天麻种子与兰小菇共生萌发过程中超微结构变化研究

天麻Ｇａｓｔｒｏｄｉａｅｌａｔａ种子与兰小菇Ｍｙｃｅｎａｏｒｃｈｉｄｉｃｏｌａ的共生萌发试验表明兰小菇可与天麻种子共生促进天麻种子萌发并形成原球茎。菌丝自胚柄端的柄状细胞侵入天麻种子原胚,其分布被限制在天麻原球茎基部的柄状细胞、外皮层细胞和内皮层细胞内,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。菌丝在外皮层细胞中形成菌丝结,在内皮层细胞中则被消化,形成扁化、衰败的菌丝或菌丝四块。含有衰败菌丝的原球茎细胞可被菌丝重新定殖,新近定殖的菌丝又被原球茎细胞消化。     

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     

天麻种子萌发过程与开唇兰小菇的相互作用*

实验表明开唇兰小菇Mycena anoectochila可与天麻Gastrodia elata种子共生促进其萌发形成原球茎。 菌丝自胚柄端的柄状细胞侵入天麻种子原胚,进一步在皮层细胞中扩展,在外皮层细胞中形成发育良好的菌丝结,菌丝完整而有活力; 在内皮层细胞中则被消化,菌丝衰败、扁化。菌丝在原球茎细胞内的分布被限制在原球茎基部的柄状细胞、外皮层细胞和内皮层细胞,菌丝均被电子透明物质包围, 外围环绕有原球茎细胞质膜, 该界面使侵入的菌丝与原球茎细胞质相隔离,也是两共生生物间进行物质交换的所在。上述菌丝侵入至被消化的过程在整个原球茎发育过程中可反复进行。     

天麻种子萌发过程与开唇兰小菇的相互作用

实验表明开唇兰小菇Mycena anoectochila可与天麻Gastrodia elata种子共生促进其萌发形成原球茎,菌丝自胚柄端的柄状细胞侵入天麻种子原胚,进一步在皮层细胞中扩展,在外皮层细胞中形成发育良好的菌丝结,菌丝完整而有活力;在内皮层细胞中则被消化,菌丝衰败,扁化,菌丝在原球茎细胞内的分布被限制在原球茎基部的柄状细胞,外皮层细胞和内皮层细胞,菌丝均被电子透明物质包围,外围环绕有原球茎细胞质膜,该界面使侵入的菌丝与原球茎细胞质相隔离,也是两共生生物间进行物质交换的所在,上述菌丝仞入至被消化的过程在整个原球茎发育过程中可反应进行。     

天麻种子／原球茎和营养繁殖茎被菌根真菌定殖后的细胞分化

天麻（Gastrodia elata Bl.)种子可与紫萁小菇（Mycena osmundicola Lange),兰小菇（M.orchidicola Fan et Guo)等一类小菇属真菌共生萌发形成原球茎。侵入种皮的菌丝集结在柄状细胞外周的胚柄残迹中,首先侵入胚的柄状细胞,然后自柄状细胞侵入其他原胚细胞。原胚细胞发生功能分化,形成菌丝结细胞和消化细胞,初被菌丝定殖的原胚细胞具有消化菌丝的功能,随后,部分原胚细胞逐渐被菌丝充满,充育成菌丝结细胞。菌丝由菌丝结细胞进一步侵入消化细胞后最终被消化。由原球茎分化形成的营养繁殖茎（以下简称营繁茎）进一步被蜜环菌（Armilariella mellea(Vahl.Fr.)Karst.)定植,蜜环菌与紫箕小菇的菌丝同时存在于营繁茎中,但两菌相遇时都停止蔓延,互不交错侵染。     

天麻消化紫萁小菇及蜜环菌过程中细胞超微结构变化的研究

本文对天麻种子消化入侵的紫萁小菇菌丝及营养繁殖茎消化蜜环菌过程中,细胞超微结构的变化进行了研究。观察结果表明:紫萁小菇侵入种胚后,染菌胚细胞的细胞器逐渐消失,其细胞质产生囊状体起消化菌丝的作用,存在于胚细胞中的紫萁小菇菌丝有脱壁或失去细胞质成为空腔等变化;种子萌发形成的原球茎消化紫萁小菇的方式同胚萌发阶段类同。蜜环菌侵入原球茎分化的营养繁殖茎后,皮层细胞产生消化酶类颗粒或囊状体包围并分解蜜环菌菌丝;被皮层细胞消化的菌丝残物或部分菌丝进入皮层内面的大型细胞,此时大型细胞的代谢功能显著增强,该细胞中的各种水解酶颗粒及液泡等完成对菌体物质的最终同化。紫萁小菇及蜜环菌先后在天麻有性繁殖和无性繁殖阶段侵染供给其营养,但菌丝被消化过程中的细胞形态变化、被消化方式不完全一致。     

应用放射性自显影技术研究标记紫萁小菇侵染天麻种胚的过程

用~3H—葡萄糖对紫萁小菇进行氚标记,然后用标记的紫萁小菇伴播天麻种子,发现标记紫萁小菇在侵入天麻胚柄状细胞后,菌丝即被溶解、消化,未被侵染的大型细胞中有标记化合物的踪迹。随着胚的发育,在胚顶端分生细胞中也有大量标记物的显影银粒云集,表明同化的标记化合物由细胞壁进入并通过胞间连丝进行运输,直观证明了紫萁小菇为天麻种子萌发提供了营养。     

墨兰菌根的结构及酸性磷酸酶定位研究

利用光学显微镜、电子显微镜及细胞化学方法,对墨兰菌根的结构和酸性磷酸酶定位进行了初步研究。结果表明墨兰具有典型的兰科植物根结构,发现该兰花的根的外皮层不具薄壁通道细胞,菌根真菌通过破坏部分根被和外皮层细胞而侵入根的皮层细胞并在细胞内形成菌丝结,侵入的菌丝被染菌皮层细胞质膜和电子透明物质包围,进一步被消化并聚集成衰败菌丝团块。酸性磷酸酶在染菌皮层细胞及包围菌丝的皮层细胞质膜和衰败菌丝细胞壁上有强烈的酶反应,衰败菌丝周围分布有许多单层膜的含酶小泡,它们可相互愈合形成大的含酶泡或与包围菌丝的质膜融合,类似于兰科植物共生原球茎中观察到的现象。说明皮层细胞可主动释放水解酶参与对菌丝的消化     

Cytodifferentiation of the Seeds (Protocorms) and Vegetative Propagation Corms Colonized by Mycorrhizal Fungi

The stipecell, subepidermal parenchyma cells and inner cortical parenchyma cells were differentiated from Gastrodia elata Bl. seed and protocorm cells when they were colonized by the fungal hyphae of Mycena osmundicola Lange and M. orchidicola Fan et Guo. The hyphae aggregated in the suspensor remnant surrounding stipecell, primarily penetrated the stipecell, and then colonized the embryo of seed. Stipecell is the unique invading site of the hyphae. Subepidermal parenchyma cells containing pelotons of hyphae is also a kind of passage cells of hyphae, but, when primarily colonized by hyphae, they can degenerate a little of hyphae. The hyphae colonizing inner cortical parenchyma cells were totally degenerated, and the function of inner ocrtical parenchyma cells is digestive. The vegetative propagation corms, which differentiated from protocorms, were recolonized by Armilariella mellea (Vahl:Fr.) Karst., and the hyphae of A. mellea and M. osmundicola were found in the same cell, but there is a layer of cells uncolonized by mycorrhizal fungal hyphae. This means the two fungal species can not crisscross colonize the cell of G. elata.     

Cytological studies on rust fungi. (vi) fine structures of infection process of Kuehneola japonica (diet.) dietel

The behavior of rust fungi in their host plants has been elucidated by electron microscopy. However, most of the ultrastructural studies on rust fungi have focused on the uredial stage. In order to elucidate the features of the sporidial stage, we studied the fine structure of Kuehneola japonica, a short-cycle rust, in rose leaves. Infection pegs arising from appressoria penetrated the host walls. Papillae formed at the time of penetration against the outer epidermal cell walls. The papillae which had formed at the penetration sites grew extensively and partially surrounded the intracellular hyphae which were connected with the infection pegs. The intracellular hyphae in the epidermal cells developed further and entered adjacent parenchyma cells. Walls of parenchyma cells either invaginated or thin papillae formed at penetration sites and the invaginated walls or papillae surrounded the necks of the intracellular hyphae. Intracellular hyphae in both epidermal and parenchyma cells were not enveloped by the sheath before 20 days after inoculation. In specimens prepared 20 days after inoculation, some of the intracellular hyphae were enveloped by a sheath in both palisade and spongy parenchyma cells. The sheathed hyphae resembled haustoria of other rust fungi which had been described previously. Teliospore initials were formed in mycelial masses in intercellular spaces between the epidermal cells and palisade parenchyma cells 20 days after inoculation. Uninucleate teliospores developed from teliospore initials 30 days after inoculation.Contribution No. 32.     

墨兰试管苗植株的发育解剖学研究

用石蜡切片和扫描电镜对墨兰试管苗植株的生长发育进行了研究。发现幼叶中脉附近的叶肉细胞类似栅栏组织,随着叶片的不断成熟,叶片基部中脉附近的叶肉细胞逐渐变为近圆形或椭圆形,而叶尖部和叶中部中脉附近的叶肉细胞仍似栅栏组织。茎的发育经历了原球茎、根状茎和假鳞茎3个阶段。原球茎的大部分细胞都含有淀粉粒,根状茎的皮层细胞含淀粉粒,而假鳞茎几乎不含淀粉粒。幼根没有髓,皮层细胞含淀粉粒：成熟根具含淀粉粒的髓。出瓶苗上即带有4个芽,一般只有最外侧叶腋的1个花芽和最内侧叶腋的1个叶芽发育。     

Association between microtubules and symbiotic fungal hyphae in protocorm cells of the orchid species, Spiranthes sinensis

Seeds of the orchid species, Spiranthes sinensis (Pers.) Ames, were sterilized and germinated in vitro with the symbiotic fungus Ceratobasidium cornigerum (Bourdot) Rogers. Colonized embryos developed into protocorms and these were examined for changes in microtubule arrays, after initial invasion of fungal hyphae into embryos and during peloton formation and degradation. Methods utilized to detect microtubules included immunofluorescence combined with laser scanning confocal microscopy, conventional transmission electron microscopy combined with morphometric analysis, and immunogold labelling. Microtubules were regularly found in close association with intracellular hyphae and degraded hyphal masses. Cortical microtubules disappear during peloton formation but reappear in cells that show fungal lysis. With conventional transmission electron microscopy and immunogold labelling the microtubules associated with fungal hyphae and degenerated hyphal masses were located close to the perifungal membrane that separates fungal hyphae from protocorm cytoplasm.     

Cytological Study of Wheat Spike Infection by Bipolaris sorokiniana

The infection of wheat spikelets by Bipolaris sorokiniana , the causal agent of black point on grains and grain shrivelling, was examined by light and electron microscopy. Conidia of the pathogen germinated 6–12 h after inoculation on the surfaces of the different spike tissues. Extracellular sheaths were observed on germ tubes and appressoria attached to the surfaces of lemma, palea and seeds, but were only scarcely detected on the surface of conidia. Appressoria, frequently found over grooves, formed penetration hyphae invading the epidermal cell walls. Infection process was similar on the surface of the lemma, palea and glume. Growth of the fungus in the epidermal and parenchyma cells was found predominantly in the cell walls, and hyphae also extended intercellularly and intracellularly. Infection of seeds appeared to occur via two ways: (i) direct infection of the outer layers of the cell walls of the pericarp and (ii) through entering the stigma into the pericarp cells. Secretion of host cell wall hydrolytic enzymes at the apex of the penetrating hyphae may facilitate the spread of the fungus. In addition, toxins secreted by the fungus might explain the rapid death of host cells in contact with or distant to fungal cells. A host response to fungal infection involved the development of appositions between cell wall and plasma membrane in cells adjacent to fungal cells. Fungal hyphae were sometimes also surrounded by electron dense material.