Fungal colonization of synthetic substrates for use in space craft

Summary Materials being used or considered for use in space flight were examined for their susceptibility to fungal colonization. The materials included soft goods (clothing) and insulation and fabrication products such as Velcro^® attachments and elastic cord binders. Materials were exposed for at least 28 days in a highhumidity chamber colonized with over 50 species of fungi, including those species recommended for determining recalcitrance of materials to fungal biodegradation. At least nine of 25 products demonstrated extensive microscopic colonization by fungi, mostly byAcremonium obclavatum. Challenge procedures that rely on observations with the unaided eye, or 40×magnification of growth by a restricted number of fungal species with a cellulosic substrate as a positive control, are insufficient for determining the resistance of synthetic substrates to fungal colonization.     

寄生双孢蘑菇的蛛网丝枝霉中国变种

1980—1981年自北京、福州、三明、泉州、漳州、厦门和昆明等地双孢蘑菇[Agaricusbisporus(Lange)Sing]病菇上和不出蘑菇的培养料内,分离到一种真菌。在 PDA 培养基上的培养性状,肉眼观察与蘑菇轮枝菌(Verticillium psalliotae Tresch.)极难区别。根据其分生孢子和瓶梗的形态及其在匐匍气丝上的排列情况等,鉴定为珠网丝枝霉。因具短棒状原瓶梗,认为是一新变种,定名为珠网丝枝霉中国变种 Aphanocladium aranearum(Petch)W.Gams var.sinense J.D.Chen var.nov.它是双孢蘑菇的重要害菌之一。这个“属”和“种”都是我国的新记录。在猴头(Hericium erinaceus)、香菇(Lentinus edo-des),糙皮侧耳(Pleurotus ostreatus)上和水稻土中也有少量分布。     

Diversity and origins of endophytic fungal symbionts of the North American grass Festuca arizonica

Summary Acremonium spp. endophytes are mutualistic fungal symbionts of many C3 grasses. They are anamorphs of Epichloë typhina (Clavicipitaceae) that have become strictly seedborne, heritable components of symbiotic units (symbiota). In order to test the possibility that endophytes may contribute to the genetic diversity of symbiota, a survey was conducted of plants from nine populations of Festuca arizonica in the southern Rocky Mountains. Sequence analysis of rRNA gene segments distinguished three Acremonium endophyte types. Parsimony analysis indicated at least two distinct evolutionary origins of the Acremonium endophytes from E. typhina. Either or both of these evolutionary lineages may have involved cospeciation with the host.     

Susceptibility of tall fescue to Rhizoctonia zeae infection as affected by endophyte symbiosis

Endophytic fungi belonging to the genus Neotyphodium often form symbiotic associations with grasses. The host plants usually benefit from the association with an endophyte. Presence of the symbiont may increase host resistance to infection by some pathogens. However, the exact mechanism of the lower susceptibility of endophyte‐infected plants to diseases is still unclear. Growth chamber trials were conducted to determine whether (a) tall fescue plants infected with the endophyte Neotyphodium coenophialum (E+) are more resistant to sheath and leaf spot disease caused by Rhizoctonia zeae than endophyte‐free (E?) plants, and (b) R. zeae growth inhibition is associated with endophyte presence. Tall fescue genotypes, each symbiotic with a genetically different native endophyte strain, were inoculated with isolates of R. zeae. The tillers infection by R. zeae, density of endophyte hyphae and content of total phenolic compounds in tillers were studied. Antifungal activity of the N. coenophialum towards R. zeae, Rhizoctonia solani, Bipolaris sorokiniana and Curvularia lunata was also investigated in dual‐culture assays. For Tf3, Tf4, TfA2 and TfA9 tall fescue genotypes, the E+ plants had reduced R. zeae infection. In the Tf9 and Tf8085 genotypes, R. zeae infection was similar for both E+ and E? plants. The strongest effect was observed for the Tf4 endophyte. A strongly positive correlation (r = 0.94) occurred between endophyte hyphal density and disease index across all tall fescue genotypes. Dual‐culture assays showed no inhibitory interaction between the seven endophyte strains and the R. zeae isolates; however, some endophytes inhibited R. solani, B. sorokiniana and C. lunata. Endophyte presence increased the production of phenolic compounds by the host grasses. The level of phenolics also differed significantly depending on the time of analysis after inoculation of plants by R. zeae. The results indicate that N. coenophialum can suppress disease severity caused by R. zeae infection. The mechanism of higher resistance of E+ plants is likely not based on direct inhibition such as antibiosis or competition. Thus, the induction of specific mechanisms in the host plant, for example, production of phenolic compounds, seems to be the main way of providing resistance to the grass by the endophyte.     

Inactivation of Bacteriophage ϕX174 by d-Fructose 6-Phosphate

The inactivation of bacteriophage ?X174 by d-fructose 6-phosphate was investigated. This inactivation was inhibited by EDTA or reducing agents, and stimulated by Cu²⁺ but other metal ions could not be substituted for Cu²⁺. The reaction was also inhibited by superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6) and various free radical scavengers.

No detectable changes were observed in adsorption capacity of phage and in the conformation of the virion. The viral DNA in the virion was, however, found to be cleaved. This strand scission was also enhanced by Cu²⁺ and protected by catalase. Similar results were obtained when ?X174 DNA was directly treated with d-fructose 6-phosphate.

It is concluded that the inactivation of ?X174 is due to DNA strand scission in the virion by the free radical of d-fructose 6-phosphate or oxygen radicals generated during autoxidation of d-fructose 6-phosphate.     

Heterologous Protein Production in Acremonium chrysogenum: Expression of Bacterial Cephalosporin C Acylase and Human Thrombomodulin Genes

We have developed an efficient expression system for foreign genes in Acremonium chrysogenum. After inserting the foreign gene between the phosphoglycerate kinase (PGK) promoter and a terminator derived from A. chrysogenum, multiple copies of this expression unit are tandemly ligated into cosmids and the resultant cosmids are introduced into A. chrysogenum.

We expressed Pseudomonas cephalosporin C acylase and a human thrombomodulin mutant protein containing the fourth, fifth, and sixth epidermal growth factor (EGF)-like structures (E456). The acylase activity in the transformants obtained using our system was several times higher than that in the transformants without the use of the system. The acylase proteins expressed had enzymatic and immunochemical properties identical to those of authentic acylase. The transformants with the expression plasmid for E456 secreted biologically active E456 protein into the culture medium. The amino terminal sequence of the purified E456 was identical to that of recombinant E456 obtained using mammalian cells.     

Microbial hydroxylation of progesterone with Acremonium strictum

Microbial hydroxylation of progesterone occurred in the culture of Acremonium strictum PTCC 5282 to produce two hydroxylated pregnene-like steroids. The metabolites were purified and characterized using spectroscopic methods and identified as 15alpha-hydroxyprogesterone and 15alpha-hydroxydeoxycorticosterone.     

Influence of phosphorus on the growth and ergot alkaloid content of Neotyphodium coenophialum-infected tall fescue (Festuca arundinacea Schreb.)

Tall fescue (Festuca arundinacea Schreb.) plants infected by the fungal endophyte Neotyphodium coenophialum (Morgan-Jones & Gams) (Glenn et al., 1996) often perform better than noninfected plants, especially in marginal resource environments. There is a lack of information about endophyte related effects on the rhizosphere of grasses. In a greenhouse experiment, four endophyte-infected (E+) tall fescue clones (DN2, DN4, DN7, DN11) and their endophyte-free (E–) forms were grown in limed (pH 6.3) Porter soil (low fertility, acidic, high aluminum and low phosphorus content, coarse-loamy mixed mesic Umbric Dystrochrept) at three soil P levels (17, 50, and 96 mg P kg^-1 soil) for five months. Excluding the genotype effect, endophyte infection significantly increased cumulative herbage DM yield by 8% at 17 mg P kg^-1 soil but reduced cumulative herbage DM yield by 12% at 96 mg P kg^-1 soil. With increased P availability in the soil, shoot and root DM, and root/shoot ratio in E+ plants were significantly less when compared to E– plants. Endophyte infection increased specific root length at 17 and 50 mg P kg^-1soil. At soil P level of 17 mg P kg^-1soil, E+ plants had significantly higher P concentrations both in roots and shoots. Similar relationships were found for Mg and Ca. E+ plants had significantly higher Zn, Fe, and Al concentration in roots, and lower Mn and Al concentration in shoots when compared to E– plants. Ergot alkaloid concentration and content in shoot of E+ plants increased with increasing P availability in the soil from 17 to 50 mg P kg^-1 but declined again at 96 mg P kg^-1 soil. Ergot alkaloid accumulation in roots increased linearly with P availability in the soil. Results suggest that endophyte infection affects uptake of phosphorus and other mineral nutrients and may benefit tall fescue grown on P-deficient soils. Phosphorus seems also to be involved in ergot alkaloid accumulation in endophyte-infected tall fescue.     

Efficient production of cellulase in the culture of Acremonium cellulolyticus using untreated waste paper sludge

Cellulase was produced by Acremonium cellulolyticus using untreated waste paper sludge (PS) as the carbon source. The clay present in PS did not show any inhibitory effect on cellulase production but did alter the pH during fermentation. On the flask scale, the maleate buffer concentration and pH were key factors that affected the efficiency of cellulase production from PS cellulose. Optimum cellulase production in a 3-L fermentor of working volume 1.5 L was achieved by controlling the pH value at 6.0 using 2 M NaOH and 2 M maleic acid, and the productivity reached 8.18 FPU/mL. When 40.89 g/L PS cellulose, 2.2 g/L (NH(4) )(2) SO(4) , and 4.4 g/L urea were added to a 48-h culture, the cellulase activity was 9.31 FPU/mL at the flask scale and 10.96 FPU/mL in the 3-L fermentor. These values are ～80% of those obtained when pure cellulose is used as the carbon source. The method developed here presents a new route for the utilization of PS.