三隔镰刀菌大米培养物产生二苯甲醇的研究

<正> T-2毒素是镰刀菌属三隔镰刀菌Fusarium tricinctum(Corda)Sacc.的一种次生代谢产物,隶属单端孢霉烯簇毒素。发霉大米中毒症及豆荚中毒症等均与单端孢霉烯簇毒素有关。张树荣等从真菌培养物M-20中提取T-2毒素成功,我们对其     

青岛崂山的重寄生菌白粉寄生孢及其寄主多样性

2005－2007年从青岛崂山的16科30属35种植物上采集了143份白粉菌样品,经形态鉴定分属于7属22种。其中40份样品中检测到白粉寄生孢Ampelomyces quisqualis,其寄主植物属于8科14属15种,其寄主真菌属于7属10种白粉菌。统计分析发现,多孢穆氏节丝壳上白粉寄生孢的发生率和重寄生强度均为最高;群落组成分析发现,棕丝单囊壳为白粉寄生孢寄主真菌中的优势种,其重要值为89.22%;菊科为白粉寄生孢寄主植物中的优势科,其重要值为97.85%。     

中国枝孢菌属的分类研究Ⅻ.露珠草上一新种(英文)

报道寄生在柳叶菜科（Onagraceae）南方露珠草（CircaeamollisSieb.etZuu．）上枝孢菌属一新种：露珠草枝孢CladosporiumcircaeaeY．QinetZ．Y．Zhangsp．nov。枝孢菌属真菌在柳叶菜科是首次报道。文中为新种提供了拉丁文描述和附图。模式标本保藏于云南农业大学真菌标本室（MHYAU）。     

荚孢腔属真菌次生代谢产物的研究进展

荚孢腔属(Sporormiella)真菌,属于座囊菌纲格孢腔菌目荚孢腔菌科(Sporormiaceae),种类较多,目前已收载了87种。研究发现,该属真菌的次生代谢产物资源丰富,迄今已报道的新化合物就有近50个,包括生物碱及含氮类、聚酮类、萜及甾体等化合物,其中已发现一些结构新颖、生物活性独特的化合物,为药物的研发提供了潜在的先导化合物,具有很好的研究价值。然而,目前对该属真菌的次生代谢产物研究的报道还较少,为使该属真菌资源能引起人们的关注和更多认识,本文对荚孢腔属真菌次生代谢产物的化学结构及其生物活性的研究进展进行了较为全面的综述。     

新疆的刺球座属、穴壳属及普氏腔孢属

本文报道了新疆腔菌纲座囊菌目刺球座属(Lasiobotrys)、穴壳属(Dothiora)和普氏腔孢属(Plowrightia)的六种子囊菌,即:忍冬刺球座菌(L.loniccrae)、花楸穴壳菌 (D.sorbi)及其无性阶段花楸疡壳孢(Dothichiza sorbi)、茶蔗子普氏腔孢菌(P.ribesia)、小檗普氏腔孢菌(P.berberidiJ)、沙棘普氏腔孢菌(P.hippophaeos)及雕刻普氏腔孢菌(P.insculpta)。这三个属的真菌在我国均未报道过,为我国新纪录属(种)。标本均采于新疆,保存于新疆八一农学院植保系真菌标本室(HMAAC)。     

四绺孢球腔菌科真菌地理分布及其生态学功能

根据ITS、LSU、rpb2、tef1和tub2多基因系统发育分析,将云南禾本科植物格孢腔菌目的7株内生真菌鉴定归属于格孢腔菌目Pleosporales四绺孢球腔菌科Tetraplosphaeriaceae的四绺孢属Tetraploa和假四绺孢属Pseudotetraploa以及该目下的一个未定属genera incertae sedis。羧甲基纤维素钠培养基和愈创木酚培养基筛选发现四绺孢球腔菌科的2个菌株具有较强的纤维素酶和漆酶活性,而这个未定属的菌株仅具有较弱的纤维素酶活性、无漆酶活性,表明格孢腔菌目的2个内生真菌类群的纤维素和木质素降解能力不同。多重对应分析发现四绺孢球腔菌科真菌的属与寄主、分离来源和地理位置有关联,其中四绺孢属和假四绺孢属可在活的健康植物作为内生真菌存活,并在植物凋落物和土壤中分离得到,推测四绺孢属和假四绺孢属两属为内生和腐生双生态位真菌。因此,进一步深入探究四绺孢球腔菌科内生真菌参与的禾本科植物凋落物的分解将深化我们对禾本科植物内生真菌多样性和生态学功能的认识。     

嗜角蛋白真菌的界定、研究方法及其应用价值

嗜角蛋白真菌(keratinophilic fungi)是指能降解角蛋白并能利用其作为唯一营养源的真菌类群,大部分隶属于爪甲团囊菌目(Onygenales)、散囊菌目(Eurotiales)和肉座菌目(Hypocreales).由于该类群分布广、种类多、营养方式多样,其研究方法和技术手段也在不断进步,加深了其研究的深度...     

引起泡花树属叶斑的尾孢菌属一新种

报道寄生在清风藤科 Sabiaceae 泡花树属 Meliosma sp. 植物上尾孢菌属的一个新种:泡花树尾孢 Cercospora meliosmae。尾孢菌属真菌在清风藤科尚属首次报道。文中为新种提供了拉丁文简介、图及英文描述,模式标本保存在中国科学院微生物研究所菌物标本馆(HMAS)。     

中国大单孢属真菌一新纪录种

大单孢属(Aplosporella)隶属于真菌界(Fungi),子囊菌门(Ascomycota),盘菌亚门(Pezizomycotina),座囊菌纲(Dothideomycetes),葡萄座腔菌目(Botryosphaeriales),葡萄座腔菌科(Botryosphaeriace-ae)的一个无性态属。大单孢属成员常常生于植物的小枝条上,引起植物小枝条枯死。该属包含多个异源种,这些异源种需要进一步归到其他相关属里去[1]。     

中国热带地区的半知菌Ⅰ.云南尾孢菌属及其近似属的种

本文报道了云南热带地区尾孢菌属及其近似属真菌４５种,其中有一一个新种：番木瓜短胖孢寄生在番木瓜科番木瓜叶上。短胖孢属真菌在番木瓜科植物上是首次报道。文中为新种提供了拉丁文简介,描述并附图。模式标本保藏在中国科学院微生物研究所真菌标本室。     

中国金孢属一新记录种

Fermentation broth of endophytic fungus Trichoderma taxi ZJUF0986 has high antifungal activity to the common 15 species of phytopathogenic fungi. Three bioactive metabolites I, II and III were obtained by extracted with petroleum ether, ethyl acetate and n-butanol and subsequent silica gel column chromatography and preparative HPLC. Among them, compound I, the main metabolite, has strong broad-spectrum antifungal activity, especially to Botrytis cinerea, Rhizoctonia solani and Sclerotinia sclerotiorum with IC50 1.06, 1.08 and 1.13 mg/L, respectively.     

紫杉木霉ZJUF0986抗真菌活性代谢产物分离提取及其活性研究

南方红豆杉Taxus chinensis var.maimi内生真菌紫杉木霉Trichoderma taxi菌株ZJUF0986发酵液对15种常见植物病原真菌具有很强的抑菌活性;用石油醚、乙酸乙酯和正丁醇对发酵液进行活性物质的提取分离,并通过硅胶柱层析和制备HPLC纯化,得到3个活性代谢产物Ⅰ、Ⅱ和Ⅲ,其中主要活性代谢产物Ⅰ具有广谱高效抑制植物病原真菌的特点,特别是对灰葡萄孢Botrytis cinema、立桔丝核菌Rhizoctonia solani、核盘菌Sclerotinia sclerotiorum具有很强的抑菌活性,其IC50为1.1mg/L.     

洋葱伯克霍尔德菌CF_66抗菌物质的分离纯化及性质的研究

洋葱伯克霍尔德菌CF-66能够抑制立枯丝核菌等若干植物病原菌和其它一些真菌的生长。CF-66菌发酵液的粗提液通过Sephadex-75pg、Sephacryl S-100柱层析分离纯化,获得抗菌物质CF66I。此抗菌物质耐热性强,耐碱,但在强酸性条件下不稳定。低浓度有机溶剂的存在有利于抑菌活性的提高。对其结构的研究表明CF66I是以(CH2CH2O)n为主要单元结构并带有酰氨键的化合物。     

Alkaline protease inhibitor: a novel class of antifungal proteins against phytopathogenic fungi.

A Streptomyces sp., which produces an alkaline protease inhibitor (API) exhibiting antifungal activity has been isolated from soil. The protein has been purified to homogeneity. The molecular characterization has revealed that it is a dimer (M(r) 28 kDa) with five disulphide linkages and has a pI of 3.8. API is a competitive type of inhibitor with a K(i) value of 2.5 x 10(-9) M. The inhibitor is stable over a pH range of 6 to 12 and a temperature range of 40 to 95 degrees C. API exhibits antifungal activity (in vitro) against phytopathogenic fungi such as Fusarium, Alternaria, and Rhizoctonia and also against Trichoderma, a saprophytic fungus. The antifungal activity of API appears to be associated with its ability to inhibit the fungal serine alkaline protease(s), which is indispensable for its growth. Retardation of the rate of fungal spore germination, as well as hyphal extention, was observed in the presence of API. Both the protease inhibitory and the antifungal activity were abolished on treatment of API with DTT (5 mM), suggestive of a common site for both the activities. This is the first report on API as a potential biocontrol agent against phytopathogenic fungi.     

Expression and functional analysis of two osmotin (PR5) isoforms with differential antifungal activity from Piper colubrinum: prediction of structure-function relationship by bioinformatics approach

Osmotin, a pathogenesis-related antifungal protein, is relevant in induced plant immunity and belongs to the thaumatin-like group of proteins (TLPs). This article describes comparative structural and functional analysis of the two osmotin isoforms cloned from Phytophthora-resistant wild Piper colubrinum. The two isoforms differ mainly by an internal deletion of 50 amino acid residues which separates them into two size categories (16.4 kDa-PcOSM1 and 21.5 kDa-PcOSM2) with pI values 5.6 and 8.3, respectively. Recombinant proteins were expressed in E. coli and antifungal activity assays of the purified proteins demonstrated significant inhibitory activity of the larger osmotin isoform (PcOSM2) on Phytophthora capsici and Fusarium oxysporum, and a markedly reduced antifungal potential of the smaller isoform (PcOSM1). Homology modelling of the proteins indicated structural alterations in their three-dimensional architecture. Tertiary structure of PcOSM2 conformed to the known structure of osmotin, with domain I comprising of 12 β-sheets, an α-helical domain II and a domain III composed of 2 β-sheets. PcOSM1 (smaller isoform) exhibited a distorted, indistinguishable domain III and loss of 4 β-sheets in domain I. Interestingly, an interdomain acidic cleft between domains I and II, containing an optimally placed endoglucanase catalytic pair composed of Glu-Asp residues, which is characteristic of antifungal PR5 proteins, was present in both isoforms. It is well accepted that the presence of an acidic cleft correlates with antifungal activity due to the presence of endoglucanase catalytic property, and hence the present observation of significantly reduced antifungal capacity of PcOSM1 despite the presence of a strong acidic cleft, is suggestive of the possible roles played by other structural features like domain I or/and III, in deciding the antifungal potential of osmotin.     

Combinatorial approach to lead optimization of a novel hexapeptide with antifungal activity

Three sets of sublibraries of an antifungal lead peptide His-D-Trp-D-Phe-Phe-D-Phe-Lys-NH2 (I) have been prepared by introducing variations at positions 1, 4 and 6. They were screened for their antifungal activity against C. albicans and C. neoformans in order to quantify inhibition at each step of the hexapeptide sublibrary iteration. The studies led to the identification of Arg-D-Trp-D-Phe-Ile-D-Phe-His-NH2 as a novel hexapeptide with potent antifungal activity against both C. albicans and C. neoformans.     

Crystal structure of tobacco PR-5d protein at 1.8 A resolution reveals a conserved acidic cleft structure in antifungal thaumatin-like proteins

The crystal structure of tobacco PR-5d, an antifungal thaumatin-like protein isolated from cultured tobacco cells, was determined at the resolution of 1.8 A. The structure consists of 208 amino acid residues and 89 water molecules with a crystallographic R-factor of 0.169. The model has good stereochemistry, with respective root-mean-square deviations from the ideal values for bond and angle distances of 0.007 A and 1.542 degrees. Of the homologous PR-5 proteins, only those with antifungal activity had a common motif, a negatively charged surface cleft. This cleft is at the boundary between domains I and II, with a bottom part consisting of a three-stranded antiparallel beta-sheet in domain I. The acidic residues located in the hollow of the cleft form the beta-sheet region. Sequence and secondary structure analyses showed that the amino acid residues comprising the acidic cleft of PR-5d are conserved among other antifungal PR-5 proteins. This is the first report on the high-resolution crystal structure of an antifungal PR-5 protein. This structure provides insight into the function of pathogenesis-related proteins.     

Development of a plasma high-performance liquid chromatographic assay for LY303366, a lipopeptide antifungal agent, and its application in a dog pharmacokinetic study

An HPLC assay for plasma analysis of LY303366 (I), a semi-synthetic lipopeptide antifungal related to echinocandin B (ECB), was developed to support the selection and subsequent preclinical development of I. The method involved extraction of I from plasma with the aid of solid-phase extraction (SPE) cartidges followed by reversed-phase HPLC with UV detection at 300 nm. The method is simple, selective and is applicable to dog, rat, mouse and rabbit plasma. Validation studies using dog plasma showed that the values obtained for parameters of linearity, precision and accuracy were within acceptable limits. Based on analysis of 0.3 ml of plasma, the lower limit of quantitation was 20 ng/ml. The method has been successfully applied to determine the pharmacokinetic parameters of I in the dog following intravenous (i.v.) and oral administration. Compared to first generation ECB antifungal agents, the results of the i.v. dog study indicated a 50% reduction in clearance of the drug from plasma (0.1 l/h/kg) and an 18-fold increase in the volume of distribution at steady state (1.8 l/kg). When administered orally, compound I had an absolute bioavailability of 9%; however, plasma levels remained above the MIC for C. albicans (0.005 μg/ml) through 48 h. Given the excellent potency of I and its broad spectrum of activity relative to first generation ECB antifungal agents, the assay results for I indicate the potential for its use as a broad spectrum i.v. and oral antifungal agent.     

Characterization and antifungal activity of gazyumaru (Ficus microcarpa) latex chitinases: both the chitin-binding and the antifungal activities of class I chitinase are reinforced with increasing ionic strength

Three chitinases, designated gazyumaru latex chitinase (GLx Chi)-A, -B, and -C, were purified from the latex of gazyumaru (Ficus microcarpa). GLx Chi-A,-B, and -C are an acidic class III (33 kDa, pI 4.0), a basic class I (32 kDa, pI 9.3), and a basic class II chitinase (27 kDa, pI > 10) respectively. GLx Chi-A did not exhibit any antifungal activity. At low ionic strength, GLx Chi-C exhibited strong antifungal activity, to a similar extent as GLx Chi-B. The antifungal activity of GLx Chi-C became weaker with increasing ionic strength, whereas that of GLx Chi-B became slightly stronger. GLx Chi-B and -C bound to the fungal cell-walls at low ionic strength, and then GLx Chi-C was dissociated from them by an escalation of ionic strength, but this was not the case for GLx Chi-B. The chitin-binding activity of GLx Chi-B was enhanced by increasing ionic strength. These results suggest that the chitin-binding domain of basic class I chitinase binds to the chitin in fungal cell walls by hydrophobic interaction and assists the antifungal action of the chitinase.     

Identification of novel antifungal nonapeptides through the screening of combinatorial peptide libraries based on a hexapeptide motif

Four sets of mixture based nonapeptide libraries derived from an antifungal hexapeptide pharmacophore Arg-D-Trp-D-Phe-Ile-D-Phe-His-NH(2) (II) have been synthesized. The three C-terminal positions 7, 8 and 9 were subject to randomization using 19 genetically coded amino acids. They were then screened for their antifungal activity against Candida albicans and Cryptococcus neoformans in order to quantify inhibition at each step of the nonapeptide sublibrary deconvolution. The studies led to the identification of several novel nonapeptides with potent antifungal activity. Two of the nonapeptides exhibited approximately 17-fold increase in the activity in comparison to the lead hexapeptide motif His-D-Trp-D-Phe-Phe-D-Phe-Lys-NH(2) (I) against C. albicans.