Isolation of feeding deterrents against argentine stem weevil from ryegrass infected with the endophyteAcremonium loliae

Infection of ryegrass (Lolium perenne) by an endophytic fungus (Acremonium loliae) confers resistance against the Argentine stem weevil (Listronotus bonariensis). Extracts from ryegrass clones, infected and uninfected withA. loliae, were compared in a feeding choice bioassay, and several fractions were identified which affected stem weevil feeding behavior. One stem weevil feeding deterrent, peramine C₁₂H₁₇N₅O, has been isolated from infected ryegrass and partially characterized as a basic indole derivative. Extracts from culturedA. loliae had no effect on stem weevil feeding behavior nor was peramine detected in the fungal cultures examined. Peramine and the other active substances are hydrophilic in contrast to the lipophilic properties reported for the neurotoxic lolitrems also isolated from ryegrass infected withA. loliae and associated with causing ryegrass staggers disorder in livestock. It is suggested that ryegrass staggers and stem weevil feeding deterrency may arise by different biochemical mechanisms.     

Ergovaline and peramine production in endophyte-infected tall fescue: Independent regulation and effects of plant and endophyte genotype

Peramine and ergovaline have ecological and economical significance because they possess insect and/or mammalian toxicity properties. The relationship between these endophytically derived alkaloids in tall fescue (Festuca arundinacea Schreb.) is unknown. Seasonal and plant tissue effects on the concentration of peramine and ergovaline was investigated in field and greenhouse experiments. The relationship between the alkaloids and the regulatory effects of endophyte and plant on their content were investigated among progeny of reciprocal crosses between high- and low-ergovaline and peramine plant-endophyte combinations. Variation in peramine concentration ranged from 750 to 1742 µg/kg in greenhouse-grown plants, and there was no seasonal trend in peramine content. There was a correlation (r=0.69) between peramine and ergovaline content among leaf tissues of field-grown plants, but there was no correlation between the alkaloids in the culm (r=0.20) or panicle (r=0.17) tissues. Mean leaf ergovaline concentration of progeny derived from the low-ergovaline parent (163 µg/kg) was less than the midparent value (228 µg/kg), but mean of progeny from the high-ergovaline parent was not different from the mid-parent value. Ranges within each progeny set were approximately double their mean. Mean leaf peramine concentrations of the progeny sets were similar to the mid-parent value (3354 µg/kg) but ranges were from 1716 to 8753 µg/kg. There was no correlation between leaf peramine and ergovaline (r=0.01). These data suggest that endophytically produced alkaloids are independently regulated and are controlled by both plant and endophyte genotype.     

Indoor mold levels and current asthma among school‐aged children in Saskatchewan,Canada

Current knowledge regarding the association between indoor mold exposures and asthma is still limited. The objective of this case–control study was to investigate the relationship between objectively measured indoor mold levels and current asthma among school‐aged children. Parents completed a questionnaire survey of health history and home environmental conditions. Asthma cases had a history of doctor‐diagnosed asthma or current wheeze without a cold in the past 12 months. Controls were age‐ and sex‐matched to cases. Vacuumed dust samples were collected from the child's indoor play area and mattress. Samples were assessed for mold levels and quantified in colony‐forming units (CFU). Sensitization to mold allergens was also determined by skin testing. Being a case was associated with family history of asthma, pet ownership, and mold allergy. Mold levels (CFU/m²) in the dust samples of children's mattress and play area floors were moderately correlated (r = 0.56; P < 0.05). High mold levels (≥30 000 CFU/m²) in dust samples from play [adjusted odds ratio (aOR) = 2.6; 95% CI: 1.03–6.43] and mattress (aOR) = 3.0; 95% CI: 1.11–8.00) areas were significantly associated with current asthma. In this study high levels of mold are a risk factor for asthma in children.     

Domestic exposure to fungal allergenic particles determined by halogen immunoassay using subject's serum versus particles carrying three non‐fungal allergens determined by allergen‐specific HIA

Studies that estimate indoor aeroallergen exposure typically measure a pre‐selected limited range of allergens. In this study, inhalable aeroallergen particles were quantified using the halogen immunoassay (HIA) to determine the contribution of fungal and non‐fungal aeroallergens to total allergen exposure. Bioaerosols from 39 homes of fungal‐allergic subjects were sampled using inhalable fraction samplers and immunostained by HIA using resident subject's immunoglobulin E (IgE) to detect allergen‐laden particles. Fungal aerosols as well as particles carrying mite, cat, and cockroach allergens were identified and enumerated by HIA. Reservoir dust‐mite (Der p 1), cat (Fel d 1), and cockroach (Bla g 1) allergen concentrations were quantified by ELISA. Fungal particles that bound subject's IgE in the HIA were 1.7 (bedroom)‐ and 1.4 (living room)‐fold more concentrated than Der p 1, Fel d 1, and Bla g 1 allergen particles combined. Predominant fungal conidia that bound IgE were derived from common environmental genera including Cladosporium and other fungi that produce amerospores. Airborne mite, cat, and cockroach allergen particle counts were not associated with reservoir concentrations determined by ELISA. This study demonstrates that inhalable fungal aerosols are the predominant aeroallergen sources in Sydney homes and should be considered in future exposure assessments.     

Inhibitory of multiple antifungal components produced by Lactobacillus plantarum K35 on growth,aflatoxin production and ultrastructure alterations of Aspergillus flavus and Aspergillus parasiticus

The antifungal activity of Lactobacillus plantarum K35 isolated from traditional Thai fermented rice noodle was evaluated against the growth and aflatoxin production of Aspergillus flavus TISTR304 and Aspergillus parasiticus TISTR3276. Multiple antifungal compounds secreted by L. plantarum were firstly analyzed using GC–MS in parallel to the ultrastructure alteration of the treated fungi. L. plantarum K35 supernatant caused significant reduction of the fungal growth and aflatoxin production. The antifungal activity was pH-dependent and favorable to acidic conditions whereas the catalase treatment had no influence indicating no involvement of hydrogen peroxide in the inhibition. Antifungal substances were resistant to sterilization and proteolytic enzymes including trypsin and proteinase K. The major components, apart from lactic acid, were 2-butyl-4-hexyloctahydro-1H-indene (19.55%), oleic acid (10.52%) and palmitic acid (7.27%). Other minor antifungal compounds included linoleic acid (2.11%), 2,4-di-tert-butylphenol (1.84%), stearic acid (1.55%), 3-phenyllactic acid (1.42%) and pyroglutamic acid (1.07%). The morphological changes of the fungi exposed to the supernatant were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). L. plantarum K35 supernatant caused severe damage to the cell wall and cytoplasmic membrane leading to a massive loss of cytoplasmic content, the formation of membrane-bound vesicles, and complete destruction of membranous organelles including mitochondria and nucleus.     

植物内生菌及其次生代谢活性物质多样性的研究进展

植物内生菌是一种新型的微生物资源,具有重大的研究意义和潜在的应用价值,近年来已成为微生物资源研究的热点之一.对植物内生菌及其次生代谢活性物质多样性的研究进展进行了综述.     

BUTYL ACETATE AND YEASTS INTERACT IN ADHESION AND GERMINATION OF Botrytis cinerea CONIDIA IN VITRO AND IN FUNGAL DECAY OF GOLDEN DELICIOUS APPLE

Butyl acetate is a volatile aroma and flavor compound in apple. Conidia of three strains of Botrytis cinerea, a fungus that causes decay of apple fruit in postharvest storage, had greater adhesion to and greater germination on polycarbonate membrane filters on water inside sealed 500 cc glass jars that were injected with 4 l butyl acetate than conidia not so exposed. Conidial germination was highly correlated with conidial adhesion. The yeasts Sporobolomyces roseus and Cryptococcus laurentii, but not Saccharomyces cerevisiae, reduced the adhesion and germination promoting effect of butyl acetate. Conidia did not readily utilize butyl acetate as a food source, as shown by lack of tetrazolium violet reduction, whereas S. roseus and C. laurentii, but not S. cerevisiae did. Butyl acetate added to suspensions of conidia increased the electrical conductivity of the suspensions and increased the loss of ¹⁴C from ¹⁴C-labeled conidia compared to conidia unexposed to butyl acetate. Uptake of [¹⁴C]glucose by conidia was not increased by butyl acetate. Wounds of Golden Delicious apples inoculated with conidia (strain F-J-4) in a dilute solution of butyl acetate had greater decay than unexposed wounds. S. roseus and C. laurentii, but not S. cerevisiae, added with the conidia decreased the incidence or size of decay. Results indicated that butyl acetate increased conidial adhesion, stimulating conidial germination, and some yeasts can reduce this effect.     

Gene expression profiling in streptozotocin-induced diabetic rat liver in response to fungal polysaccharide treatment

We established the gene expression profiling in streptozotocin (STZ)-induced diabetic rat liver in response to hypoglycemic fungal polysaccharides (EPS), using oligonucleotide microarray analysis. Differentially regulated genes showing higher fold change than 2 were identified and categorized through hierarchical clustering analysis. Among the 835 genes analyzed, 244 were up-regulated, while 321 were down-regulated after diabetes induction. Interestingly, many gene expressions altered after STZ-treatment mimicked a normal rat liver by EPS therapy. Most of these genes included genes involving cell structure and motility, immunity and defense, lipid, fatty acid and steroid metabolism, protein metabolism and modification, and signal transduction. More importantly, we found a total of 36 genes that showed significant fold changes in their expression that have not previously been examined in the context of diabetes. To validate the microarray results, we further confirmed the gene expression patterns by RT-PCR using four genes of interest (carboxylesterase 2, stearoyl-coenzyme A desaturase 1, insulin-like growth factor 1, and insulin-like growth factor binding protein 2). Taken together, EPS acted as a potent regulator of gene expression for a wide variety of genes in diabetic rat liver.     

Chemotaxis Disruption in Pratylenchus Scribneri by Tall Fescue Root Extracts and Alkaloids

Tall fescue (Festuca arundinacea) forms a symbiotic relationship with the clavicipitalean fungal endophyte Neotyphodium coenophialum. Endophyte-infected grass is tolerant to nematode, but the factors responsible are unknown. One objective of this work was to determine if root extracts of tall fescue effected chemoreceptor activity of Pratylenchus scribneri by using an in vitro chemoreception bioassay. Another objective was to determine if specific ergot alkaloids (ergovaline, ergotamine, a-ergocryptine, ergonovine), and loline alkaloids, all produced by the fungal endophyte, altered chemotaxis with this bioassay. Methanolic extract from roots altered chemotaxis activities in this nematode but only from roots of plants cultured 45 ≥ d, which repelled nematodes. Extracts prepared from noninfected grasses were attractants. This assay indicated that the alkaloids were either repellents or attractants. N-formylloline was an attractant at concentrations of 20 μg/ml and lower, while at higher concentrations it was a repellent. Ergovaline, the major ergot alkaloid produced by the endophyte, was repellent at both high and low concentrations and caused complete death of the nematodes.