天麻吸收蜜环菌营养机制的细胞学研究

天麻(Gastrodia elata BI.)地下块茎皮层内具三种染菌细胞:通道细胞、寄主细胞和消化细胞。超微结构的研究表明,通道细胞被真菌所破坏,寄主细胞与真菌保持共生关系,而消化细胞能反寄生于真菌并从真菌摄取营养。消化细胞首先释放溶酶体小泡消化真菌,然后通过内吞管和内吞泡吸收菌丝细胞质降解后渗漏的可溶性有机大分子物质,后期通过消化泡进一步吞噬和消化不溶性菌丝细胞壁物质。     

蜜环菌和天麻共生营养关系的放射性自显影研究

本文利用＾３Ｈ－葡萄糖以打孔浇灌法标记天麻。用标记的天麻伴栽蜜环菌,追踪标记化合物是否存在蜜环菌中。显微放射自显影的结果表明：天麻自身能吸收标记物,同化的标记物其生距离运输靠天麻的纵向维管组织进行。天麻表皮外的蜜环菌菌索的光镜、电镜自显影显示,蜜环菌能从天麻中获取营养,蜜环菌和天麻之间存在营养物质的相互交流,菌麻之间存在着特殊的共生关系。     

蜜环菌和天麻共生营养关系的放射性自显影研究

本文利用3H-葡萄糖以打孔浇灌法标记天麻。用标记的天麻伴栽蜜环菌,追踪标记化合物是否存在蜜环菌中。显微放射自显影的结果表明：天麻自身能吸收标记物,同化的标记物其长距离运输靠天麻的纵向维管组织进行。天麻表皮外的蜜环菌菌索的光镜、电镜自显影显示,蜜环菌能从天麻中获取营养,蜜环菌和天麻之间存在营养物质的相互交流,菌麻之间存在着特殊的共生关系。     

供给天麻种子萌发营养的真菌——紫萁小菇

从天麻种子发芽的原球茎中,共分离到12种可供给天麻种子萌发营养的真菌,其中最优良的01号菌株,诱导培养出子实体,经鉴定为紫萁小菇(Mycena osmundicola Lange),属国内新纪录。用01号菌株及紫萁小菇培养的染菌树叶,伴播天麻种子都可发芽,经酯酶同工酶凝胶电泳分析,二者酶带条数、迁移率(RF值)基本相同,证明鉴定结果可靠(标本保存在医科院药植所真菌研究室)。     

蜜环菌菌种分离新法——天麻组织分离法

报道了一蜜环菌菌种分离方法——天麻组织分离法,并对此法与常用分离法——菌索分离法进行比较试验。结果发现,天麻组织分离法分离成功率高（78％）,远高于常用的菌索分离法（16％）,且前者操作简便、难度低,所得菌种生活活力、生长形态均优于后法。     

不同蜜环菌菌株对乌天麻产量的影响

用不同的蜜环菌(Amillaria mellea)菌株与长白山乌天麻(Gastrodia elata)进行伴栽试验,以探索不同蜜环菌菌株对乌天麻产量的影响。结果表明:不同蜜环菌菌株对乌天麻产量的影响不同,其中菌株Ar-4能显著提高天麻产量,仿野生栽培和室外箱栽试验均表现出良好的伴栽效果。     

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。     

乌天麻Gastrodianin基因的鉴定及其在天麻发育进程中的表达

基于相似性克隆策略和RT-PCR,从兰科植物乌天麻(Gastrodia elataf.glauca)球茎克隆得到两个新的天麻抗真菌蛋白基因,命名为gastrodianin-4A(ga4A)和gastrodianin-4B(ga4B),并首次用Northern杂交研究了该基因在植物不同部位的转录表达。结果表明,天麻个体的不同部位只转录表达同一种Gastrodianin基因,纯化自球茎的Gastrodianin蛋白质的肽质量指纹谱与推导的Ga4A和Ga4B成熟蛋白的氨基酸序列相吻合;Northern杂交证明天麻的地上器官Gastrodianin基因转录表达量远远高于地下球茎,而次生球茎皮层组织的表达量比中柱和整个营繁茎的都高一些。天麻球茎Gastrodianin的外周表达模式可能是天麻在地下抵御蜜环菌(Armillaria mellea Karst)入侵球茎皮层内部的防卫机制之一,但该基因在天麻地上部分的高丰度表达暗示Gastrodianin可能蕴藏其它生理功能。     

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。     

天麻生长过程中酸性磷酸酶的研究

以蜜环菌(Armillaria mellea)伴生的天麻(Gastrodia elata)块茎为材料,研究了天麻生长发育过程中酸性磷酸酶(ACP)活性及其同工酶的变化。结果表明,在天麻生长90d时,其酶谱中有3条谱带,且均显色较弱;生长至120d时,其酶谱与生长90d的酶谱相同,但谱带显色加深;而在生长150d时,酶谱中缺少了一条Rf 0.852的谱带。在90~150d的天麻生长发育过程中,酸性磷酸酶(ACP)活性呈现先升高后下降的趋势。讨论了酸性磷酸酶(ACP)同工酶变化的生理学意义。     

L-异亮氨酸产生菌的选育

为检测不同产地天麻中氨基酸的含量差异,采用日立L-8800氨基酸自动分析仪对7个分别产自四川荥经县、剑阁县和云南小草坝的野生及引种天麻品种的氨基酸含量进行了测定.结果显示,7种天麻的氨基酸种类丰富且含量均较高,其中以四川荥经县野生样品中氨基酸含量最高,四川荥经县引种云南小草坝样品中氨基酸含量最低.结果表明,不同产地的天麻中氨基酸的含量存在差异.     

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.     

A Cytological Study on the Nutrient Uptake Mechanism of a Saprophytic Orchid Gastrodia elata

There are three type cells infected by the mycorrhizal fungus, ArmiUaria mellea (Vahl ex Fr. ) Karst in the tube cortex of Gastrodia elata BI., namely the passage cells, host cells and digesting cells. Ultrastructural study demonstrated that the passage cells were distroyed by the hyphae, the host cells kept symbiotic with the hyphae, but the digesting cells could become inversely parasitic on the hyphae from which nutrient were being uptaken. The detail process of the digesting cells obtaining nutrient'from the fungus is described as follows: Firstly the digesting cells began to attack the invading hyphae by releasing numerious electron-transparent vesicles of lysosomal property, secondly they took up the soluble organic material leaked out from the digested hyphae by forming many electron-dense endocytic tubes and vesicles, and finally they endocytosed and hydrolysed the insoluble hyphal walls by forming large digesting vacuole in which a piece of hyphal wall was completely enveloped.     

黔产天麻的化学成分

从黔产天麻（Ｇａｓｔｒｏｄｉａ ｅｌａｔａ Ｂｌ．）中分离得到１个新的含氮化合物,命名为天麻羟胺（ｇａｓｔｒｏｄａｍｉｎｅ,Ⅰ）,经波谱分析,确定其结构为双－（对羟苄基）羟胺。除β－谷甾醇和胡萝卜甙外,同时还分离鉴定了３个已知化合的：Ｌ－焦谷氨酸（Ⅱ）、柠檬酸单甲酯（Ⅲ）、柠檬酸双甲酯（Ⅳ）,其中化合物Ⅱ和Ⅳ为首次报道从天麻中得到的。     

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.     

天麻的营养繁殖茎及其抑菌功能

天麻9Gastrodia elata Bl.）与蜜环菌（Armillaria mellea (Vahl.ex Fr.)Quel.)营共生生活,在正常情况下蜜环菌只侵染种麻及分化生长出的营养繁殖茎的表皮、皮层及大型细胞层。种麻的大型细胞层及营养繁殖茎隔离区的空腔细胞层和木栓细胞层,都是种麻与新生麻的防御结构,它们保护新生麻不遭蜜环菌病理侵染而正常生长。冬季,天麻进入冬期,隔离区的木栓细胞层形成断裂层,此层细胞是新生麻能够安全越冬的最后一道防御结构。