蜜环菌侵染后猪苓菌核防御结构的发生及功能

蜜环菌通过猪苓菌核表皮侵染菌核时,菌核表皮下层的菌丝具特异的拮抗反应,如细胞中有结晶颗粒出现,厚壁菌丝形成,部分薄壁菌丝有质壁分离现象。蜜环菌的侵染诱导了菌核防御结构的发生:离侵染点一定距离的部位出现由少量木质化菌丝和厚壁菌丝形成的疏松带状;蜜环菌侵入后,上述菌丝增多并紧密排列形成菌核的初级隔离腔,入侵的蜜环菌和部分菌核被隔离在腔中;在初级隔离腔形成的同时,外围的次级隔离腔开始发育。蜜环菌菌索和皮层菌丝分枝可突破初、次级隔离腔的壁,再以菌索产生的侵染带侵染菌核较外部的最后防线即三级隔离腔。本文较系统的阐述了蜜环菌侵染后菌核各防御结构的发生特点及功能。     

猪苓与蜜环菌的关系

蜜环菌Armilla riella mcllea(Vahl：Ff．)Kgtst·侵入猪苓Gri[ola umbe』zd‘4(PetsFr．)PilOt)菌核,激活了猪苓菌抵御异体侵染免疫反应的本能,猪苓菌丝细胞木质化,形成与菌核表皮结构相似的隔离腔,将蜜环菌素和部分猪苓菌丝包围。在隔离腔中蜜环菌消化分隔在腔中的猪苓菌丝,另外猪苓菌丝也可侵入或附着在蜜环菌索及侵染带细胞间隙吸收其代谢产物,猪苓菌核即可萌发出新苓正常生长。当隔离腔中的猪苓菌丝被消化后,蛮环菌生活力也减弱,解体后被猪苓菌吸收利用,隔离腔变成空腔。从广义角度看,仍可把蜜环菌与猪苓菌寄生与反寄生的营养关系概括为共生关系。     

Nutrient Source of Sclerotia of Grifola umbellata and Its Relationship to Armillaria mellea

Ways nutrient-uptake of sclerotia of Grifola umbellata and the relationship between G. umbellata and Armillaria mellea were studied. At the primary stage of sclerotia of G. umbellate infected by A. mellea, the hyphae of G. umbellata could obtain nutrients by invading the one- to three-layer-cells of A. mellea cortex which existed in the sclerotia of G. umbellata, ar late stage of A. mellea infection, the nutrient source of sclerotia of G. umbellata mainly depended on its hyphae, adhering on the intercellular space of A. mellea, to suck the metabolic products of A. mellea. After being nourished the hyphae of sclerotia of G. umbellata outside of the rhizomorph of A. mellea began to reproduce, as their nuclear divisions were well observed. The results suggested that the mutual assimilation between G. umbellata and A. meIlea could be defined as a form of special Symbiotic relation.     

猪苓、伴生菌及蜜环菌共培养的形态学研究

本文对猪苓、伴生菌及蜜环菌两两共培养及三者共培养进行了宏观形态观察及细胞学水平上的研究。结果表明,猪苓与伴生菌共培养时,在二者之间形成一致密拮抗线,猪苓菌落表面菌丝分化产生大量菌丝束;猪苓与蜜环菌共培养时,猪苓能阻止蜜环菌菌索对其自身的进一步侵袭,互作区中的双方菌丝及菌索均停止生长;蜜环菌与伴生菌共培养时,蜜环菌能穿透整个伴生菌菌落,在伴生菌菌落下方产生大量分枝;三者共培养后,猪苓对蜜环菌的防御能力有所下降,伴生菌对蜜环菌的耐受力有所提高,蜜环菌产生的新分枝均向伴生菌一侧生长,猪苓与伴生菌之间并不形成致密拮抗线,只可见双方菌丝的白色交融区。 猪苓与伴生菌均能在蜜环菌菌索皮层上形成侵入位点。     

蜜环菌索发育的研究

利用光学和电子显徽镜对蜜环菌索的发育及其结构分化进行了较系统研究。菌索的顶端有保持细胞不断分裂的分生组织区。由此衍生的菌丝细胞组成菌索的初生结构,包括分化不明显的表皮、皮层及初生髓;初生髓细胞体积大,核同步分裂产生多核体细胞,以一个或几个核为单位在爵体细胞中分化出细长的菌丝后,可以出芽方式自母体细胞中伸出,并且一开始就有薄壁与厚壁之分,同一母体细胞中可同时产生这两类菌丝。发育后期母体细胞破裂形成菌索的髓,两类疏松菌丝分布在其中。观察了成熟菌索的结构和侧枝的形成过程。菌索侧枝起源于皮层细胞,该细胞横向分裂首先形成分枝原基,之后突破菌索壳而分化出新的菌索顶端。讨论了蜜环菌索在不同寄主中的侵染方式。     

猪苓菌丝形成菌核伴生菌的发现及应用

在野生猪苓(Grifola umbellata (Pers.) Pilát)菌核生长穴中首次分离到猪苓菌丝形成菌核所必需的伴生菌(Grifola sp.).实验证明:纯培养的猪苓菌丝不能形成菌核,但其与伴生菌共培养,无论在实验室培养基上或用树棒栽培,猪苓菌核形成很快且发育正常;伴生菌是猪苓菌丝形成菌核的关键生物因子.另外,伴生菌菌丝和猪苓菌菌丝二者形态差别较大,前者多为细长的薄壁菌丝,后者多为细胞直径较大的薄壁和厚壁菌丝.     

猪苓菌核结晶及厚壁细胞的起源与发育

猪苓菌核的含晶细胞发生于菌丝中间或顶端,该细胞具有体积大、细胞质丰富等特点;结晶是由细胞质中的微小颗粒沉积于液泡中逐渐发育而成,液泡周围常有数量较多的线粒体分布,结晶发育至一定大小时细胞壁破裂释放出结晶,单个结晶在菌核中可聚集成大的棱状晶体。厚壁细胞产生于菌丝中间,与两端细胞以横隔膜相隔,细胞质收缩的同时胞壁加厚,厚壁细胞发育至仅留很小胞腔或完全被加厚物质充满时,可与相邻菌丝细胞分离;猪苓菌核厚壁细胞与有些真菌无性厚壁孢子的形成类同,但其大小不等在5～30μm之间。     

Role of Lysosomal Vesicles in the Digestive Process of Armillaria mellea by the Large Cells of Gastrodia elata

The hyphae of Armillaria mellea Fr. invade the large ceils of Gastrodia elata BI. Through the wall pits of cortical cells. During early stage the plasmalemma of large cell invaginates and the cell wall forms papillary thickenings to restrain the hyphae from invading. When a hypha enters a large cell, it is encircled tightly by the invaginated plasmalemma which is surrounded by a large number of vesicles coated by a unit membrane. As these vesicles fusing with their membranes to the plasmalemma and discharging their contents into the space around the hypha, the space lined by the invaginated plasmalemma enlarges gradually and becomes a digestive vacuole in which a hypha is completely digested. Reaction product form acid phosphatase activities in the vesicles and digestive vacuoles testifies that the vesicles and digestive vacuoles are identical with primary and secondary lysosomes of plant lysosomal system respectively.     

Studies on the Character istic of the Structures in the Cortical Cells of Gastrodia clara after Infection of Armillaria mellea

Aided by the techniques of thin and ultrathin sectioning and electron microscopy, the characteristic of structures in the cortical cells of Gastrodia elata was further investigated after infection of Armillaria mellea. It was found that the “hyphal coils”, observed with light microscope, in the cortical cells of G. elata were saccate structures deriv- ed from the cytoplasm of cortical cells and enclosed hyphae. And the cell walls of hyphae were digested in these sacs. Then, these hyphae without cell wall were cut into protoplast fragments in inner-most cortical cells. The results indicated that the cortical cells of G. elata possess digestive function.     

The Biological Relationship of Gastrodia elata and Armillaria Melle

We have done a series of experiments on the biological relationship of G. elata and Armillaria mellea (Vahl ex Fr.) Quel on the basis of popular experience. The results obtained are as follows: 1. The whole ontogenetic pattern of G. elata may be generally divided into four stages: seedling formation, tuber formation, flowering and fruiting. Except for the flowering and fruiting stage, the activity of Amella determines the formation of G. clara seedlings, the renewal of its vegetative organs, and transformation of vegetative growth to reproductive growth. The germination of tile dormant buds and their development are also decided by the sites and the influence of infection of A mellea . In some cases, when the mixed bud of G. elara was injured, it could also form several tubers with the infection of A. mellea . 2. The infecting process of A. mellea on G. elata has been studied in detail. Some of our observations are not well fitted with Kusano's experimental results. We find that there are two modes of infection of A. mellea on G. clara: physiological infection and pathological infection. Under normal environmental conditions, A. mellea infects the tissues of cortical layer only, and may also lead to the enlargment and malformation of nuclei. On the other hand; the digestive cells possess both the functions of defense and infecting hyphae. The pathological infection often exists under unfavorable conditions. 3. The tuber of G. data is not the only organ to assimilate the A. mellea, the vegetative stem also has a function of assimilation. 4. The growihg tuber would not become an assimilating organ until winter was over.     

Study on Ultrastructure and Localization of Acid Phosphatase of Gastrodia elata Cortical Cells in Their Digestive Process of Armillaria mellea

In order to study the mechanism of the digestive process of Armillaria mellea in Castrodia data, electron microscopy and cytochemical method for determination of acid phosphatase activity was employed. The provacuoles were formed by means of expanded or convoluted ER under the stimulation of cortical cells and large cells of Gastrodia data by Armillaria mellea. A product of acid phosphatase (lead phosphate deposits) occured on the tonoplast. The papillae were produced in the cell wall of cortex in Gastrodia data when Armillaria mellea penetrated into its cortex. Our results showed that the enzyme was not released from cell of Armillaria mellea. A number of small vacuoles in the cortical cells disappeared. At the same time, lead phosphate deposits on the Armillaria mellea hyphae wall were observed and than Armillaria mellea hyphae wall was going to be digested, and the hyphae lost their structure. The activity of Armillaria mellea hyphae was not observed in the large cell of Gastrodia data. A great deal of small vacuoles and mitochondria were produced, at the same time the renewable nuclei and nuclolar vacuoles etc. appeared in the large cells of Gastrodia data under the stimulation of Armillaria mellea.     

蜜环菌成熟菌索的超微结构

用光学和电子显徽镜研究了生长在木材上的蜜环菌成熟菌索的超微结构。拟薄壁组织中有线粒体、内质网、核、液泡、质膜体、核糖体和典型的桶孔隔膜等。在菌索腔中疏松组织由薄壁菌丝、厚壁菌丝和非细胞衬质物质所组成,这些菌丝除了无隔孔帽的隔孔器和分散的核仁以外,见不到其它普通的细胞器,其最明显的特征是质壁分离和质膜断裂而卷曲,似乎是处在脱水状态。一旦它们进入新的寄主,一些普通细胞器就可出现。所以,可认为这些菌丝是处于休眠状态。对蜜环菌菌索的结构和功能的关系也进行了讨论。     

天麻大型细胞消化蜜环菌过程中溶酶体小泡的作用

蜜环菌(Armillaria mellea Fr.)菌丝由天麻(Gastrodia elata Bl.)皮层细胞经纹孔侵入大型细胞。初期大型细胞的原生质膜凹陷,同时细胞壁产生乳突状加厚阻止菌丝侵入。当菌丝侵入大型细胞以后,凹陷的质膜将菌丝紧密包围,大量由单位膜围成的小泡聚集在其周围。随后这些小泡的膜与质膜融合并将其内含物释放到菌丝周围的空间中,凹陷质膜逐渐膨大成为一个包围菌丝的消化泡。小泡和消化泡中均具酸性磷酸酶活性反应产物,证实其分别相当于植物溶酶体系统中的初级和次级溶酶体。菌丝在消化泡中被彻底消化。     

The Relation Between Growth of Gastrodia elata Protocorms and Fungi

Armillaria mellea penetrated protocorms from seed germination and vegetative multiplication corms of Gastrodia elata with rhizomorph. At beginning, they formed a hypha passing road and a hypha flow in the inner cells of cortex, and then, they both penetrated inside of large cells and penetrated outside of cortical cells. Gastrodia elata seeds depended on digesting Mycenct osmundicola etc gain nutrition to germinate at the stage of sexual reproduction, but its corms of vegetative multiplication must be penetrated by Armillaria mellea obtaining nutrition for normal growth at the stage of vegetative propagation.     

The Structure and Antifungal Functions of Vegetative Propagation Corm of Gastrodia elata

The growth of Gastrodia elata Bl. and Armillaria mellea (Vahl. ex Fr.) Quel. shares a special symbiotic relation. In general, A. mellea invades the G. elata , the epidermal cells, the cortical cells and the large cells of the growing vegetative propagation corm of G. elata . The empty cavity cells, the cork cells of the isolation in the vegetative propagation corms and the large cells of G. elata were the defensive structure, protecting the new G. elata from pathological invasion by A. mellea . In winter, G. elata enters the stage of hibernation.　The faulting layer derived from the cork cells of the isolation was the last defensive structure by which new G. elata could safely live through the winter.     

猪苓菌丝形成菌核结构的特点

对猪苓(Grifola umbellata(Pers.)Pilat)菌丝在人工条件下形成菌核及繁殖过程、人工菌核与野生菌核及培养基上未形成菌核的猪苓菌丝的显微结构进行了系统研究.研究证明:人工菌核的结构与野生菌核的结构相似,均具有菌髓和皮层结构.人工菌核中的菌丝与培养基表面未形成菌核的猪苓菌丝存在着显著的差异,人工菌核是由培养基上纯培养的菌丝分化为膨大菌丝再由此形成有高度组织分化的猪苓菌核.     

猪苓菌丝形成菌核结构的特点(英文)

对猪苓(Grifolaumbellata(Pers.)Pilat)菌丝在人工条件下形成菌核及繁殖过程、人工菌核与野生菌核及培养基上未形成菌核的猪苓菌丝的显微结构进行了系统研究。研究证明人工菌核的结构与野生菌核的结构相似,均具有菌髓和皮层结构。人工菌核中的菌丝与培养基表面未形成菌核的猪苓菌丝存在着显著的差异,人工菌核是由培养基上纯培养的菌丝分化为膨大菌丝再由此形成有高度组织分化的猪苓菌核。     

天麻球茎不同部位的细胞对蜜环菌侵染的反应

用电镜技术比较研究了天麻球茎皮层细胞和中柱细胞对蜜环菌侵染的反应,皮层细胞在蜜环菌菌丝的刺渺下能产生囊状结构,然后这些小囊将入侵的菌丝包围和消化,而中柱细胞则不能,上述两种细胞在功能上的主要差别就在于此。     

Morphological characteristics of sclerotia formed from hyphae of <Emphasis Type="Italic">Grifola umbellata</Emphasis> under artificial conditions

The morphological characteristics of sclerotia were induced in cultures of the fungus Grifola umbellata by introducing an unidentified companion fungus were studied by light microscope, scanning and transmission electron microscope (SEM and TEM). Light microscope and SEM investigations of developing sclerotia revealed that aerial mycelial hyphae diminished with age, and mature sclerotia had two tissue layers, the rind and medulla. The medulla was comprised of thin and thick-walled hyphae of varying diameter. The thick-walled cells always formed below the hyphal tips. Retraction of the cytoplasm was accompanied by the thickening of cell wall. There were crystalline initials in the newly formed sclerotium. Crystalline initials were always formed in the tip of medullary hyphae and were not of regular shape. A series of changes occurred in the cells in which the crystalline initials would be formed, such as enlargement of size, formation of one or several large vacuoles. Crystalline initials developed via amorphous materials in the cytoplasm deposited in the vacuoles. At last crystalline initials was released by degradation of the cell wall.     

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。