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In this paper, we demonstrate the ability of Arabidopsis thaliana to detect different mixtures of volatile organic compounds (VOCs) emitted by the common indoor fungus, Aspergillus versicolor, and demonstrate the potential usage of the plant as a bioindicator to monitor fungal VOCs in indoor air. We evaluated the volatile production of Aspergillus versicolor strains SRRC 108 (NRRL 3449) and SRRC 2559 (ATCC 32662) grown on nutrient rich fungal medium, and grown under conditions to mimic the substrate encountered in the built environment where fungi would typically grow indoors (moist wallboard and ceiling tiles). Using headspace solid phase microextraction/gas chromatography-mass spectrometry, we analyzed VOC profiles of the two strains. The most abundant compound produced by both strains on all three media was 1-octen-3-ol. Strain SRRC 2559 made several terpenes not detected from strain SRRC 108. Using a split-plate bioassay, we grew Arabidopsis thaliana in a shared atmosphere with VOCs from the two strains of Aspergillus versicolor grown on yeast extract sucrose medium. The VOCs emitted by SRRC 2559 had an adverse impact on seed germination and plant growth. Chemical standards of individual VOCs from the Aspergillus versicolor mixture (2-methyl-1-butanol, 3-methyl-1-butanol, 1-octen-3-ol, limonene, and β-farnesene), and β-caryophyllene were tested one by one in seed germination and vegetative plant growth assays. The most inhibitory compound to both seed germination and plant growth was 1-octen-3-ol. Our data suggest that Arabidopsis is a useful model for monitoring indoor air quality as it is sensitive to naturally emitted fungal volatile mixtures as well as to chemical standards of individual compounds, and it exhibits relatively quick concentration- and duration-dependent responses.

The current trends to (1) investigate sugarcane leaves as a sustainable biomass feedstock for the production of biofuels and bioproducts and (2) delivery of more leaves to factories for processing with stalks, have made information on how it deteriorates on storage during dry and wet environmental conditions more important. Brown, dry (BL) and green leaves (GL) were stripped from hand-cut whole-stalks from Louisiana (LA) sugarcane variety LCP 85-384 in two consecutive years. In the first year, BL and GL were separately allowed to deteriorate in a simulated field canopy shade study. One set was watered across 14 days to simulate rain, and the other set was not watered. This 14 day period was repeated three times across the LA harvest season (October to December) to cover varying environmental conditions: 17–31 October (21 °C; 71% mean humidity), 15–29 November (22 °C; 71%), and 13–27 December (15.5 °C; 78%). In the second year, the simulated study was repeated but over a prolonged 28 day period (17 November–15 December) and fresh and deteriorated leaves were also collected from a grower's LCP 85-384 field and analyzed for comparison. Generally, the worst deterioration for both BL and GL occurred in the watered samples, and when the humidity was highest. On deterioration, more soluble impurities were extracted from GL than BL. Only prolonged deterioration of BL caused a reduction of fiber biomass, and the fiber content of GL usually increased on deterioration because of loss of moisture. The nutrient sucrose was often, but not always, preferentially utilized over glucose and fructose. Results from the grower's field were more extreme than from the simulated experiments. Leaf moisture was higher in the deteriorated than non-deteriorated field BL because its shredded state created more surface area to absorb water; this also allowed for more Leuconostoc mesenteroides bacteria to grow and form dextran and mannitol. Overall, the total microbe populations were highest on watered GL because of higher available nutrients.

Summary The domestication of animals, plants, and microbes fundamentally transformed the lifestyle and demography of the human species [1]. Although the genetic and functional underpinnings of animal and plant domestication are well understood, little is known about microbe domestication [2–6]. Here, we systematically examined genome-wide sequence and functional variation between the domesticated fungus Aspergillus oryzae , whose saccharification abilities humans have harnessed for thousands of years to produce sake, soy sauce, and miso from starch-rich grains, and its wild relative A. flavus , a potentially toxigenic plant and animal pathogen [7]. We discovered dramatic changes in the sequence variation and abundance profiles of genes and wholesale primary and secondary metabolic pathways between domesticated and wild relative isolates during growth on rice. Our data suggest that, through selection by humans, an atoxigenic lineage of A. flavus gradually evolved into a "cell factory" for enzymes and metabolites involved in the saccharification process. These results suggest that whereas animal and plant domestication was largely driven by Neolithic "genetic tinkering" of developmental pathways, microbe domestication was driven by extensive remodeling of metabolism.

article i nfo The Aspergillus niger aggregate within the A. section Nigri is a group of black-spored aspergilli of great agro- economic importance whose well defined taxonomy has been elusive. Rep-PCR has become a rapid and cost- effective method for genotyping fungi and bacteria. In the present study, we evaluated the discriminatory power of a semi-automated rep-PCR barcoding system to distinguish morphotypic species and compare the results with the data obtained from ITS and partial calmodulin regions. For this purpose, 20 morphotyped black-spored Aspergillus species were used to create the A. section Nigri library in this barcoding system that served to identify 34 field isolates. A pair-wise similarity matrix was calculated using the cone-based Pearson correlation method and the dendrogram was generated by the unweighted pair group method with arithmetic mean (UPGMA), illustrating four different clustered groups: the uniseriate cluster (I), the As- pergillus carbonarius cluster (II), and. the two A. niger aggregate clusters (named III.A and III.B). Rep-PCR showed higher resolution than the ITS and the partial calmodulin gene analytical procedures. The data of the 34 unknown field isolates, collected from different locations in the United States, indicated that only 12% of the field isolates were N95% similar to one of the genotypes included in the A. section Nigri library. However, 64% of the field isolates matched genotypes with the reference library (similarity values N90%). Based on these results, this barcoding procedure has the potential for use as a reproducible tool for identifying the black-spored aspergilli. Published by Elsevier B.V.

Aspergillus species are increasingly important human pathogens. It is not known whether toxic metabolites of many of these pathogenic species can act as virulence factors in aspergillosis. We examined isolates of aflatoxin and ochratoxin-producing species for toxin production in ex vivo conditions. Seven of the 21 aflatoxin-producing isolates screened produced aflatoxin at 35 and 37 degrees C on the general medium yeast extract sucrose agar (YES). However, none of them produced toxin at these temperatures on brain heart infusion agar (BHA), a medium that mimics human tissue, or on BHA with modified pH or sugar levels. Six of the 12 ochratoxin-producing isolates examined produced toxin at 35 degrees C on YES. All three isolates of A. alliaceus produced ochratoxin on BHA or modified BHA at 37 degrees C. One strain of A. pseudoelegans produced a minute amount of ochratoxin on modified BHA at 37 degrees C. These data indicate that aflatoxin is an unlikely virulence, factor but that ochratoxin may be a potential virulence factor in aspergillosis.

Production of carcinogenic aflatoxins has been reported from members of Aspergillus section Flavi, Aspergillus section Nidulantes and a newly proposed Aspergillus section Ochraceorosei that consists of Aspergillus ochraceoroseus and A. rambellii. Unlike members of section Flavi, A. ochraceoroseus and A. rambellii have been shown to accumulate both aflatoxin (AF) and the aflatoxin precursor sterigmatocystin (ST). Alhough morphologically distinct from A. nidulans, molecular characterization of A. ochraceoroseus AF/ST genes and physiological characteristics of AF/ST production indicated that A. ochraceoroseus is more closely related to A. nidulans than to A. flavus. Knowing that the A. nidulans ST gene cluster is organized differently from the A. flavus AF gene cluster, we determined the genetic organization of the AF/ST biosynthetic cluster in A. ochraceoroseus. Sequencing of overlapping lambda clones and genomic PCR fragments obtained by gene-walking techniques demonstrated that the A. ochraceoroseus AF/ST gene cluster is organized much like the A. nidulans ST gene cluster except that the region from aflN to aflW is located directly upstream of aflC and in reverse orientation such that aflW represents the distal end and aflY the proximal end of the cluster. The A. ochraceoroseus cluster genes demonstrated 62-76% nucleotide identity to their A. nidulans ST cluster gene homologs. Transformation of an A. nidulans aflR mutant with the A. ochraceoroseus aflR restored ST production in A. nidulans transformants. PCR amplification of A. rambellii genomic DNA demonstrated that the AF/ST gene cluster is organized in the same manner as that of A. ochraceoroseus.

Aflatoxins are carcinogenic mycotoxins formed by a number of fungi in the genus Aspergillus. The major fungi responsible for aflatoxin formation in crop seeds in the field and in storage are Aspergillus flavus and A. parasiticus. This review emphasizes developmental, environmental, biological, and chemical factors that influence aflatoxin formation by A. flavus and A. parasiticus.

As the number of cases of aspergillosis grows, the number of species reported to cause the disease is increasing. Historically, classification and identification of aspergilli was accomplished using morphological characteristics. A number of molecular, immunological and biochemical methods are now available. For the most part, the results of the various approaches concur, yielding similar results in identifying aspergilli, so the 'best' method for identification is the method that best suits the needs of the researcher or clinician. Each identification method has advantages and disadvantages which will be discussed herein. The paper contains a listing of some of the available identification systems from those that address the whole genus to those capable of separating intraspecific strains.

Most aspergilli that produce aflatoxin are members of Aspergillus section Flavi, however isolates of several Aspergillus species not closely related to section Flavi also have been found to produce aflatoxin. Two of the species, Aspergillus ochraceoroseus and an undescribed Aspergillus species SRRC 1468, are morphologically similar to members of Aspergillus section Circumdati. The other species have Emericella teleomorphs (Em. astellata and an undescribed Emericella species SRRC 2520) and are morphologically distinctive in having ascospores with large flanges. All these aflatoxin-producing isolates were from tropical zones near oceans, and none of them grew on artificial media at 37 C. Aflatoxins and sterigmatocystin production were quantified by high-pressure liquid chromatography (HPLC) and confirmed by HPLC-mass spectrometry (LC-MS) detection. Phylogenetic analyses were conducted on these four species using A. parasiticus and Em. nidulans, (which produce aflatoxin and the aflatoxin precursor sterigmatocystin, respectively) for comparison. Two aflatoxin/sterigmatocystin biosynthesis genes and the beta tubulin gene were used in the analyses. Results showed that of the new aflatoxin-producers, Aspergillus SRRC 1468 forms a strongly supported clade with A. ochraceoroseus as does Emericella SRRC 2520 with Em. astellata SRRC 503 and 512.

Based on counts of Aspergillus species reported in over 250 studies of microfungi from soils and litter, chi-square analyses were conducted on species occurrence in five biomes and five latitude ranges to determine variations from expected distributions. There was no overall trend in distribution of the members of the entire genus by biome, however, individual sections of the genus appeared to have distinct distribution patterns. Most members of sections Aspergillus, Nidulantes, Flavipedes and Circumdati occurred at greater than expected frequencies in desert soils. There was no distinct pattern of species occurrence for forest, wetland, or cultivated soils, although members of section Nidulantes were quite rare in cultivated soils. Most species occurred at or below expected frequencies in grassland soils. Members of the genus tended to occur at greater than expected frequencies at latitudes in the subtropical/warm temperate zone between 26 and 35 degrees. Most species occurred at expected frequencies in the lower latitudes, and below expected frequencies in latitudes greater than 35 degrees.

Soil is presumed to be a major source of inoculum for Aspergillus flavus which contaminates cottonseed and produces the potent carcinogen, aflatoxin. Little is known about the mycoflora of the low desert soils of cotton fields where aflatoxin is a chronic problem. In this study, soils from cotton fields in southwestern Arizona and south-eastern California were assayed for filamentous fungi. Forty-two taxa, predominantly in the genera Aspergillus, Penicillium and Fusarium, were isolated. To determine whether or not compounds produced by these fungi could be potential inhibitors of A. flavus, extracts of strains of each taxon were tested for their ability to inhibit growth of A. flavus. Twelve taxa produced compounds inhibitory to A. flavus, including several strains of Fusarium solani, Penicillium vinaceum and Aspergillus auricomus.

Racemic gossypol and its related derivatives gossypolone and apogossypolone demonstrated significant growth inhibition against a diverse collection of filamentous fungi that included Aspergillus flavus, Aspergillus parasiticus, Aspergillus alliaceus, Aspergillus fumigatus, Fusarium graminearum, Fusarium moniliforme, Penicillium chrysogenum, Penicillium corylophilum, and Stachybotrys atra. The compounds were tested in a Czapek agar medium at a concentration of 100 μg/mL. Racemic gossypol and apogossypolone inhibited growth by up to 95%, whereas gossypolone effected 100% growth inhibition in all fungal isolates tested except A. flavus. Growth inhibition was variable during the observed time period for all tested fungi capable of growth in these treatment conditions. Gossypolone demonstrated significant aflatoxin biosynthesis inhibition in A. flavus AF13 (B(1), 76% inhibition). Apogossypolone was the most potent aflatoxin inhibitor, showing greater than 90% inhibition against A. flavus and greater than 65% inhibition against A. parasiticus (B(1), 67%; G(1), 68%). Gossypol was an ineffectual inhibitor of aflatoxin biosynthesis in both A. flavus and A. parasiticus. Both gossypol and apogossypolone demonstrated significant inhibition of ochratoxin A production (47%; 91%, respectively) in cultures of A. alliaceus.

Iturin A, a peptidolipid produced byBacillus subtilis, inhibits growth of a large number of fungi. In this study, the effects of iturin A were evaluated on nine isolates ofA. flavus and seven isolates ofA. parasiticus in liquid shake culture. The mycelial dry weight of theA. flavus isolates was not significantly influenced by iturin A, however, there was a significant reduction in mycelial dry weight for two of theA. parasiticus isolates. Aflatoxin production was significantly reduced in five of theA. flavus isolates and three of the six aflatoxigenicA. parasiticus isolates. For the other seven isolates, aflatoxin levels were either unchanged or significantly increased in the presence of iturin A. These results indicate that iturin A does not consistently reduce growth or aflatoxin production of these fungi in pure culture.

Twenty-four species placed in Aspergillus section Versicolores were examined for macromorphological and micromorphological characteristics. Based on these characters, an identification key to the species was created. A cluster analysis of the data indicated that five of the species, A. silvaticus, A. speluneus, A. floriformis, A. pseudodeflectus, and A. pulvinus were only weakly aligned with other members of the section.

Based on counts of Aspergillus species reported in over 250 studies of microfungi from soils and litter, chi-square analyses were conducted on species occurrence in five biomes and five latitude ranges to determine variations from expected distributions. There was no overall trend in distribution of the members of the entire genus by biome, however, individual sections of the genus appeared to have distinct distribution patterns. Most members of sections Aspergillus, Nidulantes, Flavipedes and Circumdati occurred at greater than expected frequencies in desert soils. There was no distinct pattern of species occurrence for forest, wetland, or cultivated soils, although members of section Nidulantes were quite rare in cultivated soils. Most species occurred at or below expected frequencies in grassland soils. Members of the genus tended to occur at greater than expected frequencies at latitudes in the subtropical/warm temperate zone between 26 and 35 degrees. Most species occurred at expected frequencies in the lower latitudes, and below expected frequencies in latitudes greater than 35 degrees.

Aspergillus ochraceoroseus produces the yellow-gold conidia and other characteristics of Aspergillus subgenus Circumdati section Circumdati. However, this species produces aflatoxin, a secondary metabolite characteristic of some members of subgenus Circumdati section Flavi and sterigmatocystin, a related secondary metabolite usually associated with subgenus Nidulantes sections Nidulantes and Versicolores, as well as members of several other genera. Our morphological data support the placement of A. ochraceoroseus in subgenus Circumdati. Sequence data from A. ochraceoroseus aflatoxin and sterigmatocystin genes aflR and nor-1/stcE, as well as 5.8S ITS and beta tubulin genes, were compared to those of aspergilli in sections Circumdati, Flavi, Nidulantes and Versicolores. In the sequence comparisons, A. ochraceoroseus was related more closely to the species in subgenus Nidulantes than to species from subgenus Circumdati.

SUMMARY Aspergillus flavus is an opportunistic pathogen of crops. It is important because it produces aflatoxin as a secondary metabolite in the seeds of a number of crops both before and after harvest. Aflatoxin is a potent carcinogen that is highly regulated in most countries. In the field, aflatoxin is associated with drought-stressed oilseed crops including maize, peanut, cottonseed and tree nuts. Under the right conditions, the fungus will grow and produce aflatoxin in almost any stored crop seed. In storage, aflatoxin can be controlled by maintaining available moisture at levels below that which will support growth of A. flavus. A number of field control measures are being utilized or explored, including: modification of cultural practices; development of resistant crops through molecular and proteomic techniques; competitive exclusion using strains that do not produce aflatoxin; and development of field treatments that would block aflatoxin production. Taxonomy: Aspergillus flavus Link (teleomorph unknown) kingdom Fungi, phyllum Ascomycota, order Eurotiales, class Eurotiomycetes, family Trichocomaceae, genus Aspergillus, species flavus. Host range: Aspergillus flavus has a broad host range as an opportunistic pathogen/saprobe. It is an extremely common soil fungus. The major concern with this fungus in agriculture is that it produces highly carcinogenic toxins called aflatoxins which are a health hazard to animals. In the field, A. flavus is predominantly a problem in the oilseed crops maize, peanuts, cottonseed and tree nuts. Under improper storage conditions, A. flavus is capable of growing and forming aflatoxin in almost any crop seed. It also is a pathogen of animals and insects. In humans it is predominantly an opportunistic pathogen of immunosuppressed patients. Useful websites: http://www.aspergillusflavus.org, http://www.aflatoxin.info/health.asp, plantpathology.tamu.edu/aflatoxin, http://www.aspergillus.org.uk

5 pages; International audience; This article documents the addition of 411 microsatellite marker loci and 15 pairs of Single Nucleotide Polymorphism (SNP) sequencing primers to the Molecular Ecology Resources Database. Loci were developed for the following species: Acanthopagrus schlegeli, Anopheles lesteri, Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus oryzae, Aspergillus terreus, Branchiostoma japonicum, Branchiostoma belcheri, Colias behrii, Coryphopterus personatus, Cynogolssus semilaevis, Cynoglossus semilaevis, Dendrobium officinale, Dendrobium officinale, Dysoxylum malabaricum, Metrioptera roeselii, Myrmeciza exsul, Ochotona thibetana, Neosartorya fischeri, Nothofagus pumilio, Onychodactylus fischeri, Phoenicopterus roseus, Salvia officinalis L., Scylla paramamosain, Silene latifo, Sula sula, and Vulpes vulpes. These loci were cross-tested on the following species: Aspergillus giganteus, Colias pelidne, Colias interior, Colias meadii, Colias eurytheme, Coryphopterus lipernes, Coryphopterus glaucofrenum, Coryphopterus eidolon, Gnatholepis thompsoni, Elacatinus evelynae, Dendrobium loddigesii Dendrobium devonianum, Dysoxylum binectariferum, Nothofagus antarctica, Nothofagus dombeyii, Nothofagus nervosa, Nothofagus obliqua, Sula nebouxii, and Sula variegata. This article also documents the addition of 39 sequencing primer pairs and 15 allele specific primers or probes for Paralithodes camtschaticus.

This review summarizes the health aspects of the medically important fungal genus Aspergillus. The morphology and systematics of the genus are explained as well as its biogeography. Major mycotoxins, the aspergilli that produce them, affected crops, and symptoms of the toxicoses are summarized, as are the major mycoses caused by aspergilli. The current status of the relationship between Aspergillus in the indoor environment and health issues are discussed.

Summary This first study by the Subcommission on Penicillium and Aspergillus Systematics is concerned with the status of the closely related species Penicillium glabrum, P. spinulosum, P. purpurescens, and P. montanense. Thirty five isolates tentatively identified as these species were studied as unknowns by the eight members of the Subcommission. Characters studied included traditional morphology and gross physiology plus secondary metabolite profiles, isoenzyme patterns and DNA restriction enzyme fragment length polymorphism. The various approaches provided consistent identifications for most isolates, indicating that the four species could be separated satisfactorily and should be maintained. Statistical models based on morphological and gross physiological data indicated that five characters were of value in distinguishing the species. These were conidial wall texture, colony diameters on Czapek yeast extract agar and 25% glycerol nitrate agar, phialide width and vesicle diameter. Practical aspects of differentiating these species are also discussed.

Cottonseeds having fluorescent fibers were harvested from fields in Arizona and examined utilizing light microscopy and transmission electron microscopy. The occurrence of fluorescent fibers indicated that seeds had been infected by Aspergillus flavus during development. Presence of A. flavus was verified by plating portions of seeds with fluorescent fibers. Hyphae, conidial heads, and conidia were identified readily in differentially-stained cotyledon tissue processed for light microscopy. Utilization of transmission electron microscopy permitted observations on lignified seed coats and cotyledons of mature cottonseeds. Hyphae were located throughout the cotyledon and in the nonlignified layers of the seed coat. The identification of hyphae in cross sections of vessel elements within the seed coat provided ultrastructural evidence supporting the hypothesis that A. flavus may enter seeds via the vascular tissue. Controls for the microscopy studies included observations on cottonseeds with no visual signs of infection and on laboratory-grown cultures of A. flavus. These observations demonstrated that the hyphae localized within fluorescent seeds had features characteristic of A. flavus and that fungal-like structures do not occur within uninfected seeds.

There is a large and rapidly growing market for fresh-cut fruit. Microbial volatile organic compounds indicate the presence of fungal or bacterial contamination in fruit. In order to determine whether microbial volatile organic compounds can be used to detect contamination before fruit becomes unmarketable, pieces of cantaloupe, apple, pineapple, and orange were inoculated with a variety of fungal species, incubated at 25 degrees C, then sealed in glass vials. The volatiles were extracted by headspace solid-phase microextraction and analyzed by gas chromatography-mass spectrometry. Forty-five compounds were identified that might serve as unique identifiers of fungal contamination. Fungal contamination can be detected as early as 24 h after inoculation.

Recent concerns over the possible use of airborne biological particles as weapons of mass destruction have significantly increased the attention that researchers are giving to this threat. The size of these particles, ranging from a fraction of a micrometer to several tens of micrometers, allows them to travel over long distances before settling out of the airstreams carrying these particles. Furthermore, the odd shapes of many of these particles along with uncertainties about their light scattering characteristics make detection and tracking quite a challenge. In the present paper, results are reported on the visualization of airborne biological particles using two-dimensional particle image velocimetry (PIV). These initial results show the utility of PIV in illuminating and tracking airborne biological particles. A compressed air nebulizer is used to aerosolize the biological particles inside a Plexiglas test section. The biological particles prepared for the nebulizer are first inoculated and cultured onto agar media, gypsum board, and acoustic ceiling tile to achieve an abundant growth of spores. A colloidal suspension of biological particles is then made using sterilized, de-ionized water and a mild surfactant to de-agglomerate the biological particles in the suspension. The concentration of biological particles in the colloidal suspension is determined using a hemacytometer. In the visualization experiments, images are captured for polystyrene latex (PSL) test particles, liquid water droplets, and spores of the fungal species Aspergillus versicolor. During the PIV system operation, two successive images are captured with a time delay of 50 μm to develop flow field velocities of the PSL test particles, liquid water droplets, and the A. versicolor spores.

The removal of many synthetic fungicides from the market has created a demand for new, environmentally safe fungicides. Iturin A, a cyclic lipopeptide produced by Bacillus subtilis, has strong antifungal properties and low mammalian toxicity. To determine the efficacy of this compound as a potential fungicide on stored feed grains, lots of corn, peanuts and cottonseed were treated with varying concentrations of iturin A. The mycoflora of treated seed was assayed along with that of untreated seed and seed treated with fungicides used commercially for planting seed. Fungal species varied considerably in their sensitivity to iturin A. Significant reductions in total mycoflora occurred in most seed lots tested at iturin A concentrations of 50 to 100 ppm.

Bacillus subtilis produces peptidolipid compounds of the iturin group that have been shown to have antifungal properties, but not all fungal species are sensitive to these compounds. In this study, the activity of iturin A, produced by B. subtilis strain B-3, was tested. Paper disks impregnated with various concentrations of iturin A were placed on agar plates seeded with conidia of toxigenic species of Fusarium, Gerlacia, Penicillium or Aspergillus. Most isolates were inhibited at iturin A concentrations as low as 4 micrograms/disk. Penicillium italicum, P. viridicatum, A. ochraceus and A. versicolor were most strongly inhibited by the iturin whereas P. citrinum and A. parasiticus were least sensitive to iturin A.

In greenhouse and field studies, cotton ( Gossypium hirsutum L.) flowers were inoculated with Aspergillus flavus at the involucral nectaries. Bolls developing from early-season flowers had significantly higher percentages of A. flavus -infected seed than did bolls from flowers formed later in the season. Seeds from bolls inoculated 2 weeks after anthesis had the same infection levels as those from flowers inoculated at anthesis. These results indicate that early-season flowers are predisposed to A. flavus infection and that the degree of susceptibility at anthesis is retained through early boll development.

Aspergillus flavus and Aspergillus oryzae are difficult to differentiate using standard morphologically based characteristics. This survey, using several restriction enzymes, showed that Sma I digestions of total DNA could be used to differentiate these two closely related species of Aspergillus . A different electrophoretic pattern was associated with each of the two species, while within a species the pattern remained constant. CsCl banding of DNA from one isolate of each species indicated the DNA that produced the species-specific pattern was associated with the nuclear DNA fraction.

Twenty-seven mature cotton bolls with Aspergillus flavus Link colonies naturally occurring on the surface of the boll or lint were collected in the field in Arizona along with their subtending stems and peduncles. Bolls inoculated through the carpel wall 30 days after anthesis were allowed to mature in the field and were collected in the same manner. The seed and stem and peduncle sections of each boll were surface-sterilized, plated on agar media and observed for A. flavus. Seventy-eight percent of the naturally contaminated bolls with A. flavus in the seed also had the fungus in the stem and peduncle, whereas only 31% of the naturally contaminated bolls with no A. flavus in the seed had the fungus in the stem or peduncle. This difference was significant (P = 0.0125), indicating a positive relationship between seed infection and stem and peduncle infection. All of the bolls inoculated through the carpel wall had A. flavus in the seed, but only 11% of the stem and peduncle sections were infected, indicating that the fungus does not readily grow downward from the boll into the supporting stem or peduncle. This unidirectional pattern of movement (upward) was further substantiated in greenhouse experiments where cotton seedlings were inoculated at the cotyledonary leaf scar with A. flavus and plants were sequentially harvested, surface sterilized and plated. Aspergillus flavus was isolated from the cotyledonary leaf scar, flower buds, developing bolls, and stem sections in the upper portion of the plant. It was never isolated from roots or stem sections below the cotyledonary node, again indicating that the fungus does not readily move downward through the plant.

Cotton dust associated with high pulmonary function decrements contains relatively high levels of mannitol. In this study, cotton leaf and bract tissue and dust isolated from cotton leaf tissue were examined by optical microscopy, scanning electron microscopy, and capillary gas chromatography. Alternaria alternata, Cladosporium herbarum, Epicoccum purpurascens , and Fusarium pallidoroseum were isolated from cotton leaf dust. The fungal samples, cotton dust, and cotton leaf contained mannitol. This study demonstrates that fungi from a late-fall harvest of cotton leaf material produce mannitol and are a probable source of the mannitol found in cotton dust.

More than 150 isolates of Aspergillus flavus, A. oryzae, A. parasiticus, A. sojae and A. tamarii from collections around the world were examined independently in two laboratories as unknowns, and data assessed to evaluate a wide range of morphological characters for suitability as taxonomic criteria. Mycotoxin production was also assessed. It was concluded that conidial wall texture was the most effective criterion for distinguishing A. flavus and A. parasiticus. A. flavus has a broad interface with VA. oryzae; therefore, a combination of characters was necessary to separate these two species. Mycotoxin production correlated well with morphological speciation. Only A. flavus and A. parasiticus isolates produced B aflatoxins. Very few A. flavus isolates produced G aflatoxins whereas almost all A. parasiticus isolates did. The A. parasiticus isolates never produced cyclopiazonic acid, but some isolates of A. flavus, A. tamarii and A. oryzae did.

Delinted, surface-sterilized cotton seeds from Regional Cotton Variety Test locations in the southern USA were assessed for presence of filamentous fungi. There were no consistent differences in fungal flora among the cultivars examined. Of the thirty-seven taxa isolated, only Alternaria spp., Colletotrichum gossypii, Fusarium equiseti, and F. pallidoroseum (F. semitectum) were present in more than 10% of the seed at any sampling location in a given year. Infection levels varied greatly between locations and in different years within a location. Changes in infection levels for some of the species were attributable to precipitation and temperature variations during the growing season.

Fusarium semitectum Berkeley and Ravenel and other nonpathogenic fungi appear to contaminate immature cottonseed but cannot express themselves until immediately after seed maturation. It is hypothesized that catechin prevents fungal outgrowth and that inhibition is released when catechin is polymerized during seed maturation.

Aspergillus flavus was introduced into several natural openings in cotton plants at or before anthesis and the subsequent infection rate in the mature seed determined. Cotyledonary leaf scars of seedlings, unopened flower buds, and involucral nectaries or stigmata of newly opened flowers were inoculated with dry fungal spores. Seeds from plants inoculated at the cotyledonary node, involucral nectaries, and flower buds had significantly higher A. flavus infection rates than uninoculated control plants.

Cotton flowers in replicate plots in two fields near Phoenix, Ariz., were tagged in June at the beginning of the flowering period. Flowers or developing bolls from these tagged flowers were inoculated on the involucral (bracteal) nectaries with dry spores of Aspergillus flavus. The bolls were harvested as they matured in August, and the seeds were assessed for the presence of the fungus. The number of infected seed from flowers and bolls inoculated up to 25 days after anthesis was significantly higher than that in uninoculated controls. Seeds from bolls inoculated after 25 days postanthesis did not differ significantly from controls in degree of infection. We postulate that the sharp decline in the ability of the fungus to infect the plant and seed is a result of physical or biochemical changes in the boll as it reaches physiological maturity or biochemical changes in the entire plant as it develops.

Sixty-two Aspergillus flavus group isolates from the SRRC and FRR culture collections were studied and identified independently by the authors. Strains were grown under standard conditions and observations recorded for a variety of morphological features at various stages of colony development. Aflatoxin production and response on a special A. flavus and A. parasiticus medium were also recorded. A discriminant analysis was performed to further study species relatedness. It was concluded that A. flavus, A. parasiticus, A. sojae, A. oryzae, and A. tamarii could be reliably separated from one another by morphological and microscopic criteria. The most salient distinguishing features were: 1) colony colour; 2) conidial size and roughening; 3) vesicle diameter; 4) conidiophore length; and 5) presence of metulae.

(1989). Use of a Bleomycin-Containing Medium to Distinguish Aspergillus Parasiticus from A. Sojae. Mycologia: Vol. 81, No. 1, pp. 159-160.

Thousands of fungal species are classified as hyphomycetes. Many of these species have a significant impact on moder society as pathogens, both plant and animal, as producers ofmycotoxins, as food spoilage and fermentation organisms, and as potential candidates for manipulation in the biotechnological industries. However, with few exceptions, i.e., Aspergillus nidulans and the anamorph of Neurospora crassa, little genetic information is available on the majority of filamentous fungi. Several developmentally regulated genes have been cloned from A. nidulans including SpoCI (Timberlake and Barnard, 1981; Gwynne et al., 1984) and tubC (May et al., 1985). SpoCI is a gene cluster that codes for several poly(A) + RNAs that are preferentially expressed during conidiophore development. Beta3-tubulin (tubC) functions in the development of microtubules during conidial mitosis and is either absent or found in only trace levels in the vegetative state. The non-developmentally regulated (i.e., is present in vegetative cultures) benA gene codes for two other beta-tubulin polypeptides, betal and beta2, in A. nidulans. Except for a survey of representatives of the genus Aspergillus (Mullaney and Klich, 1987), no information exists regarding the analogous genes in other filamentous fungi. The widespread distribution of similar genes outside the aspergilli would enhance the use of A. nidulans as a model to study other fungal species. Such a model would facilitate developmental regulation studies in many economically important hyphomycetes. As an initial step in determining if other filamentous fungi have similar developmental genes to those of A. nidulans, several hyphomycetes