以豆粕为氮源生产虫草菌丝体的研究

采用豆粕粉取代蚕蛹粉作氮源进行了虫草菌丝体的发酵试验,生产出无异臭味的虫草菌丝体,扩大了它的应用范围;对虫草菌丝体的抽提液进行超滤处理,测定了虫草多糖的含量和分子量。     

菌核侧耳液体培养条件优化研究

运用液体深层培养法,采用不同pH值、不同碳源和氮源对菌核侧耳菌丝体生长的影响进行了探讨。结果表明:菌核侧耳菌丝体生长的培养基适宜pH值为5.0～6.5,葡萄糖、蔗糖、乳糖和纤维素是菌核侧耳菌丝体生长的适用碳源;黄豆、麦麸、酵母膏和蛋白胨等为适宜氮源;其中以黄豆浆作为氮源,蔗糖作为碳源,初始pH5.0为最佳条件组合。     

发酵法生产猪苓菌丝体及猪苓多糖的研究

采用经选育的猪苓PU－99菌作为生产菌株,研究了其培养基组成,优化了深层发酵条件。在1吨罐中生产猪苓菌丝体：发酵36h,其菌丝体干重达2．3％,粗多糖含量在31．0％。对得到的猪苓菌丝体进行了有效成分的提取,并测定了猪苓多糖的分子量,分子量为29054,峰面积72．02％。     

平菇菌丝体与子实体营养成分的分析比较

采用现代化学分析手段对平菇菌丝体和子实体的营养成分进行分析比较。结果表明,用PDA固体培养的菌丝体的灰分、可溶物、总糖和粗蛋白的含量均明显低于子实体,而粗脂肪和粗纤维的含量则显著高于子实体;菌丝体的Fe、Ca、Zn的含量显著高于子实体,而Mg和K的含量明显低于子实体,Na的含量两者比较接近;菌丝体和子实体的氨基酸组成相似,都含有17种氨基酸,但子实体的各氨基酸的含量均高于菌丝体,从各项氨基酸含量的比值来看,菌丝体则更优于子实体。两者都不失是极好的营养食品源。     

桦褐孔菌三萜类物质的提取与含量测定

以桦褐孔菌发酵菌丝体为材料,通过对溶剂乙醇(95%)、甲醇、乙酸乙酯、丙酮、异丙醇、正己烷和氯仿的提取效果比较表明,提取三萜类化合物的最佳溶剂为异丙醇,提取时间为24h,每个样品所需溶剂量(6mL)和菌丝体样品量(100mg)较少,并可同时对大量样品进行有效提取。以白桦脂醇为标准品,对香草醛-冰醋酸-高氯酸分光光度法进行评价,证明该方法简单快速、准确度高、试验误差小、重复性好。利用此方法对桦褐孔菌的野生菌核和发酵菌丝体内三萜类化合物含量进行测定,结果表明野生菌核(59.86mg/g)和发酵菌丝体(94.92mg/g)中都含有很高的三萜类化合物,且发酵菌丝体中三萜类化合物含量高于野生菌核。因此在桦褐孔菌产品开发中,发酵菌丝体可代替野生菌核进行大工业化生产。     

桦褐孔菌三萜类物质的提取与含量测定

以桦褐孔菌发酵菌丝体为材料,通过对溶剂乙醇（95%）、甲醇、乙酸乙酯、丙酮、异丙醇、正己烷和氯仿的提取效果比较表明,提取三萜类化合物的最佳溶剂为异丙醇,提取时间为24h,每个样品所需溶剂量（6mL）和菌丝体样品量（100mg）较少,并可同时对大量样品进行有效提取。以白桦脂醇为标准品,对香草醛-冰醋酸-高氯酸分光光度法进行评价,证明该方法简单快速、准确度高、试验误差小、重复性好。利用此方法对桦褐孔菌的野生菌核和发酵菌丝体内三萜类化合物含量进行测定,结果表明野生菌核（59.86mg/g）和发酵菌丝体（94.92mg/g）中都含有很高的三萜类化合物,且发酵菌丝体中三萜类化合物含量高于野生菌核。因此在桦褐孔菌产品开发中,发酵菌丝体可代替野生菌核进行大工业化生产。     

鲍姆纤孔菌（桑黄）不同方式发酵的菌丝体生物活性比较

利用液体发酵、木屑固体发酵和米饭固体发酵3种方式培养鲍姆纤孔菌（桑黄）菌丝体,对菌丝体醇提物的体外抗氧化、抗肿瘤和抗衰老生物活性进行了研究。结果表明,木屑固体发酵、液体发酵和米饭固体发酵的菌丝体醇提物清除H2O2自由基的IC50值分别为78.28±0.32、27.73±0.57和7.84±0.37;米饭培养的桑黄菌丝体醇提物在低浓度500μg/mL下对超氧阴离子自由基清除作用到达80%,在相同的浓度下对DPPH自由基清除率也明显高于其他两种方法,表现出较高的抗氧化活性。木屑以及米饭培养方法得到的菌丝体对PC12神经细胞损伤修复均有较好的效果,液体培养的桑黄菌丝体表现的修复作用较低;液体发酵培养的菌丝体醇提物浓度在100μg/mL时,对肿瘤细胞HepG2的抑制率达70%,高于其他两种培养方法的抑制作用。     

粗柄鸡菌菌丝体培养和无性繁殖过程研究初报

鸡菌属菌物是一类具有商业前景的食物蕈菌。对Ｔｅｒｍｉｔｏｍｙｃｅｓａｌｂｕｍｉｎｏｓｕｓ（Ｂｅｒｋ）Ｈｅｉｍ进行了广泛研究,其它种类的研究仅见于Ｔ．ｆｕｌｇｉｎｏｓｕｓ（Ｂｅｒｋ．）Ｈｅｉｍ。笔者首次对Ｔ．ｒｏｂｕｓｔｕｓ（Ｂｅｅｌｉ）Ｈｅｉｍ粗柄鸡菌进行了纯培养研究,其菌丝体生长所需的碳源以麦芽糖为佳,葡萄糖、蔗糖次之;氮源以蛋白胨、酵母粉较好;天然材料的热水提取物,如松针、蚁巢圃、马铃薯、平菇出菇废料、蚕豆的混合物能促进菌丝生长;培养基在ｐＨ４．５左右最佳,可用０．０５％的柠檬酸调节。菌丝体纯培养很容易形成大量分生孢子和小白球菌,表明小白球菌是粗柄鸡菌菌丝体无性繁殖过程的一个阶段。     

灰树花发酵工艺及培养基研究

详细、系统地研究了碳源、氮源、无机盐、维生素以及种龄、接种量、摇床转速、摇瓶装液量和培养基 pH值等因素对灰树花液体深层发酵菌丝体产量的影响 ,结果表明 :玉米粉、葡萄糖为最佳碳源 ,豆饼粉为最佳氮源 ,KH2 PO4 ,MgSO4 以及少量VB1的添加均使菌丝体产量明显增加     

草菇培养物中粗三萜和黄酮含量及抗氧化抗肿瘤活性研究

采用液体摇瓶法培养草菇菌丝体,并用不同有机溶剂对培养液及菌丝体中的代谢成分分别进行分离提取,获得了不同来源的次生代谢提取物。对各提取物的成分分析表明,石油醚相、乙酸乙酯相和乙醇相提取物中均含有粗三萜和黄酮类物质,但菌丝体提取物中粗三萜和黄酮含量明显高于培养液提取物中的含量。石油醚抽提菌丝体获得的提取物中粗三萜含量最高,达17%,而乙酸乙酯抽提菌丝体获得的提取物中黄酮含量最高,达9.31%。抗氧化活性检测结果显示,石油醚相、乙酸乙酯相、乙醇相中的代谢提取物均有较高的抗氧化活性,但乙酸乙酯相提取物的抗氧化活性明显高于石油醚相提取物,具最高抗氧化活性的提取物分别来自乙酸乙酯、乙醇对菌丝体的抽提物。MTT法检测各提取物对胃癌细胞增殖的抑制作用,结果显示各组分均有较高的抗肿瘤活性,且抗肿瘤活性与提取物浓度存在明显的量效关系。     

柿树炭疽菌侵染寄主的细胞学研究*

超微结构研究表明,柿树炭疽菌(Colletotrichum gloeosporioides)侵染后在寄主细胞中形成初生菌丝和次生菌丝,寄主细胞膜外沉积了一层厚的电子不透明物质,初生菌丝与具有沉积物的寄主原生质膜之间有一层界面基质(interfacial matrix)。当初生菌丝扩张并侵染相邻细胞时, 围绕着初生菌丝层的界面基质消失,具有沉积物的原生质膜被逐步降解。初生菌丝在穿透寄主细胞壁过程中形成一个漏斗状的菌丝锥,然后穿透寄主细胞壁并迅速膨大, 然后形成厚壁的初生菌丝。初生菌丝在寄主细胞壁中收缩狭窄处产生一个隔膜,隔膜两边菌丝中细胞质的电子密度明显不同,菌丝锥中有浓密的电子密度。死体营养的次生菌丝在死的细胞中繁殖和扩展,并产生分枝。次生菌丝可直接穿透较薄的寄主细胞壁,无缢缩或任何变形现象,菌丝顶端部分未见隔膜产生;在穿透较厚的细胞壁时,靠近顶端处产生隔膜,顶端细胞膨大,使寄主细胞壁撕裂。接种90h后分生孢子盘在枝条表面形成。柿树炭疽菌其侵染过程有两个阶段,即初生菌丝的活体营养阶段和次生菌丝的死体营养阶段。     

柿树炭疽菌侵染不同柿树种、品种和部位的细胞学特征

用柿树炭疽病菌Colletotrichumgloeosporioides的分生孢子制备孢子悬浮液,接种无核柿、野柿、冬柿和浙江柿的新梢、叶柄和叶片,并观察致病性、附着胞形成和侵染特性。柿树炭疽菌可以侵染无核柿枝条和叶柄以及野柿枝条,但不侵染无核柿叶片、野柿叶柄和叶片,也不侵染冬柿和浙江柿枝条、叶柄和叶片。室内接种试验与田间自然发病结果一致。柿树炭疽菌在不同柿树表面均能形成附着胞,附着胞产生在寄主表皮背斜细胞壁间结合处(JACWs)或近结合处的百分率达81%~93%。接种12h后,不同柿树表面都有附着胞形成;36h后,无核柿枝条、叶柄中有侵染菌丝存在;48h后,无核柿枝条、叶柄中观察到膨大初生菌丝和较细次生菌丝,初生菌丝可扩展到相邻细胞中,而野柿枝条中仅观察到侵染菌丝;60h后,野柿枝条中也观察到膨大的初生菌丝和较细的次生菌丝,但初生菌丝仅局限在最初侵染的细胞中,无核柿枝条和叶柄以及野柿枝条中都有分枝的次生菌丝在细胞内、细胞间或相邻的细胞中扩展;直到接种90h后,在冬柿和浙江柿上都未观察到侵染菌丝的形成。结果表明,柿树炭疽菌在不同柿树种和品种上侵染菌丝的形成和扩展方式可能是其寄主专化性(或致病性)差异的重要机制之一。     

菌核侧耳菌丝染色方法的研究

在对菌核侧耳单双核菌丝进行染色方法的研究中, 通过采用不同的菌丝培养方法, 比较2种固定液及3种染液的染色效果, 优化出菌丝染色方法, 进而得出一套稳定可靠的鉴定菌丝的细胞核染色技术。     

Rhizomorph anatomy of <Emphasis Type="Italic">Ossicaulis lignatilis</Emphasis> (Tricholomatales), with special attention to its haustoria-like intrahyphal hyphae

The stout, basidiomal rhizomorphs of the agaricoid Hymenomycete Ossicaulis lignatilis are tightly woven from thin, generative hyphae, broad tubular hyphae, and broad thromboplerous hyphae that contain numerous thin, haustorialike, intrahyphal hyphae in their deuteroplasm. The plectology and cytology of the rhizomorphs and a possible role of the thromboplerous hyphae are described and discussed.     

The solid phase attachment of fungal hyphae in an ELISA to screen for antifungal antibodies

A method to immobilize fungal hyphae onto the wells of 96-well microplates for use in an in-direct ELISA to screen for antifungal antibodies in sera and cell culture supernatants is described. The hyphae from three genera (Penicillium, Eurotium andFusarium) were successfully attached by overnight drying onto wells precoated with poly-L-lysine and glutaraldehyde. Microscopy revealed that the hyphae remained attached to the wells throughout the ELISA and antiserum titrations showed that the attached hyphae were uniformly coated and remained reactive. Background absorbances were low and the plates could be stored at –20 °C without loss of reactivity.     

小麦条锈菌胞间菌丝的超微结构和细胞化学研究

本文应用电镜技术和细胞化学方法,对小麦条锈菌寄主胞间菌丝的超微结构进行了研究。观察发现:小麦条锈菌胞间菌丝有两种类型,即具隔膜菌丝和无隔膜菌丝。在胞间菌丝中,多核现象极为普遍。常规染色和细胞化学染色结果表明:胞间菌丝的细胞壁由四层组成,隔膜由三层构成,细胞壁的内层与隔膜的外层相连,细胞壁和隔膜中含有蛋白质和多糖物质。隔膜的发育可分三个阶段,即隔膜突的形成,隔膜壁的延伸和隔膜孔结构的形成。本研究中还观察到胞间菌丝间的融合现象。本文的研究结果表明:小麦条锈菌胞间菌丝的一些特征显然不同于其它锈菌。     

Compared Anatomy of Young Leaves of Prunus persica (L.) Batsch with Different Degrees of Susceptibility to Taphrina deformans (Berk.) Tul

Anatomical observations of leaves infected by Taphrina deformans were studied in tolerant peach trees (TPT) and in very susceptible (VSPT) ones. Leaves from the first sampling (2nd April) showed hyphae penetrating through the stomata or into the cuticle of the host tissue; anatomical structures of leaf sections were similar for both TPT and VSPT. The ultrastructure of the leaves of TPT showed seemingly normal mesophyll cells. In contrast, mesophyll cells of the VSPT showed important signs of degradation. Cells were organelle‐free and the middle lamella was expanded and invaded by hyphae of T. deformans. In some samples, the leaves of TPT showed deformed epidermal cells, loss of some spongy cells and increase of the intercellular spaces and division of the palisade cells. The pathogen proliferation in the leaves of the VSPT was considerably superior. In this case, stimulation of cell division occurred in the abaxial epidermis. Cells showed periclinal and oblique divisions, with an increased number of plasmodesmata; palisade or spongy cells were not differentiable. Leaves from TPT collected on 26th April showed hyphae with a non‐cylindrical section and with a squashed aspect. The hyphae were very evident in the intercellular spaces, showing abundant endoplasmic reticulum of rough type (RER) in the cytoplasm. On the other hand, epidermis of the leaves of the VSPT had numerous hyphae under the cuticle, which were growing in a thick pectin matrix. Leaves from TPT and VSPT collected on 6th May showed relevant differences. The leaves of TPT had a palisade mesophyll with fewer cells but with active chloroplasts. In contrast, the leaves from VSPT showed empty mesophyll cells, the cytoplasm was collapsed and the adaxial epidermis was covered with the fungus fructification. The observed anatomical and ultrastructural differences of leaves from TPT and VSPT confirm a different behaviour in plant‐host reaction at early stages of infection.     

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。     

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.     

松口蘑人工菌塘诱导形成的初步研究

本研究通过土壤筛选找到适合松口蘑菌丝体生长的土壤,并在其土壤中添加适当浓度的橄榄油(0.5%-2.0%),有效地促进了菌丝体在土壤中的生长,菌丝生长状况酷似野外的松口蘑菌塘,且随着添加浓度的增加,菌丝体生物量呈明显的正相关。当土壤中橄榄油浓度为0.5%和1.0%时,松口蘑共生植物赤松苗的生长不但没有受到抑制,而且还能与菌丝体在这种“菌塘”中形成菌根。本研究成功地建立了中国松口蘑人工菌塘诱导体系,同时也为其人工子实体的诱导研究提供了必要的前提条件。