一种含有单链RNA的香菇球状病毒

从生长不正常的香菇(Lentinus edodes(Berk.)Sing)菌株中分离到一种等轴对称含单链RNA的病毒颗粒。病毒颗粒在电镜下直径为33～34nm,在SDS-聚丙烯酰胺凝胶电泳中病毒外壳蛋白分子量为22000道尔顿。病毒核酸径DNase1和SI酶解试验及热变性紫外吸收曲线试验证明为单链RNA,在1.5％的琼脂糖凝胶电泳中,病毒核酸呈现一条带,分子量为2.38×10~6道尔顿。     

香菇子实体蛋白我糖的分离纯组成结构分析

香菇〔Lentinus edodes（Berk.）Sing.〕子实体经热水提取,乙醇沉淀,得多糖粗品（Le）。Le脱游离蛋白,脱色,经DEAE－纤维素柱及纤维素凝胶柱层析分离酏化和到Le－1、Le－2和 Le－33个级分。经聚丙烯酰胺凝胶电泳和SepharoseCL－6B柱层析鉴定为分子量分布均一的蛋白多糖。凝胶渗透色谱法测得Le－1、Le－2和 Le－3分子量分别为954000,90000和14000。3个级分均由阿拉伯糖、木糖、甘露糖、半乳糖、葡萄糖和葡萄糖醛酸组成,气相色谱测得Le－1的阿拉伯糖、木糖、甘露糖、半乳糖、葡萄糖的摩尔比为0．39：0．46：1．00：0．93：14．13;Le－2为0．19：0．41：1．00：0．93：10．72;Le－3为0．31：0．47：1．00：1．15：8．92。Le－1、Le－2和 Le－3葡萄糖 醛酸含量（％）分别为24．10、34．77和40．05,蛋白质含量（％）分别为2．01、7．28和25．31,红外扫描确定Le－1和Le－2 的糖苷键为a型,Le－3为β型。     

Characterization of an extracellular glycolipid from Lentinus edodes (Berk.) Sing [Lentinula edodes (Berk.) Pegler]

Submerged mycelium of a xylotrophic basidiomycete Lentinus edodes produces an extracellular glycolipid, S3, associated with a lectin. Galactose glycan residue, as well as the lipid pool composition, which includes nonhydroxylated short-chain fatty acids, is uncommon for basidiomycetes. The glycolipid consists of D-galactopyranose (15% of S3 contains galactose sulfate) acylated by octadecanoic and nonadecanoic fatty acid residues (28 and 72%, respectively). The glycolipid structure and composition are confirmed by physicochemical analysis. The glycolipid is assumed to be a regulator of lectin activity.     

Hemagglutinating activity of the fungusLentinus edodes (Berk.) sing [Lentinus edodes (Berk.) Pegler]

The hemagglutinating (HA) activity of the submerged mycelium and the culture liquid (CL) of four strains ofLentinus edodes was studied. The HA activity of the CLs proved to be much higher than that of mycelia. The carbohydrate specificity of fungal agglutinating factors was determined. HA activity was investigated as a function of the inoculum size, cultivation temperature, and culture age. The agglutinating activity of different morphogenetic structures ofL. edodes F-249, including mycelium, brown mycelial mat (MM), primordia, and fruiting bodies, was studied. MM was found to possess the maximum HA activity, which can be explained by the possible involvement of agglutinins in the formation of MM, which is composed of glued hyphae.     

香菇蛋白质氨基酸的分析

分析结果表明,香菇的菌柄与菌盖中蛋白质氨基酸均为18种,缺少谷氨酰胺(Gln)和天冬酰胺(Asn),其中,含量最高的是谷氨酸(菌柄为11.03 mg/gDW,菌盖为12.57 mg/gDW)。菌盖与菌柄均含必需氨基酸、半必需氨基酸10种,其中精氨酸含量(菌柄为10.73 mg/gDW,菌盖为11.84 mg/gDW)。必需氨基酸的含量十分接近于非必需氨基酸的含量(比率为1.00∶1.04),无论总氨基酸含量还是必需氨基酸含量,菌盖中的皆略高于菌柄中的。     

Relationship between the Molecular Structure of the Nitrogen Source and the Activity of the Extracellular Lectins of Lentinus edodes (Berk.) Sing [Lentinula edodes (Berk.) Pegler] upon Submerged Cultivation

The activity of the extracellular lectins of Lentinus edodes (Berk.) Sing [Lentinula edodes (Berk.) Pegler] and the formation of a pigmented mycelial film by this fungus upon submerged cultivation in a synthetic medium were found to depend on the presence of some amino acids (particularly, asparagine) and Ca²⁺ and Mn²⁺ ions in the medium. Quantum-chemical calculations suggest that the different character of the interaction of amino acids with the aforementioned ions is due to differences in the hydrophobicity of the amino acids rather than to differences in the electron structure of the amino acid zwitterions.     

香菇栽培料新工艺

在国内香菇栽培料常规配方基础上,设计了添加2.5mL盐酸/kg取代常规配方中1%蔗糖的新工艺,经测定,2种栽培料灭菌后还原糖与蔗糖浓度、发菌期与子实体产量基本一致,新配方栽培料灭菌后pH值为5.6,是香菇菌丝生长最适pH值。新配方工艺可降低生产成本10%左右。     

香菇菌丝体多糖的分离鉴定与免疫功能研究

从香菇（Ｌｅｎｔｉｎｕｓｅｄｏｄｅｓ）菌丝体提取得到的粗多糖,经ＤＥ５２柱层析,得到均一的多糖成分,命名为ＬＥ。其平均分子量为５．０８×１０５,ＬＥ经红外（ＩＲ）和紫外（ＵＶ）光谱分析,为多糖蛋白质复合体,其中糖含量为９４．２％,蛋白质含量为５．８％。完全酸水解表明糖组成为单一的葡萄糖,蛋白质组成主要为Ａｌａ等１６种天然氨基酸。ＬＥ经完全甲基化、酸水解、乙酰化、薄层层析（ＴＬＣ）和［１Ｈ］核磁共振（［１Ｈ］ＮＭＲ）分析,确定其多糖部分的基本结构为１→３连接的葡聚糖,含有部分１→６侧链。此外,用反转录聚合酶链式反应（ＲＴＰＣＲ）和生物测定法分别研究了健康人外周血单核细胞中白细胞介素２（ＩＬ２）和肿瘤坏死因子（ＴＮＦα）的基因表达和活性,结果发现ＩＬ２和ＴＮＦα的基因表达和活性均高于空白对照组,这表明ＬＥ可诱导某些免疫反应。     

香菇双单杂交后代不同发育阶段酯酶同工酶研究

选用香菇野生株分别与栽培株及杂交株进行双单杂交 ,得到 8个杂交后代 ,且具有结实能力 ,分别对液培 2 0d杂交后代菌丝与液培原基酯酶同工酶进行了比较研究。结果表明 ,液培 2 0d菌丝菌株间酯酶同工酶酶谱显示出多样性 ,可以作为鉴定菌株的辅助遗传标记 ,而原基菌株间酯酶同工酶酶谱谱带较少 ,呈现趋同效应 ,不宜作为鉴定香菇菌株的辅助遗传标记。     

Effect of nutrient nitrogen and manganese on manganese peroxidase and laccase production by Lentinula (Lentinus) edodes

Abstract Lentinula (Lentinus) edodes , strain LS4, produces manganese-dependent peroxidase (MnP) and laccase, but not lignin peroxidase, when grown on a defined medium with glucose as sole carbon source. MnP production is suppressed by nitrogen whereas highest levels of laccase were observed when the fungus was grown under high nitrogen (26 mM) conditions. Both the titre and time of appearance of MnP were affected by the concentration of Mn in the culture medium with highest enzyme levels recorded in cultures supplemented with 1.1 ppm Mn. Purified MnP from L. edodes LS4 has an apparent M _r of 59000 and a p I of 5.6, and differs in several respects from a MnP isolated from L. edodes grown on a commercial wood substrate.     

Physicochemical properties and acute toxicity studies of a lectin from the saline extract of the fruiting bodies of the shiitake mushroom,Lentinula edodes (Berk)

A lectin (LEL) was isolated from the fresh fruiting bodies of the shiitake mushroom Lentinula edodes by a combination of gel filtration chromatography on Sephadex G-150 and affinity chromatography on an N-acetyl-Dgalactosamine-Sepharose 4B column. Its molecular mass, as determined by gel filtration, was estimated to be 71, 000 Daltons and its structure is homotetrameric with subunit molecular weight of approximately 18,000 Daltons. LEL agglutinated non-specifically red blood cells from the human ABO system as well as rabbit erythrocytes and in haemagglutination inhibition assays, exhibited sugar-binding specificity toward N-acetyl-D-galactosamine. EDTA had no inhibitory effect on its haemagglutinating activity, which was stable up to 70°C and was not affected by changes in pH. The lectin had no covalently-linked carbohydrate and amino acid composition analysis revealed that it contained 124 amino acid residues and was rich in tyrosine, proline, phenylalanine, arginine, glutamic acid and cysteine. LEL did not cause mortality neither was it observed to alter the morphology of key organs when administered intraperitoneally at concentrations up to 10,000 mg kg^-1 body weight of mice.     

Characterization of an extracellular glycolipid from <Emphasis Type="Italic">Lentinus edodes</Emphasis> (Berk.) Sing [<Emphasis Type="Italic">Lentinula edodes</Emphasis> (Berk.) Pegler]

Submerged mycelium of a xylotrophic basidiomycete Lentinus edodes produces an extracellular glycolipid, S3, associated with a lectin. Galactose glycan residue, as well as the lipid pool composition, which includes nonhydroxylated short-chain fatty acids, is uncommon for basidiomycetes. The glycolipid consists of D-galactopyranose (15% of S3 contains galactose sulfate) acylated by octadecanoic and nonadecanoic fatty acid residues (28 and 72%, respectively). The glycolipid structure and composition are confirmed by physicochemical analysis. The glycolipid is assumed to be a regulator of lectin activity.     

Isolation and characterization of an immunoenhancing glucan from alkaline extract of an edible mushroom, Lentinus squarrosulus (Mont.) Singer

A water-soluble glucan, isolated from the alkaline extract of the fruit bodies of an edible mushroom, Lentinus squarrosulus (Mont.) Singer was found to consist of (1→3,6)-linked, (1→3)-linked, (1→6)-linked, and terminal β-d-glucopyranosyl moieties in a relative proportion of approximately 1:2:1:1. This polysaccharide showed optimum activation of macrophages as well as splenocytes and thymocytes at 10 μg/mL. Structural investigation was carried out using sugar analysis, methylation analysis, periodate oxidation study, and NMR experiments (¹H, ¹³C, DEPT-135, DQF-COSY, TOCSY, NOESY, ROESY, HMQC, and HMBC). On the basis of above-mentioned experiments, the structure of the repeating unit of the polysaccharide was established as:     

香菇C_(91-3)转录本Unigene14872基因Pkinase结构域的克隆表达及其生物信息学分析

旨在克隆香菇C91-3转录本Unigene 14872基因中的Pkinase结构域,并对其进行生物信息学分析。提取香菇C91-3菌丝体中总RNA,根据转录组测序结果,用3'-Full RACE、5'-Full RACE方法获得Unigene 14872基因全长。用NCBI对其进行生物信息学分析,预测其具有丝氨酸/苏氨酸蛋白激酶(Pkinase)结构域。设计引物PCR扩增Pkinase结构域,然后克隆至pET32a(+)载体,再热转化至JM109中保存并进行质粒测序。重组质粒pET32a(+)-Pkinase构建成功,并成功诱导表达重组蛋白,初步鉴定该蛋白具有抗肿瘤活性,为进一步研究重组蛋白的活性及作用机制奠定基础。     

Solution properties of an alpha-(1-->3)-D-glucan from Lentinus edodes and its sulfated derivatives

A water-insoluble alpha-(1-->3)-D-glucan (A) from Lentinus edodes was fractionated into 13 fractions in dimethyl sulfoxide containing 0.25 M lithium chloride (0.25 M LiCl-Me(2)SO). Five fractions were treated with sulfur trioxide-pyridine complex at 25 degrees C to synthesize water-soluble sulfated derivatives (S-A). The weight-average molecular weights, M(w), and intrinsic viscosities [eta], of the samples A and S-A were determined by multi-angler laser light scattering (MALLS), and viscosity. The M(w) dependence of [eta] and of the radius of gyration (z)(1/2), was found to be represented approximately by [eta]=4.9 x 10(-2) M(w)(0.67) (cm(3) g(-1)), and (z)(1/2)=4.8 x 10(-2) M(w)(0.54) (nm) for the alpha-glucan in 0.25 M LiCl-Me(2)SO in the M(w) range from 7.24 x 10(4) to 4.21 x 10(5), and by [eta]=6.8 x 10(-4) M(w) 1.06 (cm(3) g(-1)), and (z)(1/2)=9.4 x 10(-4) M(w)(0.92) (nm) for the sulfated alpha-glucan in aqueous 0.5 M NaCl in the M(w) range from 5.92 x 10(4) to 1.42 x 10(5) at 25 degrees C. The results indicate that the alpha-(1-->3)-D-glucan exists as a flexible chain in 0.25 M LiCl-Me(2)SO, and its sulfated derivative in 0.5 M NaCl aqueous has stiffer chains than the original. (13)C NMR indicated that intramolecular hydrogen bonding occurred in the sulfated alpha-glucan, causing the observed chain stiffness.     

不同产地冬虫夏草及不同厂家虫草发酵菌丝的可溶性蛋白比较

冬虫夏草〔Ｃｏｒｄｙｃｅｐｓｓｉｎｅｎｓｉｓ (Ｂｅｒｋ .)Ｓａｃｃ.〕为我国滋补名贵出口中药 ,主产于青海、西藏、四川、云南、甘肃等省。因野生资源有限 ,严重脱销。为解决药源问题 ,近年来我国已开发出了虫草菌丝体发酵产品[1] 。由于虫草系真菌寄生在昆虫体上的复合体     

Cryopreservation of shoot tips from in vitro plants of sweet potato [Ipomoea batatas (L.) Lam.] by vitrification

 Routine cryopreservation of shoot tips from sweet potato [Ipomoea batatas (L.) Lam] has been hampered by their survival variability after cryogenic exposure. We examined the effects of light conditions on stock plants, sucrose preculture and cryoprotectant loading on survival after vitrification using PVS2 solution. The survival of vitrified sweet potato shoot tips cooled to approximately –208  °C was increased by preculturing with 0.3 M sucrose for 24 h at 22  °C. Survival was also enhanced by excising shoot tips immediately after the 8-h dark photoperiod. The best survival after cryogenic exposure was obtained using 2 M glycerol +0.4 M sucrose for 1 h at 22  °C followed by dehydration with PVS2 for 16 min at 22  °C. Rapid cooling was used and achieved by the immersion of foil strips into partially solidified nitrogen. Successfully vitrified and warmed shoot tips directly developed shoots on a medium containing 1 μM NAA, 0.5 μM BA and 0.1 μM kinetin with only minimum callus formation. Shoot formation occurred in all surviving shoot tips. This procedure shows promise for cryopreserving sweet potato shoot tips. Received: 2 March 1999 / Revision received: 21 September 1999 / Accepted: 29 September 1999