂形成原球茎,原球茎出现极性,随之分化形成芽端分生区,最终形成实生苗。     

天麻种子萌发过程中与其共生真菌石斛小菇间的相互作用

天麻Gastrodiaelata种子与石斛小菇Mycenadendrobii的共生萌发试验表明,石斛小菇可与天麻共生,促进天麻种子发芽并形成原球茎。菌丝主要分布于原球茎的柄状细胞、外皮层细胞和内皮层细胞,在外皮层细胞中形成菌丝结,内皮层细胞中的菌丝则被消化。原球茎细胞中的菌丝均被电子透明物质和原球茎细胞质膜包围而与原球茎细胞质相隔离,菌丝进一步液泡化并最终被水解。含有衰败菌丝的原球茎细胞常被菌丝重新定殖。这一菌丝被消化及菌丝的重新定殖过程在整个原球茎发育过程中可不断重复发生。     

天麻种子萌发过程中与其共生真菌石斛小菇间的相互作用*

天麻Gastrodiaelata种子与石斛小菇Mycenadendrobii的共生萌发试验表明,石斛小菇可与天麻共生,促进天麻种子发芽并形成原球茎。菌丝主要分布于原球茎的柄状细胞、外皮层细胞和内皮层细胞,在外皮层细胞中形成菌丝结,内皮层细胞中的菌丝则被消化。原球茎细胞中的菌丝均被电子透明物质和原球茎细胞质膜包围而与原球茎细胞质相隔离,菌丝进一步液泡化并最终被水解。含有衰败菌丝的原球茎细胞常被菌丝重新定殖。这一菌丝被消化及菌丝的重新定殖过程在整个原球茎发育过程中可不断重复发生。     

天麻种子萌发过程中与其共生真菌石斛小菇间的相互作用

天麻Ｇａｓｔｒｏｄｉａ ｅｌａｔａ种子与石斛小菇Ｍｙｃｅｎａ ｄｅｎｄｒｏｂｉｉ的共生萌发试验表明,石斛小菇可与天麻共生,促进天麻种子发芽并形成原球茎。菌丝主要分布于原球茎的柄状细胞、外皮层细胞和内皮层细胞,在外皮层细胞中形成菌丝结,内皮层细胞中的菌丝则被消化。原球茎细胞中的菌丝均被电子透明物质和原球茎细胞质膜包围而与球茎细胞质相隔离,菌丝进一步液泡化并最终被水解。含有衰败菌丝的原球茎细胞常被菌丝     

黑节草未成熟种子的形态发育及其在离体培养时的表现

黑节草(Dendrobium candidum Wall ex Lind.)2—6个月种龄的胚均处于球形胚阶段,不同种龄的胚在体积大小、胚细胞数目、胚细胞内的淀粉粒含量和超微结构上有差异。在离体培养条件下黑节草种子萌发率可达95％,种子萌发后形成原球茎,原球茎可以直接发育形成幼苗,又可以由原球茎产生大量愈伤组织,由愈伤组织再分化发育成幼苗。种子萌发过程中,胚顶端分生组织细胞的淀粉逐渐消耗,淀粉的变化与分生组织和子叶的形成有明显的相关性。     

大花蕙兰类原球茎形态发生的组织学研究

以大花蕙兰类原球茎发生发育不同阶段的培养物为试材,采用石蜡切片法对其进行组织学研究。结果表明,类原球茎形成过程中,表层薄壁细胞首先经脱分化恢复分生能力,形成愈伤组织,随后在其表面形成瘤状突起--类原球茎;类原球茎继续发育,在其周围形成多个不规则突起,并进一步发育形成新的类原球茎;而原来的类原球茎开始进入器官分化阶段,逐渐形成完整的小植株。根据观察,类原球茎起源可能存在两条途径:体细胞胚胎发生和器官发生。     

不同培养基上白芨的种子萌发与幼苗形态发生

对无菌条件下白芨种子在不同培养基上的萌发及其幼苗形态发生进行了研究.结果表明:在白芨种子萌发过程中,种胚先转绿,然后在种胚远离残余胚柄的一端突破种皮,形成原球茎.在原球茎顶端分化出叶原基,并逐渐发育成叶片.在原球茎下部表面存在透明的毛状物,推测这些毛状物与幼根根毛是同功的.白芨种子萌发最适培养基为1/2 g·L-1 +1.0 mMS NAA.添加低浓度的外源细胞分裂素(6-BA或KT)与NAA组合均严重抑制种子的萌发,种子萌发率较低,形成的圆球茎较小,原球茎分化出的小苗也较细弱.白芨生根壮苗最适培养基为1/2 g·L-1 NAA+2.0 g·L-1活性炭+1.0 mg·L-1 +1.0 mMS GA3.生根壮苗培养基中加入活性炭和GA3有利于根的生长和壮苗,且植株长势健壮.炼苗基质为腐殖土和蛭石(1∶1),成活率可达90%以上.     

同色兜兰的非共生萌发与快速繁殖研究

以授粉后不同发育时期的同色兜兰种子为材料,观察其形态特征和萌发过程,并探讨建立同色兜兰高效快繁体系的最佳条件。结果表明,种子发育中后期即授粉后210~240d为较适宜的采收期,授粉后210d的种子萌发率最高(达77.79%);1/4 MS和1/2MS为同色兜兰适宜的基本培养基,添加100mL/L椰乳或1g/L蛋白胨对种子萌发及原球茎生长和分化有明显的促进作用;添加1g/L活性炭对原球茎褐化有一定的抑制作用,但添加剂量不宜过大;添加香蕉汁和苹果汁对同色兜兰种子萌发和原球茎生长分化有抑制作用;暗处理对同色兜兰种子萌发无影响;分化后的原球茎在壮苗和生根培养基上培养120d即可得到4~5片叶、高3~5cm的同色兜兰健壮试管苗。     

濒危莲瓣兰杂交育种及原生地种子萌发的研究

以莲瓣兰'大雪素'(Cymbidium tortisepalum 'Daxuesu')和'剑阳蝶'(Cymbidium tortisepalum 'Jianyangdie')为亲本,研究莲瓣兰种间杂交育种、杂交种子在原生地播种共生萌发及植株形态发生过程,为莲瓣兰杂交育种新品种选育及杂交种子原生地播种种苗快速繁殖奠定基础.结果表明,莲瓣兰种间杂交结实率在90.0%～93.3%之间,杂交亲和性较强;莲瓣兰杂交种子在原生地播种6～12月后,能被萌发菌侵染、种胚突破种皮形成类原球茎,种子在原生地播种容易萌发;类原球茎转绿成根状茎、根状茎伸长成丛生型根状茎、丛生型根状茎顶端分生组织分化出原球茎、原球茎分化出叶和根、根状茎逐渐退化完成植株再生过程极其缓慢,需要4～6年的时间.     

Morphogenesis of the protocorm ofCypripedium acaule (Orchidaceae)

The nature of the protocorm of theOrchidaceae has fascinated morphologists for more than a century. In the present study, the development of the protocorm was followed using in vitro germination of seeds on a culture medium containing sugar, but without a symbiont. Inside the seed, the embryo consists of about a hundred cells. In the embryo, cells are arranged along a longitudinal axis according to size; these cells contain protein and lipid reserve material. In the first stages of seedling development, the embryo is transformed into a protocorm and meristematic tissue becomes organized into a meristematic dome (promeristem) at the anterior pole. This meristematic dome will give rise to a scale and the apex of the seedling. At first, the apex and the scale leaf develop synchronously. The development of the root always follows that of the apex. The study of the development of the seed ofCypripedium acaule showed that the protocorm is a distinct morphological system with respect to the rest of the cormus. The protocorm may be interpreted as an extension of the proembryonic stage.     

实验室条件下五唇兰菌根真菌专一性研究

利用从高原温带兰科植物菌根中获得的22个菌根真菌菌株, 对五唇兰(Doritis pulcherrima)进行了室内种子萌发、原球茎分化和组培苗回接试验, 从交叉回接的角度对附生兰科植物与菌根真菌的生理专一性进行了探讨。经过20周的共生培养, 只有编号为Cf1和Mm1的两个菌株使种子表现出种胚明显膨大的萌发迹象; 9个菌株能够促使原球茎较好地分化发育出根叶; 11个菌株处理苗的平均鲜重增长率高于对照组(156.25%), 其中Mm1的效果达到极显著水平(p = 0.01)。通过根切片显微观察, 在原球茎分化根和回接效果良好的处理苗的根皮层组织发现典型的菌丝团结构, 表明菌根体系已成功建立。温带地生兰菌根真菌对五唇兰种子萌发、原球茎发育和幼苗生长等3个重要生长阶段影响的试验显示, 五唇兰的种子和菌根真菌的共生萌发效果不佳, 而原球茎及幼株更容易与之建立良好的共生关系。同时, 也没有发现同一个真菌菌株能够对五唇兰的种子、原球茎和幼苗均产生促进作用。研究结果表明, 五唇兰的菌根真菌专一性因生理生长阶段的不同而存在差异。     

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。     

DNA synthesis and mRNA accumulation during germination of embryos of the orchidSpathoglottis plicata

Summary Germination of embryos of the orchid,Spathoglottis plicata Blume involves the formation of a protocorm in which DNA synthesis and cell divisions are confined to the proximal end whereas cells at the distal end undergo enlargement. Although³H-thymidine was incorporated into the distal cells of the embryo during the early period of germination, DNA synthesis was not followed by mitosis and cytokinesis. Poly(A)-RNA detected by in situ hybridization of sections to³H-poly-(U) was present uniformly in all cells of the embryo of the dry seed. However, coincident with the formation of the apical meristem, there was an increase in the density of auto-radiographic silver grains in the cells of the embryo, with a concentration of silver grains in the proximal part. The results indicate that regulatory events in the embryo prior to seed maturity determine the fate of its proximal and distal parts during germination.Dedicated to the memory of Professor John G. Torrey     

Endophytic fungi from Pecteilis susannae (L.) Rafin (Orchidaceae), a threatened terrestrial orchid in Thailand

Eight endophytic fungi were isolated from roots of the threatened terrestrial orchid, Pecteilis susannae (L.) Rafin. Phylogenetic analysis based on an alignment of internal transcribed spacer regions of nuclear rDNA indicated that seven isolates belonged to the genus Epulorhiza and one to Fusarium. All fungal isolates were cultured with orchid seeds collected from three field sites near Doi Suthep-Pui National Park, Chiang Mai, Thailand. Seed germination and protocorm development were evaluated up to 70 days after sowing. Percent symbiotic seed germination was highest (86.2%) when seeds were cultured with Epulorhiza (CMU-Aug 013). The protocorm development was the most advanced up to stage 2, continued embryo enlargement, or rupture of the testa, and the highest percentage was 17.8% when seeds were cultured with Epulorhiza (CMU-Aug 007). Without fungi, seed germination and protocorm development were 62.1% and 11.1%, respectively. The dependency of P. susannae on fungal symbionts for early seedling development is yet to be determined. Optimizing seed germination and seedling fitness will assist the conservation of this threatened orchid in Thailand.