赤芝原生质体分离与再生研究

以赤芝(Ganoderma lucidum)为供试菌种,研究了不同酶解温度、酶解时间、渗透压稳定剂浓度及菌龄、再生培养基等对赤芝菌丝的原生质体制备与再生的影响。结果表明:菌龄为4 d的菌丝,以0.4 mol/m L的甘露醇作渗透压稳定剂配制成浓度为0.02 g/m L的溶壁酶,于p H 5.0、25℃条件下酶解2.0 h,其原生质体数最高可达3.87×107/m L,MYG再生培养基上再生率达到0.016%。     

牛蒡木屑栽培赤芝子实体多糖含量的检测及其配方筛选

为充分利用牛蒡和食药真菌资源,以不同体积比的牛蒡木屑为基质进行赤芝Ganoderma lingzhi栽培。测定不同条件下生长的赤芝子实体多糖含量,从而确定赤芝最优栽培条件。试验结果表明:当栽培基质中牛蒡与木屑体积比1∶4、温度28℃、固液比为20∶0时赤芝子实体多糖含量最高。用此栽培基质,在子实体形成期间,光照条件为暗光、p H=5～6、湿度90%～95%、培育45～60 d,子实体长势最好,形态最佳,其多糖含量可达23.26 mg/g。     

杨木屑栽培蟹味菇关键技术研究

本研究筛选以杨木屑和玉米芯为主的蟹味菇[Hypsizygus marmoreus(Peck) HEBigelow]栽培培养基及优化催蕾条件。经筛选得栽培配方如下:杨木屑35%,玉米芯25%,米糠18%,麸皮18%,白糖1%,石灰1%,石膏1%,过磷酸钙1%,与以棉籽壳为主的常规配方相比,生物转化率略低。采用单因素试验,通过对催蕾影响因素包括:温度,湿度、光照强度和二氧化碳浓度等进行优化研究,获得最佳催蕾条件:15℃下,空气相对湿度为85%～95%时,弱光照条件(光照强度≤50 lx),CO_2浓度控制在020%～025%,利于蟹味菇原基分化,产量和正品率最高。通过栽培培养基及培养条件的筛选研究,证实杨木屑可用于蟹味菇的栽培生产,为其应用提供了具有经济效益的途径。     

赤芝一共生菌发酵过滤液中的新倍半萜酯

从四川野生赤芝(Ganoderma lucidum (Leyss. ex Fr.) Karst.)的一共生菌Calcarisporium arbuscula Preuss的发酵过滤液中分离得到3个新的倍半萜酯类化合物,分别命名为calcarisporin B2 (1)、B3 (2)和B4 (3),通过物理方法和波谱分析确定了它们的结构.     

韩芝液体深层发酵培养基的优化研究

固体培养基上比较了韩芝与树舌的菌丝生长速度,二者平均分别达到0.77 cm.d-1和0.67 cm.d-1;液体培养试验了不同碳氮比、培养基组成对韩芝菌丝生长的影响,得出了菌丝生长较适宜的碳氮比范围在18~24∶1,最适宜的碳氮比在22∶1;对比试验、正交试验得出韩芝液体培养最佳的培养基配方为(g.L-1):葡萄糖10.0、玉米粉20.0、大豆粉20.0、酵母粉5.0;在此条件下,菌丝的平均产量可达23.12 g.L-1。     

灵芝原生质体的分离及其转化

对灵芝(赤芝)进行了原生质体的形成和再生实验,研究了渗透压稳定剂、菌龄、酶解条件对原生质体分离的影响,以及渗透压稳定剂、培养基对原生质体再生的影响;另外还尝试用电穿孔法在灵芝原生质体中进行报告基因(reportgene)β-gal的转化,结果β-gal在菌丝体中有表达。 Abstract:The experiments of protoplasts preparation from Lingzhi (Ganoderma lucidum) and their regeneration were done and the effects of osmotic stabilizer,age of mycelium and lystic conditon on formation and regeneration of protoplasts were studied.The β-geo gene has been transformed into the protoplast by electroporation and the reporter gene β-gal has been expressed in the regenerated hypha.     

响应面法优化以桑枝为主要基质的灵芝栽培配方

目的利用响应面法对以桑枝为主要成分的灵芝栽培培养基配方进行优化。方法以苯酚-硫酸法测定灵芝子实体中的多糖含量为指标,通过Plackett-Burman(PB)实验筛选出影响栽培灵芝子实体多糖含量的主要因素,然后用最陡爬坡试验、中心组合设计和响应面法对影响灵芝多糖含量的显著因素进行优化,利用以栽培灵芝子实体中多糖含量为响应值建立的二次回归方程模型,确定主要影响因素的最佳浓度,得到可大幅度提高灵芝子实体中多糖含量的最佳发酵培养基配方。结果确定了以桑枝为主要基质的优化后栽培培养基配方:65%桑枝屑,25%中药渣,4%玉米粉,1%蔗糖,1%石膏粉,4%麸皮,0.1%KH_2PO_4,0.1%MgSO_4·7H_2O,0.01%VB1。验证实验表明使用优化后的培养基生产的灵芝子实体中的多糖含量高达8.35%,是基础培养基生产的灵芝子实体多糖含量(3.17%)的2.63倍。结论对灵芝栽培培养基配方采用响应面中心组合设计,可以有效地对栽培培养基进行优化。     

冬虫夏草与韩芝液体发酵混合培养特性研究

平板培养基上比较了冬虫夏草、韩芝及树舌的菌丝生长速度和对淀粉的利用情况,结果显示三者的生长速度分别达到1.28、0.64和0.55 cm/d,且韩芝与树舌对淀粉的利用均优于冬虫夏草;混合培养结果得出菌种的最佳菌龄比为虫草:韩芝=3 d:5 d,最佳的接种比例为2:3;正交试验混合发酵最佳的培养基配方(%):葡萄糖1.0、淀粉2.0、黄豆粉2.0、酵母粉0.5,在此条件下,菌丝生物量达到最大值1.97 g/100 mL。     

赤芝子实体的化学成分研究(Ⅱ)

采用硅胶及凝胶柱色谱方法从赤芝(Ganoderma lucidum(Leyss.ex Fr.)Karst.)子实体的乙醇提取物中分离得到10个化合物,通过波谱数据及理化性质分别鉴定为赤芝萜醇A(lucidumol A,1)、麦角甾醇(ergosterol,2)、麦角甾醇棕榈酸酯(ergosteryl palmitate,3)、棕榈酸(palmitic acid,4)、β-谷甾醇(β-sitosterol,5)、硬脂酸(stearic acid,6)、油酸(oleic acid,7)、α-羟基-24-烷酸(α-hydroxytetracosanoic acid,8)、2-羟基-二十四烷酸乙酯(2-hydroxytetracosanoic ethyl ester,9)以及2-羟基-二十四烷酸甲酯(2-hydroxytetracosanoic methyl ester,10)。其中,化合物9~10首次从该属植物中分离得到,4~8首次从该植物中分离得到。     

14个灵芝菌株的酯酶同工酶分析*

对14个灵芝菌株进行拮抗试验和酯酶同工酶聚丙烯酰胺凝胶电泳聚类分析,实验共检出迁移率不同的谱带18条,在50%相似水平上聚合为5类,第1类包括仙芝、大仙823、南韩、园芝6号、甜芝、韩芝;第2类包括赤芝10号、赤芝12号、赤芝、日本红芝;第3类包括紫芝和黑芝;第4类为血芝;第5类为白芝。聚类分析结果与形态观察及拮抗试验结果基本一致。实验还说明酯酶同工酶可作为灵芝良种选育的一种有效标记方法,在菌种鉴定、保藏和管理等方面具有重要作用。     

超高压对灵芝生长及其漆酶合成的影响

研究了超高压处理对灵芝生长及其漆酶合成的影响,试验结果表明:灵芝致死率随处理压力的增大而增加,在150MPa下处理30min获得了一株生物量和漆酶产量都大幅增加的菌株G1502,其最大生物量及酶活分别比出发菌株提高了19.34%和282.67%,发酵时间也缩短了1d。     

超高压对漆酶产生菌的诱变效应研究

对漆酶产生菌灵芝和鸡腿菇进行超高压处理,发现致死率随剂量的增加而增加,在致死率为50%~80%的范围内诱变效果较好。实验获得了2株具有较大应用潜力的灵芝高压突变株G1502和鸡腿菇高压突变株C2003,其中G1502酶活比出发菌株提高了2.83倍,发酵时间缩短了1d;C2003酶活比原菌株提高了2.57倍,发酵时间也缩短了1d。     

荷叶离褶伞优良菌株筛选

以棉籽壳、木屑、玉米芯等为主料设计了10个栽培配方,研究不同配方下7株荷叶离褶伞的菌丝生长和出菇情况。结果表明,各菌株菌丝颜色以雪白色为主,在玉米芯为主料的配方上菌丝体生长优于以棉籽壳、木屑为主料的配方,但均无子实体形成;以棉籽壳为主料有利于子实体的形成,特别是棉子壳添加少量木屑、树叶、腐殖土、发酵料和小麦更利于子实体形成,菌株3001的出菇周期最短（108d）,产量最高（214.80g/袋）,各菌株出菇的顺序为3001、1035、1004和1013。因此,栽培料组分对荷叶离褶伞菌丝生长及子实体形成影响较大。     

Mechanisms of the Anticancer Action of Ganoderma lucidum （Leyss. ex Fr.） Karst.： A New Understanding

Ganoderma lucidum (Leyss. ex Fr.) Karst., a medicinal fungus called “Lingzhi” in China, has been used in traditional Chinese medicine in China for the prevention and treatment of various types of diseases, such as cancer, hepatopathy, arthritis, hypertension, neurasthenia, and chronic hepatitis. It is clear that the anticancer activity of G. lucidum is mainly due to polysaccharides and/or triterpenoids of the fungus. However, until now, the mechanism of the anticancer action of G. lucidum has not been well understood and, previously, the activation of the immune response of the host was widely considered to be the only mechanism by which G. lucidum prevented and/or treated cancer. However, recent studies reviewed in the present paper have shown that the potential mechanisms of anticancer action include not only the activation of the immune response of the host, but also the induction of cell differentiation, the induction of Phase Ⅱ-metabolizing enzymes, the inhibition of angiogenesis, and the inhibition of the expression of the urokinase-type plasminogen activator (uPA) and the uPA receptor in cancer cells. To further elucidate the mechanisms of action of G. lucidum, more in vivo tests and randomized controlled clinical trials should be carried out, and the molecular mechanisms should be studied intensively. Additionally, whether the anticancer compounds in G. lucidum act synergistically or independently should be further studied.     

Mechanisms of the Anticancer Action of Ganoderma lucidum (Leyss. ex Fr.) Karst.: A New Understanding

Ganoderma lucidum (Leyss. ex Fr.) Karst., a medicinal fungus called "Lingzhi" in China, has been used in traditional Chinese medicine in China for the prevention and treatment of various types of diseases, such as cancer, hepatopathy, arthritis, hypertension, neurasthenia, and chronic hepatitis. It is clear that the anticancer activity of G. lucidum is mainly due to polysaccharides and/or triterpenoids of the fungus. However, until now, the mechanism of the anticancer action of G. lucidum has not been well understood and, previously, the activation of the immune response of the host was widely considered to be the only mechanism by which G. lucidum prevented and/or treated cancer. However, recent studies reviewed in the present paper have shown that the potential mechanisms of anticancer action include not only the activation of the immune response of the host, but also the induction of cell differentiation, the induction of Phase Ⅱ-metabolizing enzymes, the inhibition of angiogenesis, and the inhibition of the expression of the urokinase-type plasminogen activator (uPA) and the uPA receptor in cancer cells. To further elucidate the mechanisms of action of G. lucidum, more in vivo tests and randomized controlled clinical trials should be carried out, and the molecular mechanisms should be studied intensively. Additionally, whether the anticancer compounds in G. lucidum act synergistically or independently should be further studied.     

不同基质栽培的猴头菌子实体多糖理化性质和体外免疫活性

【背景】猴头菌多糖是猴头菌(Hericium erinaceus)中主要的活性成分,具有提高免疫力、抗肿瘤、降血糖、抗突变、抗衰老等多种功效。目前对于猴头菌多糖的研究主要集中在结构分析上,有关不同基质栽培对于猴头菌子实体多糖理化性质的影响鲜见报道。【目的】研究不同基质栽培的猴头菌子实体多糖的理化性质和体外免疫活性,筛选出高产优质猴头菌的栽培配方。【方法】采用7种不同配方的基质经工厂化栽培获得不同的猴头菌子实体,对其子实体产量和粗多糖含量进行统计和分析测定,并对7种不同基质栽培获得的猴头菌子实体多糖的分子量分布及单糖组成等理化特征进行分析,比较其对刺激RAW264.7巨噬细胞株释放NO的差异。【结果】综合各项指标表明,配方2 (木屑30%,玉米芯40%,棉籽壳15%,玉米粉2%,麸皮6%,米糠5%,石膏1%,石灰1%)和配方7 (玉米芯39%,棉籽壳10%,麸皮10%,米糠30%,大豆皮5%,甜菜渣4%,碳酸钙2%)栽培获得的猴头菌子实体具有较高的多糖含量和活性。【结论】所选配方适合作为栽培猴头菌深加工原料的培养基质。     

青霉PT95菌固态发酵产生类胡萝卜素的研究

本文对青霉Ｐｅｎｉｃｉｌｌｉｕｍｓｐ．ＰＴ９５菌株在固态发酵条件下菌核内产生类胡萝卜素进行了初步研究。结果表明,在３种固态发酵培养基中,玉米粉培养基（ＳＭＡ）比麸皮２基和棉籽壳培养基更适合于ＰＴ９５菌株固态发酵产生类胡萝卜素。为了增加菌核干重和提高类胡萝卜素产率,ＳＭＡ中需要添加氮源、碳源和植物油。在所度的各种氮、碳源中,以硝酸钠和麦芽糖效果最佳。通过我试验确定了在培养基盐溶液中添加硝酸钠３ｇ／Ｌ     

富钙灵芝的深层培养

目的:研究灵芝深层发酵过程中钙对灵芝生长及钙的富集作用的影响。方法:用重氮化法将ABA分别与牛血清白蛋白(BSA)、卵清白蛋白(OVA)连结,得到ABA的免疫抗原和包被抗原,并采用紫外全波长扫描和SDS-PAGE鉴定合成的抗原;以经过鉴定的抗原免疫白兔,制备出ABA的多克隆抗体;采用间接酶联免疫法对抗血清进行效价检测;通过离子交换层析法获得纯化的抗体。结果:当培养基中硝酸钙的添加量达到600mg/100g时,灵芝生长和富集钙的效果最好;添加量超过800mg/100g时,灵芝富集钙的效果明显降低。富钙灵芝中有机钙的含量约占总钙量的97.30%。结论:在培养基中添加低浓度的钙,对灵芝的生长和钙的富集有促进作用,为开发新型有机钙提供了依据。     

Evaluation of edible mushroom Oudemansiella canarii cultivation on different lignocellulosic substrates

In this study, the mycelial growth rate, mycelial colonization time, yield, and biological efficiency of the edible mushroom Oudemansiella canarii were determined, and the effects of different substrate combinations on productivity, chemical contents and amino acids were evaluated. Lignocellulosic wastes, such as cottonseed hull, sawdust, corncob, and their combinations supplemented with 18% wheat bran and 2% lime, were used for the cultivation of O. canarii. The biological efficiency (BE) and essential amino acid content of treatment T1, which consisted of 80% cottonseed hull, were the highest among all the tested treatments. Mixtures that included sawdust, such as treatments T2 (80% sawdust), T4 (40% sawdust + 40% cottonseed hull), and T6 (40% sawdust + 40% corncob), exhibited lower yield and BE. Corncob was good for O. canarii production in terms of yield and BE, whereas the mycelial growth rate and colonization time were lower compared to those on other substrates. Comparing the BE, essential amino acids, and other traits of the six treatments, treatment T1 (80% cottonseed hull) was the best formula for O. canarii cultivation and should be extended in the future.     

Solid-state fermentation of cornmeal with the basidiomycete Ganoderma lucidum for degrading starch and upgrading nutritional value

AIMS: The objective of this research was to study the ability of the basidiomycete Ganoderma lucidum to degrade starch and upgrade nutritional value of cornmeal during solid-state fermentation (SSF). METHODS AND RESULTS: On the basal medium that consisted of cornmeal and salt solution, alpha-amylase activity of G. lucidum reached its maximum value of 267 U g(-1) of culture on day 20 after inoculation. Prolongation of fermentation time from 10 to 25 days increased significantly the degradation rate of starch and ergosterol yield (a kind of physiologically active substances of G. lucidum, also as an indicator of mycelial biomass) (P < 0.01). Supplementation of glucose, sucrose or maltose to the basal medium also caused a significant increase in either the degradation rate of starch or the ergosterol yield as compared with control (P < 0.01). Among five kinds of nitrogen sources supplemented, yeast extract, casamino acid and peptone were more effective than (NH4)2SO4 and NH4NO3, and yeast extract gave the highest degradation rate of starch and ergosterol yield, followed by peptone. Through orthogonal experiments, the theoretical optimum culture medium for SSF of this fungus was the following: 100 g cornmeal, ground to 30-mesh powder, moistened with 67 ml of nutrient salt solution supplemented with 3 g yeast extract and 7.5 g glucose per litre. CONCLUSIONS: Under the optimum culture condition, the degradation rate of starch reached its maximum values of 70.4%; the starch content of the fermented product decreased from 64.5 to 25.3%, while the reducing sugar content increased from 4.2 to 20.6%. SSF also produced a significant increase (P < 0.01) from 11.0 to 16.5% in protein content. SIGNIFICANCE AND IMPACT OF THE STUDY: After SSF by G. lucidum, the digesting and absorbing ratio of cornmeal was strikingly increased and some active substances originated from G. lucidum remained in the fermented product. This implied that cornmeal could be processed into many kinds of special functional foods by SSF of G. lucidum.