Microbiological 1 alpha-hydroxylation of norethisterone.

Acremonium strictum and Acremonium kiliense converted norethisterone predominantly into its 1α-hydroxyl derivative. Chemical and spectroscopic /UV, IR, PMR, MS, ORD, CD/ methods were used in establishing the structure and stereochemistry of the product. The 1α-hydroxyl group is shown to be axially oriented in the preferred, normal half-chair ring A conformation.     

Procedure for isolating the endophyte from tall fescue and screening isolates for ergot alkaloids

A procedure was developed to isolate and determine ergot alkaloid production by Acremonium coenophialum, the endophytic fungus of tall fescue. The procedure established that macerated leaf sheath or pith from inflorescence stem placed either in a liquid medium or on a corn meal-malt extract agar medium produced isolated mycelium and characteristic conidia within a 3- to 3.5-week period. Once isolated, each fungus was placed in another liquid medium, M104T, where competent strains produced total ergot alkaloids ranging from 38 to 797 mg/liter. Several isolates were negative for ergot alkaloid synthesis. The production of ergot alkaloids by individual isolates was unstable; isolates rapidly degenerated in their ability to produce ergot alkaloids during subculture. However, the procedure as presented allows the assessment of an isolate for ergot alkaloid synthesis during its initial isolation.     

Procedure for isolating the endophyte from tall fescue and screening isolates for ergot alkaloids.

A procedure was developed to isolate and determine ergot alkaloid production by Acremonium coenophialum, the endophytic fungus of tall fescue. The procedure established that macerated leaf sheath or pith from inflorescence stem placed either in a liquid medium or on a corn meal-malt extract agar medium produced isolated mycelium and characteristic conidia within a 3- to 3.5-week period. Once isolated, each fungus was placed in another liquid medium, M104T, where competent strains produced total ergot alkaloids ranging from 38 to 797 mg/liter. Several isolates were negative for ergot alkaloid synthesis. The production of ergot alkaloids by individual isolates was unstable; isolates rapidly degenerated in their ability to produce ergot alkaloids during subculture. However, the procedure as presented allows the assessment of an isolate for ergot alkaloid synthesis during its initial isolation.     

Biotechnological grounds for the regulation of the component composition of cephalosporins in cultures of Acremonium chrysogenum

The dynamics of cephalosporine C and desacetylcephalosporine C alterations in cultures of Acremonium chrysogenum 309A under different conditions was studied. It was shown expedient to determine the fermantation period by the data of HPLC. Genetic predisposition of the strain used to production of both cephalosporine C and desacetylcephalosporine C is discussed. With changing the cultivation procedure, medium active acidity and cultivation time it is possible to change the biosynthesis pathway.     

Assessment of the metabolic activity of Acremonium chrysogenum using Acridine Orange

A method is described for the assessment of the metabolic activity of the filamentous fungus Acremonium chrysogenum using the fluorescent dye Acridine Orange. Changes in metabolic activity are indicated by a reversible red-green shift in the colour of the dye, quantifiable by image analysis. A. chrysogenum mycelia exhibited an overwhelmingly green colour in circumstances leading to high substrate uptake, while under carbon starvation they appeared orange-red. It is believed these colour changes reflected changes in internal pH. The method provides a visual tool for the investigation of the metabolic behaviour of filamentous fungi.     

A case of colonization of a prosthetic mitral valve by Acremonium strictum

A case of colonization of a prosthetic mitral valve in a 73-year-old Spanish male by the fungus Acremonium strictum W. Gams is described. The valve was replaced due to paravalvular leak and severe insufficiency and the patient died of multiorgan failure. The identity of the fungus was determined by morphological studies and it was confirmed by the analysis of the ITS region sequence analysis. Molecular studies seem to demonstrate that A. strictum is a species complex. The case emphasizes the potential high risk of fungal infection for patients with prosthetic valves.     

PCR assay based on a microsatellite-containing locus for detection and quantification of Epichloë endophytes in grass tissue.

A PCR assay which allows detection and quantification of Epichloë endophytes in tissues of the grass Bromus erectus is described. PCR with specific primers flanking a microsatellite-containing locus (MS primers) amplified fragments 300 to 400 bp in length from as little as 1.0 pg of fungal genomic DNA in 100 ng of DNA from infected plant material. When annealing temperatures were optimized, all Epichloë and Acremonium strains tested, representing many of the known taxonomic groups, yielded an amplification product, indicating that the MS primers may be useful for in planta detection of a variety of related species, including agronomically important Acremonium coenophialum and Acremonium lolii. No fragments were generated from DNA isolates from uninfected plant material or from unrelated fungi isolated from B. erectus. For diagnostic applications, a B. erectus-specific primer pair was designed for use in multiplex PCR to allow simultaneous amplification of plant and fungal DNA sequences, providing an internal control for PCR failure caused by inhibitory plant compounds present in DNA extracts. For quantitative applications, a heterologous control template in primer binding sites complementary to the MS primers was constructed for use in competitive PCR, allowing direct quantification of Epichloë in plant DNA extracts. The fungal DNA present in infected leaves of B. erectus between 1 and 20 pg per 100 ng of leaf DNA, but the amounts of fungal DNA present in the sheath and blade of a given leaf were correlated, indicating that the degree of infection varied between plant individuals but that leaves were colonized in a uniform way.     

Expression of a cephalosporin C esterase gene in <Emphasis Type="Italic">Acremonium chrysogenum</Emphasis> for the direct production of deacetylcephalosporin C

A recombinant fungal microorganism capable of producing deacetylcephalosporin C was constructed by transforming a cephalosporin C esterase gene from Rhodosporidium toruloides into Acremonium chrysogenum. The cephalosporin C esterase gene can be expressed from its endogenous R. toruloides promoter or from the Aspergillus nidulans trpC promoter under standard Acremonium chrysogenum fermentation conditions. The expression of an active cephalosporin C esterase enzyme in A. chrysogenum results in the conversion of cephalosporin C to deacetylcephalosporin C in vivo, a novel fermentation process for the production of deacetylcephalosporin C. The stability of deacetylcephalosporin C in the fermentation broth results in a 40% increase in the cephalosporin nucleus.     

头孢菌素C生物合成调控研究进展北大核心CSCD

头孢菌素C由丝状真菌顶头孢霉产生,属于β-内酰胺类抗生素。其经改造后的7-氨基头孢烷酸是头孢类抗生素的重要中间体。头孢类抗生素在国内外抗生素市场中占有巨大的份额,是临床上的主要抗感染药物。随着分子生物学的发展,头孢菌素C的生物合成途径已基本阐明。为提高头孢菌素C的产量和降低生产成本,越来越多的研究者开始关注其较为精细、复杂的调控机制。本文重点对头孢菌素C生物合成及其调控机制的最新进展进行了简述,希望为今后头孢菌素C生产菌株的菌种改造和传统产业的升级换代提供一定的借鉴。     

Characterisation of the extracellular polysaccharides produced by isolates of the fungus Acremonium

Solid state (13)C NMR studies of the extracellular glucans from the fungi Acremonium persicinum C38 (QM107a) and Acremonium sp. strain C106 indicated a backbone of (1-->3)-beta-linked glucosyl residues with single (1-->6)-beta-linked glucosyl side branches for both glucans. Analyses of enzymatic digestion products suggested that the average branching frequency for the A. persicinum glucan (66.7% branched) was much higher than that of the Acremonium sp. strain C106 glucan (28.6% branched). The solid state (13)C NMR spectra also indicated that both glucans are amorphous polymers with no crystalline regions, and the individual chains are probably arranged as triple helices.     

顶头孢霉遗传育种研究进展

顶头孢霉是一类重要的工业微生物,其发酵产物头孢菌素C可用来生产7-ACA,而后者是临床常用抗感染药物头孢类抗生素的重要中间体。头孢菌素C的发酵水平决定了其下游头孢类抗生素的生产水平、产品质量及价格,因此对顶头孢霉的菌种选育工作显得尤其迫切。随着分子生物学的发展,基因工程分子改造在遗传育种领域发挥着越来越重要的作用。文章综述了对头孢菌素C的生物合成以及调控的研究进展,并将国内外对顶头孢霉进行遗传育种的结果进行了归纳总结,提出了可以从提高头孢菌素C发酵水平、延伸代谢途径等不同方面对头孢菌素C生物合成及调控基因,包括外源基因的导入和表达进行改造优化,并对进一步的研究目标进行了展望,认为可以结合比较蛋白质组和基因组改组使遗传育种所获得的工程菌尽快进入产业化。     

Fungal Pathogen of Cladophora glomerata (Chlorophyta)

A strain of Acremonium kiliense (Fungi Imperfecti) produced a water-soluble, dialyzable, heat-stable agent that rendered Cladophora glomerata (Chlorophyta) chlorotic and inhibited its growth.     

Study of the optimal conditions of preservation of cephalosporin C-producing fungus Acremonium chrysogenum

The influence of various preservation conditions of viability and antibiotic activity of Acremonium chrysogenum 281A and 305A strains producing cephalosporin C was studied. Cryogenization of the culture in the form of suspension of the vegetative mycelium in 20 per cent glycerol solution at a rate of 1 degree/min showed to be advantageous over lyophilization and L-drying. Cryogenization under such conditions provided rather high viability of the culture and preservation of its initial antibiotic activity for the period of its storage for at least 1.5 years in liquid nitrogen at a temperature of -196 degrees C.     

Relationships among non-Acremonium sp. fungal endophytes in five grass species.

Many cool-season grasses (subfamily Pooideae) possess maternally transmitted fungal symbionts which cause no known pathology and often enhance the ecological fitness and biochemical capabilities of the grass hosts. The most commonly described endophytes are the Acremonium section Albo-lanosa spp. (Acremonium endophytes), which are conidial anamorphs (strictly asexual forms) of Epichloë typhina. Other endophytes which have been noted are a Gliocladium-like fungus in perennial ryegrass (Lolium perenne L.) and a Phialophora-like fungus in tall fescue (Festuca arundinacea Schreb.). Here, we report the identification of additional non-Acremonium sp. endophytes (herein designated p-endophytes) in three more grass species: Festuca gigantea, Festuca arizonica, and Festuca pratensis. In each grass species, the p-endophyte was cosymbiotic with an Acremonium endophyte. Serological analysis and sequence determinations of variable portions of their rRNA genes indicated that the two previously identified non-Acremonium endophytes are closely related to each other and to the newly identified p-endophytes. Therefore, the p-endophytes represent a second group of widely distributed grass symbionts.     

黑刺粉虱8种虫生真菌培养性状及其侵染率

蚧侧链孢（Pleurodesmospora coccora）、韦伯虫座孢（Aegerita webberi）、枝孢霉（Cladosporium sp.）、顶孢霉（Acremonium sp.）、拟青霉（Paecilomyces sp.）、米曲霉（Aspergillus oryzae）、被毛孢（Hirsutella sp.）和祁门4号（学名？）是茶园黑刺粉虱（Aleurocanthus spiniferus）主要的虫生真菌。它们在马铃薯培养基、萨氏培养基和察氏培养基上生长状况较好,但韦伯虫座孢不能产生有性世代。中描述了它们在察氏平板培养基上25℃恒温培养3星期的菌落特征围 伯虫座孢是皖南山区茶园中的优势种,“霉雨”季节侵染率达90％以上。蚧侧链孢、顶孢霉和枝孢霉是皖南丘陵地区茶园中优势种,流行盛期的联合侵染率为0.4%-11%。     

Production, Characterization, and Use of Monoclonal Antibodies Against Acremonium coenophialum

Monoclonal antibodies were produced using intact mycelium of the fungus Acremonium coenopbialum as the immunogen. During the initial stages of characterization, the antibodies were found to react to A. coenophialum, A. loliae, Epichloe typhina , and also to cross-react with some other fungi normally associated with tall fescue. Careful selection of the ELISA system in which the antibodies were used eliminated reactions to all but the two Acremonium spp. and E. typhina. Asa result it was possible to detect Acremonium spp. (presumably A. coenophialum ) sensitively and unambiguously in leaf tissue of tall fescue.     

Azolla filiculoides Nitrogenase Activity Decrease Induced by Inoculation with Chlamydomonas sp

Experiments were conducted to determine the influence of Chlamydomonas sp. on nitrogen fixation (C(2)H(2) --> C(2)H(4)) in Azolla filiculoides and on the nitrogen fixation and growth of free-living Anabaena azollae 2B organisms. Inoculation of azolla medium with Chlamydomonas sp. was associated with decreased nitrogenase activity in A. filiculoides and with increases in the density of a fungal population identified as Acremonium sp. Subsequent inoculation of azolla medium with this fungus was also accompanied by a significant decrease in nitrogenase activity of A. filiculoides. However, the extent of depression of nitrogenase activity was significantly higher when azolla medium was inoculated with Chlamydomonas sp. than when it was inoculated with Acremonium sp. Inoculation of nitrogen-free Stanier medium with either Acremonium sp. or Chlamydomonas sp. did not adversely affect the growth or nitrogenase activity of free-living A. azollae. Decreased nitrogenase activity in A. filiculoides is apparently related to the adverse influence of the green alga and the fungus on the macrosymbiont. The mechanisms that might be involved are discussed.     

枝顶孢霉对毒光盖伞子实体形成的影响

报道枝顶孢霉Acremonium strictum对毒光盖伞Psilocybe venenata子实体形成的影响。拮抗试验结果表明它们之间存在着较弱的干扰竞争作用;人工培养时,毒光盖伞混合菌株产生大型、小型和畸形3种类型的子实体,推测毒光盖伞畸形子实体的形成可能是枝顶孢霉寄生的结果,为毒光盖伞的人工栽培和进一步合理利用、野生蕈菌的驯化提供了理论依据。     

中国的枝顶孢属

本文对中国的枝顶孢属丝孢菌进行了调查,其中包括3个新记录种,并给出了全部13个种的检索表。