2种真菌多糖荧光标记及其取代度和稳定性

通过对小刺猴头发酵浸膏多糖和黑木耳发酵浸膏多糖进行荧光标记,得到2种真菌多糖的荧光标记产物,即小刺猴头菌多糖-FITC和黑木耳多糖-FITC。通过紫外分光光度计、酶标仪和荧光分光光度计对标记后的真菌多糖进行扫描,并进行运算,得到该2种真菌多糖的荧光物质的取代度,分别为0.79%和0.71%。同时对标记后多糖的体外稳定性进行研究,结果表明,小刺猴头发酵浸膏多糖标记物和黑木耳发酵浸膏多糖标记物在PBS溶液、血浆及尿液中稳定性良好。     

小刺猴头菌多糖的荧光标记及其稳定性

通过体外和小鼠体内试验,验证荧光标记的小刺猴头菌多糖的稳定性和生物体内代谢途径。结果表明:标记后的小刺猴头菌多糖体外稳定性在24 h内良好,小鼠体内代谢途径主要通过消化系统代谢,并在8 h内基本代谢完毕,这一结果为荧光标记真菌多糖的药理活性提供了依据。     

小刺猴头菌发酵液小分子指纹图谱与多糖含量相关性研究

试验研究了小刺猴头菌发酵液小分子指纹图谱与多糖含量的相关性。在小刺猴头菌摇床培养发酵液整个发酵周期内取得的样品经石油醚、二氯甲烷和乙酸乙酯依次萃取,HPLC检测分析。选择二氯甲烷层的3种小分子物质含量为主要研究对象,通过SAS软件和SIMCA-P 11.5软件对其与发酵液多糖进行相关性分析。结果表明:发酵液二氯甲烷层的3,4,5号峰小分子与发酵液多糖在含量变化上呈极显著相关(P<0.01)。     

小刺猴头菌液体体深层培养过程中多糖积累及其特征的研究

本文探讨了所升环流反应器培养小刺猴头菌过程中的菌体生物量、胞内外真菌多糖的积累规律,与同一菌株复元的子实体多糖的含量、组成分别进行分析对比,得到液体深层培养菌丝体多糖的含量为０．０９１８ｇ／ｇ,而子实体仅为０．０３７８ｇ／ｇ,发酵液为多糖含量达０．２８５ｇ／ｌ,它们的单糖构成比例均相同,即葡萄糖：甘露糖：半乳糖＝２０．２：５．２：１,但液体深层培养的菌丝体多糖与人工栽培的子实体多糖的电荷性能不同。     

猴头菌子实体发育过程中多糖结构和活性的变化

猴头菌(Hericium erinaceus)多糖是猴头菌发挥药理作用的主要活性成分,猴头菌生长过程中多糖的合成代谢及其结构的变化目前还鲜有研究报道。该试验研究了猴头菌生长发育过程中多糖的产生及其结构和活性的变化。对猴头菌0605菌株生长发育6个时期的子实体进行了多糖和β-葡聚糖含量的测定,并对提取的粗多糖进行了体外免疫活性的研究。结果表明:在猴头菌的整个生长发育过程中,子实体多糖的含量呈现上升的趋势,成熟期含量最高,达1.54%,分裂期含量最低仅0.89%;β-葡聚糖含量整体上呈上升的趋势,中菌刺期含量最高33.58%,成熟期最高,达到50.67%;体外免疫活性试验表明,现蕾期、分裂期、中菌刺期和成熟期的多糖均能很好地刺激巨噬细胞RAW264.7产生NO,活性均优于无菌刺期和小菌刺期的多糖。分别采用体积分数为30%、50%和70%的乙醇对子实体发育后3个时期的热水浸提物进行分级醇沉,对分级分离多糖进行理化性质研究。结果表明:小菌刺期30%醇沉的粗多糖得率高于中菌刺期和成熟期,其大分子量多糖的比例也较高;3个时期50%和70%醇沉多糖分子量相差不大;分级分离多糖的单糖种类基本相同,都含有岩藻糖、半乳糖、葡萄糖和甘露糖,但摩尔比有差异;3个时期分级分离多糖均能刺激巨噬细胞分泌NO,30%醇沉多糖比50%和70%醇沉多糖的活性好;成熟期多糖的活性优于其他生长阶段。对小菌刺期、中菌刺期和成熟期50%醇沉多糖(H5FP4B、H5FP5B、H5FP6B)进行Sephacryl S300凝胶层析纯化,共得到9个组分:H5FP4B-1、H5FP5B-1、H5FP6B-1、H5FP4B-2、H5FP5B-2、H5FP6B-2、H5FP4B-3、H5FP5B-3、H5FP6B-3。对这9个组分的分子量分布进行研究,发现每个时期的主峰(H5FP4B-3、H5FP5B-3和H5FP6B-3)分子量基本相同,分子量分布差异较大的是3个时期的第1组分,在体外免疫活性上也表现出较大差异。主峰H5FP4B-3的分子量为1.59万,由岩藻糖、半乳糖和葡萄糖组成,摩尔比为5.2∶23.9∶1。经红外光谱、甲基化和核磁共振综合分析,解析出H5FP4B-3均一多糖的结构单元是以α-D-1,6连接的半乳糖和α-D-1,2,6连接的半乳糖构成主链,侧链为α构型的端基岩藻糖。研究结果表明:猴头菌不同发育阶段产生的多糖结构和活性有一定的差异,生长初期,大分子量的多糖较多,后期分子量分布差异变小并趋稳定。多糖主成分变化不大,较稳定;到成熟期,多糖的活性更高。     

小刺猴头菌液体深层培养过程中多糖积累及其特征的研究

本文探讨了气升环流反应器培养小刺猴头菌过程中菌体生物量、胞内外真菌多糖的积累规律,与同一菌株复元的子实体多糖的含量、组成分别进行分析对比,得到液体深层培养菌丝体多糖的含量为0．0918g／g（干）,而子实体仅为外0．378g／g（干）,发酵液中多糖含量达0．285g／l,它们的单糖构成比例均相同,即葡萄糖：甘露糖：半乳糖＝20．2：5．2：1,但液体深层培养的菌丝体多糖与人工栽培的子实体多糖的电荷性能不同。     

猴头菌不同发育阶段产生的多糖结构特征及免疫活性

采收猴头菌Hericium erinaceus 7个不同发育阶段的子实体,经热水浸提后分别采用20%、50%、70%的乙醇终浓度进行分级沉淀,获得21个多糖组分,对它们的结构特征及体外免疫活性进行了研究。结果表明,在猴头菌发育过程中,多糖总得率呈现先增大后减小的趋势,在第5阶段达到最大值0.92%。20%醇沉的多糖含量也呈先增大后减小的趋势,在第5阶段达到最大值42.87%,其大分子量多糖（1 000-5 000kDa）所占比例最高。50%、70%醇沉多糖均为小分子量多糖,约为10-40kDa。获得的猴头菌多糖组分中,单糖组成多以岩藻糖、半乳糖、葡萄糖和甘露糖为主（相对比例存在一定差异）;另外,第1阶段20%、70%醇沉多糖还含有少量的核糖,第7阶段20%醇沉多糖含有一定量的鼠李糖;7个时期50%醇沉多糖均含有一定量的葡萄糖醛酸。所得多糖样品均具有刺激巨噬细胞释放NO的活性,其中20%醇沉多糖的活性优于50% 和70% 醇沉多糖,在50μg/mL时就表现出显著的体外免疫活性。此外,在第5阶段即中菌刺期产生的多糖活性最优,说明在此阶段采摘可以获得最佳的猴头菌多糖原材料。本研究为猴头菌生长发育过程中活性多糖的动态形成研究提供了一定的理论基础。     

胎儿发育过程中食管上皮糖复合物的改变

采用十种生物素化凝集素 U EA- I、 DBA、 PSA、 BSL、 PNA、 L CA、 RCA- I、SBA、 Con A及 WGA分别对 8W、1 4 W、 2 0 W　 2 8W及 32 W的人胎儿食管上皮进行研究 ,以确定胎儿食管上皮在发育过程中凝集素的结合位点以及结合方式随年龄的变化。结果提示 ,BSL、 RCA- I、 WGA的染色强度及特性不受年龄影响 ,而与之相对应 ,U EA - 1在除 2 0 W以外的食管上皮显色 ;含α- D- Glc NAc的糖复合物 (DBA受体 )只在 2 0 W表达 ;含α- D- man/α- D- Glc的糖复合物 (PSA受体 )则只出现于 32 W;Con A受体虽在胎儿食管上皮各年龄组都表现阳性反应 ,但在 2 8W 时受体在上皮各层细胞中的分布方式及染色强度出现了有意义的变化 ,即腔面细胞染色阴性 ,中层细胞染色强阳性 ,基底细胞弱阳性 ,而在其他时间腔面反应强阳性 ,中层中等阳性 ,基底弱阳性 ;PNA、SBA及 L CA在各年龄组均呈阴性反应。这些结果表明 ,在胎儿食管上皮发育过程中存在着多个凝集素结合位点 ,含 α- L- Fucose、 α- D- Gal NAc、α- D- Man/ α- D- Glc残基的糖复合物在分布方式和 /或量上存在着发育相关性改变 ,即糖复合物的改变具有阶段性。总之 ,本实验资料提示 ,胎儿食管上皮在发育过程中表现出不同的结合位点 ,RCA- 、 BSL、 WGA受体似与年龄变     

小刺猴头菌发酵浸膏寡糖对荷瘤小鼠免疫功能的影响

通过建立S180荷瘤小鼠试验动物模型,研究小刺猴头菌发酵浸膏寡糖对S180荷瘤小鼠免疫功能的影响。选择灌胃的方法将不同组别小刺猴头菌发酵浸膏寡糖喂服到荷瘤小鼠体内,观察肿瘤生长情况,对其进行免疫功能的测定。结果表明:小刺猴头菌发酵浸膏寡糖可增加荷瘤小鼠免疫器官脾脏和胸腺质量,提高荷瘤小鼠血清的IL-2和TNF-α含量,抑制肿瘤生长,提高机体免疫功能。     

真菌多糖抗氧化活性的研究进展

真菌多糖的抗氧化作用目前正成为国内外众多学科领域研究的热点之一,对真菌子实体多糖、菌丝体多糖和胞外多糖的抗氧化活性研究现状进行了综述,并对其研究前景进行了展望,旨在为真菌多糖的进一步研究和利用提供参考.     

10种针阔叶树上常见的木材腐朽菌的培养特性*

描述了10种针阔叶树上常见的多孔菌的培养特性。它们是环纹革盖菌、蔷红拟层孔菌、盘拟层孔菌、斑褐孔菌、全缘孔菌、小刺猴头、黄白卧孔菌、烟色刺耳、变红栓菌和红斑干酪菌。     

Wide distribution of cellobiose-oxidizing enzymes in wood-rot fungus indicates a physiological importance in lignocellulosics degradation

When grown on lignocellulosic materials, cellobiose-oxidizing enzyme activities were detected in Coriolus versicolor, Flammalina velutipes, Ganoderma gibbasum, Hericium erinaceus, Neurospora crassa and Schizophyllum commune. No cellobiose-oxidizing enzyme activities were detected in 14 other cellulolytic fungi which are recognized as having no or very low lignin-degrading ability.     

小刺猴头菌丝体多糖的结构研究

对小刺猴头过滤掉发酵液的发酵菌丝体,水提和碱提后获得的均一组分多糖HMP-w1.1和HMP-a1.1进行结构性质的研究。结果表明:HMP-w1.1是分子量为36.3 kD的α型吡喃糖,单糖组成为甘露糖(Man),葡萄糖(Glc),半乳糖(Gal),岩藻糖(Fuc);HMP-a1.1是分子量为42.8 kD的β型吡喃糖,单糖组成为甘露糖(Man),半乳糖醛酸(GalUA),葡萄糖(Glc),半乳糖(Gal),岩藻糖(Fuc)。综合高碘酸氧化和Smith降解的试验结果,推断HMP-w1.1的糖苷键构型可能为1→、1→4、1→4,6、1→6、1→2、1→2,6;HMP-a1.1的糖苷键构型可能为1→6、1→2、1→2,6。     

Revision of Taeniatherum (Poaceae)

Three species are often accepted in Taeniatherum Nevski. Analysis of morphological characters show that indistinct limits occur between them. All taxa are found to be diploid (2n=14), with the same karyotype, but artificial hybridization shows genetical barrier to exist. For these reasons subspecific level is chosen for the taxa, viz. T. caput-medusae (L.) Nevski ssp. caput-medusae, T. caput-medusae ssp. crinitum (Schreb.) Melderis and T. caput-medusae ssp. asperum (Simk.) Melderis. Map showing known distribution of the subspecies is presented.     

四川大型真菌资源调查研究

本文介绍了四川大型真菌资源348种,包括药用真菌163种、食用真菌123种、有毒真菌41种、菌根真菌101种等。全国新记录26种。其中冬虫夏草(cordyceps sinensis)、凉山虫草(c.liangshanensis)、松口蘑(Tricholoma matsutake)、银耳(Tremella fuciformis)、短裙竹荪(Dictyophora duplicata)猴头(Hericium erinaceus)、鸡(土从)(Termitomyces albuminosus)、木耳(Auricuiaria auricula)、灵芝(Ganoderma lucidum)等尤为著名。并对四川大型真菌中部分重要种类的分布、资源及用途进行了分析和评价。为合理开发利用及有关教学科研提供了资料。     

Comparative study of mycelial growth and basidiomata formation in seven different species of the edible mushroom genus Hericium

The potential of using several agricultural by-products as supplements of sawdust substrate for the production of edible mushroom Hericium was evaluated using seven Hericium species. All the tested supplements (rice bran, wheat bran, barley bran, Chinese cabbage, egg shell, and soybean powder) were found to be suitable for the mycelial growth of all the tested species. In mycelial growth, soybean powder was the best supplement for Hericium americanum, Hericium coralloides, and Hericium erinaceum while barley bran was the best for Hericium alpestre, Hericium laciniatum, and Hericium erinaceus. For Hericium abietis, rice bran and Chinese cabbage was the best. The possibility of mushroom production on oak sawdust substrate with 20% rice bran supplement was demonstrated with H. coralloides, H. americanum, H. erinaceus, and H. erinaceum which showed 26-70% biological efficiency. Our results also showed that strain selection is important to improve biological efficiency and mushroom yield in Hericium cultivation.     

Studies on the Biology of Macrofungi with Reference to their Cultivation

Nutrition, pH, temperature, moisture, light, and aeration as factors influencing mycelial growth, primordia formation, and development of sporophores of 25 species of basidiomycetous macrofungi were studied, of which 18 species produced normal or nearly normal fruit-bodies in the laboratory. It was found that various agricultural by-products and farm wastes, such as sawdust, wood shavings, fallen leaves, soybean hay, bean pods, corn culms, paddy straw, rice glumes, wild grasses, etc. enriched with 20%–25% by weight of rice or wheatbran, made suitable substrata for most of the lignicolous species investigated with the exception of Tremella fuciformis and Lentinus edodes, and that well fermented dungs of domestic animals, such as horse, cow, pig, sheep, rabbit, and poultry, appeared to be satisfactory substitutes for bran as accelerators for fructification. Yields of carpophores were more or less directly proportional to the percentages of accelerators added to the substratum. The vegetative growth of the majority of species studied took place over a wide pH range of 3.0–8.0, with the optimum at 5–6, while the optimal pH for Ganoderma lucidum, Lentinus edodes, and Hericium erinaceus being about 4.0 and the maximum pH limit for H. erinaceus only 5.4. In Auricularia hispida, the pH range for fructification and that for mycelial growth were almost the same, but in Collybia velutipes and Agaricus bitorquis the pH range for sporogenesis was much more restricted, the optimal being 5–6 and 7–8 respectively. Most of the species investigated produced mycelium within a temperature range of 6–36℃, the optimum being about 24–27℃. However, the optimal temperatures for Auricularia auricula, Poria cocos and Volvariella volvacea var. heimii were found to be 30℃ or more. The temperatures required for sporophore and for primordia formation varied from species to species. While the fruit-bodies of Lentinus edodes and Pleurotus sapidus appeared only after "chilling", temperature fluctuations did not exert any stimu- lating effect on primordia differentiation in some other species. Using the temperature requirement for the initiation of primordia as the criterion, the species studied were grouped into the following categories: (1) high-temperature species, such as Ganoderma lucidum, Pleurotus rhodophyllus, Collybia radicata, Volvariella bombycina, and Volvariella volvacea var. heimii, with their optimum and maximum tem- peratures for primordia initiation exceeding 24℃ and 30℃ respectively; (2) intermediate species, such as Auricularia auricula, Tremella fuciformis, Pleurotus citrinopileatus, Pholiota adiposa, Agaricus bitorquis, and Agaricus rubellus, with their maxima not exceeding 28 ℃ and with the optimum lying within 20–24 ℃; and (3) low-temperature species, such as Auricularia hispida, Hericium erinaceus, H. caput-medusae, Pleurotus sapidus, Lentinus edodes, Collybia velutipes, and Agaricus bisporus, with their maxima not exceeding 24 ℃ and with the optimum under 20 ℃. Thus, under natural climatic conditions of a given region, without artificial heating or cooling, it was deemed practicable to arrange, according to the temperature response of the various species, a series of mushroom crops to be grown in rotation the year round. Moisture was found to have considerable influence on vegetative growth as well as reproduction. Most of the lignicolous species grew well on the sawdust medium, which contained 100%–340% by weight of water. Moisture content between 260%–340% was more favourable for fruiting, and several flushes were secured. At 100% moisture, fruiting was considerably delayed and only one flush of poorly developed carpophores was obtained. Aeration also played an important role in basidiocarp development. Stagnant humid air or CO2-laden atmosphere markedly inhibited pileus development, diminishing the size of pilei and causing morphological abnormalities. Adequate supply of fresh air with 80%–90% relative humidity tended to maintain a normal rate of transpiration, thereby guaranteeing a flourishing crop. In some species, such as Ganoderma lucidum, Tremella fuciformis, Pleurotus sapidus, Pleurotus citrinopileatus, Volvariella bombycina, Hericium erinaceus, Auricularia hispida, Armillaria mucida, and Lentinus edodes, light was found to be absolutely necessary. In these species, primordia formation would not commence until exposed to light. Certain other species, such as Auricularia auricula, Pleurotus rhodophyllus, Collybia velutipes, Pholiota adiposa, and Agaricus bitorquis, were able to fructify in total darkness, though their primordia formation was promoted by light. For the normal development and pig- mentation of fruit-bodies, light appeared to be essential. Morphological abnormalities, such as long slender stipes and thin rudimentary pilei occurred in total darkness. Results of experiments indicated that in most of the species studied, light reduced mycelial growth. The inhibitory effect was mainly due to the blue region (380–540 mμ) of the visible spectrum rather than the red region (570–920 mμ). On the contrary, light at the blue end was shown to be most effective and even essential for fructification; the red end being similar to darkness, evoked almost no response. Having analyzed the results obtained, it seems reasonable to assume that the ontogeny of higher Basidiomycetes consists most probably of four developmental stages, viz., (1) the stage of vegetative growth, (2) the stage of primordia initiation, (3) the stage of sporophore development, and (4) the stage of basidiospore formation, each of which has its own physiological characteristics and metabolic speciality.Hence the requirements of the various stages differ considerably.     

PEG介导的猴头菌遗传转化体系的建立

对猴头菌Hericium erinaceus原生质体制备的各种因素进行比较研究,结果表明,猴头菌原生质体制备的最佳体系为：液体培养5d的猴头菌丝,以0.6mol/L KCl作为稳渗剂,加入含1.0%纤维素酶+1.0%蜗牛酶+1.0%溶壁酶的复合酶,在30℃酶解猴头菌丝3h时,原生质体得率达到3.0×106个/mL。潮霉素敏感性测试表明,猴头菌在PDSA固体培养基上的潮霉素最低筛选浓度为60μg/mL。采用PEG介导的原生质体法,将质粒pBgGI-hph（含有灵芝gpd1-Gl启动子和潮霉素抗性基因hph）转化猴头菌原生质体,经潮霉素初步筛选以及PCR鉴定,表明有4株猴头菌拟转化子的基因组扩增出hph基因;转化子经过多次转接后进行Southern杂交验证,结果表明4个转化子的基因组中均稳定整合了hph抗性基因。