Production of a Fungistat and the Role of Fungi during Germination of Digitalis purpurea L.cv. Gloxiniaflora Seeds

The seed coats of Digitalis purpurea L. cv. Gloxiniaflora werecultured for indigenous fungi. Alternaria alternata (Fries)was identified as the sole fungus on seeds freshly harvestedfrom unopened capsules, whereas A. alternata, Rhizopus arrhizus(Fischer), Penicillium sp. and other fungi appeared on storedseeds. The appearance of fungi in seed cultures was seasonal,being more frequent in winter and early spring than in summerand autumn. Alternaria grew well on autoclaved seeds, on dehiscentseed coats, or on seed coats separated from the embryos of ungerminatedseeds. Rhizopus did not grow on these but grew weakly on theculture medium from viable seeds. A. alternata appears to functionas a degradation agent for the seed coat subsequent to germination.Neither fungus was found to be essential to germination of Digitalisseeds. Bioassay of the culture medium from germinating seedsshowed that a fungistat effective against both Alternaria andRhizopus is produced coincident with germination. Based on chromatographicanalysis, the fungistat appears to be a cardenolide. Alternaria alternata (Fries), cardenolides, Digitalis purpurea L. cv, Gloxiniaflora, fungistats, germination, Rhizopus arrhizus (Fischer)     

Preharvest aflatoxin contamination of maize inoculated withAspergillus flavus andFusarium moniliforme

A two-year factorial experiment was utilized to test plants field-inoculated singly and in combination withAspergillus flavus andFusarium moniliforme. Pinbar inoculations were made through the husks with conidial suspensions, and 10-ear maize samples were harvested at 60 days post-silking for aflatoxin determinations. When ears were inoculated with both fungi simultaneously,F. moniliforme reduced aflatoxin formation byA. flavus isolate NRRL 3357 by approximately two-thirds.F. moniliforme had no significant effect on naturally occurring aflatoxin contamination byA. flavus. This may be due to the timing of infection by both fungi in the field. In nature,A. flavus andF. moniliforme respond differently to the environment, offering one explanation of whyF. moniliforme did not measurably affect the other fungus.     

Seed-borne fungi in domestic bird feed in Saudi Arabia

Seed-borne fungi from seeds of barley, canary, maize, oats, rice, rapeseed, red millet, sunflower, white millet and yellow millet were investigated. These seeds are available in the market as bird feed, supplied by companies in various countries as healthy and nutritious food for domestic birds. A total of 81 fungal species were isolated. The genusAspergillus was predominant with 23 species, followed byPenicillium with 18,Ulocladium with 7,Alternaria with 6 andChaetomium, Curvularia andDrechslera with 5 species each. Other genera isolated wereAureobasidium, Cladosporium, Fusarium, Mucor, Rhizopus andSyncephalastrum.     

Studies on seed-borne fungi of wheat in seed health testing programme

Seed mycoflora associated with wheat was studied on different media with a particular reference to Blotter and potato dextrose agar (PDA) procedures of ISTA. Seed-borne fungi, viz. Fusarium moniliforme, Rhizopus spp., Mucor spp., Alternaria alternata, Aspergillus niger, Aspergillus flavus, Curvularia lunata, Drechslera spp, Alternaria spp. and Penicillium spp., were isolated from the variety HD264. Blotter method was found to be the best media for the isolation of mycoflora whether borne externally or internally. Total number and frequency of occurrence of fungi were recorded. The effect of seed treatment with different chemicals and eco-friendly botanicals was analysed on germination, and growth, better percentage of seed germination and reduction in fungal pathogen were due to biochemical seed treatment.     

The antibiotic effect of culture filtrates of some soil fungi on rhizobial growth in cultures

Summary Four strains ofRhizobium sp. from peanut (Arachis hypogaea L.) root nodules were tested for their sensitivity to metabolites (culture filtrates) of more than ten common soil inhabiting fungi, in yeast extract mannitol agar and broth cultures. Among the rhizobial strains tested strain CB-530, BU-1 and BU-2 were not sensitive to metabolites ofMyrothecium roridum andFusarium moniliforme whereas CB-1024 was sensitive. Culture filtrates ofGliocladium roseum, Thielavia basicola andDrechslera pedicellata inhibited the strains CB-530 and BU-2 but not CB-1024. Most of the soil inhabiting fungi tested were inhibitory to rhizobial growthin vitro and very few were stimulatory, their effect in either case being strain specific.     

Fungi,Aflatoxins, Fumonisin B<Subscript>l</Subscript> and Zearalenone Contaminating Sorghum-based Traditional Malt,Wort and Beer in Botswana

Brewing and consumption of traditional beer have social–economic significance in most African countries including Botswana. Traditional sorghum malt, wort, and beer samples were collected from three villages around Gaborone, Botswana. Forty-six malt samples were analyzed for fungi on three different media and developing colonies were subcultured for identification. Rhizopus, Fusarium, Mucor, and Aspergillus were the most common genera isolated. Out of the 46 malt samples, 72% contained Rhizopus stolonifer, 63% Fusarium verticillioides (syn. Fusarium moniliforme), and 37% Aspergillus flavus. Although Aspergillus flavus was isolated from malt samples, aflatoxins (B₁, B₂, G₁, and G₂) were not detected in any of the samples analyzed. When the malt, wort, and beer samples were analyzed for fumonisin B_l and zearalenone, fumonisin B₁ was detected in 3 malt samples, with concentrations ranging from 47 to 1316 μg/kg, while zearalenone was detected in 56%, 48% and 48% of the malt, wort and beer samples, respectively. Zearalenone concentration in samples ranged from 102 to 2213 μg/kg in malt, 26 to 285 μg/l in wort and 20 to 201 μg/l, in beer. Zearalenone carry-over from wort to beer ranged from 23 to 403%. Therefore, although aflatoxins and fumonisin B₁ do not appear to be major contaminants, zearalenone is common and could pose a potential problem in traditional beer in Botswana.     

Prevalence and control of wheat (Triticum aestivum L.) seed borne fungi in farmer-saved seeds

High incidence of diseases associated with the use of seeds saved from previous harvests as desire of maintaining local varieties with special attributes is of increased concern in wheat industry worldwide. Prevalent of seed-borne fungi in farmer-saved seeds and seed dressing fungicides to prevent infection from seeds to seedlings was studied in Northern Tanzania. One hundred and thirty five untreated farmer-saved seed lots were collected randomly from farmers. Alternaria alternata, Bipolaris sorokiniana, Drechslera tritici, Fusarium graminearum, Fusarium moniliforme, Aspergillus flavus, Cladosporium sphaerospermum, Epicoccum purpurascens, Pyricularia oryzae and Penicillium corylophilum were fungi isolated in farmer-saved seeds. Mean seed infection was 29% causing average grain yield loss of 1.2 mt/ha^?1. Seed dressing with Metalaxy plus (Methyl carboxaitide), Mancozeb (Manganase-zinc salt) and Baytan (Chlorophenoxy ethanol) increased seed germination by 14, 13 and 17%, respectively, and grain yield by 28, 20 and 18%, respectively. Farmer-saved seeds were heavily infected by fungi with low grain yield performance.     

The mycoflora and incidence of aflatoxin,zearalenone and sterigmatocystin in dairy feed and forage samples from Eastern India and Bangladesh

The mycoflora of a range of mixed ruminant feed and forage samples collected in Eastern India and Bangladesh were determined. Most fungi isolated from the samples were those associated with low moisture contents. Several potentially mycotoxigenic fungi and thermotolerant species were present particularly in the rice straw. The most frequently isolated fungi were: Aspergillus versicolor (Vuill.) Tiraboschi, Penicillium citrinum Thorn, Eurotium species, Wallemia sebi (Fr.) v. Arx, Aspergillus penicilloides Speg. and yeasts. Fusarium moniliforme Sheldon and Cladosporium species were prevalent in straw but not other feeds. Levels of certain individual fungi, particularly thermotolerant types, were higher in Bangladesh rice straw although overall counts were similar to those of the Indian samples. Mycotoxin analyses for aflatoxins, zearalenone and sterigmatocystin were carried out. Aflatoxins were detected in significant amounts in some feed samples from Eastern India but not in rice straw from India and Bangladesh. Zearalenone occurred in a Paspalum straw sample and three other feed samples from India and sterigmatocystin was detected in three rice straw samples from Bangladesh.     

Mycoflora and aflatoxin production in pigeon pea stored in jute sacks and iron bins

The mycoflora, moisture content and aflatoxin contamination of pigeon pea (Cajanus cajan (L.) Millisp) stored in jute sacks and iron bins were determined at monthly intervals for a year. The predominant fungi on freshly harvested seeds wereAlternaria spp.,Botryodiplodia theobromae, Fusarium spp. andPhoma spp. These fungi gradually disappeared from stored seeds with time and by 5–6 months, most were not isolated. The fungi that succeeded the initially dominant ones were mainly members of the generaAspergillus, Penicillium andRhizopus. Population of these fungi increased up to the end of one year storage. Higher incidence of mycoflora andAspergillus flavus were recorded in jute-sack samples throughout the storage period. The moisture content of stored seeds was found to fluctuate with the prevailing weather condition, being low during the dry season and slightly high during the wet season. The stored seeds were free of aflatoxins for 3 and 5 months in jute sacks and iron bins respectively. The level of aflatoxins detected in jute-sack storage system was considerably higher than that occurring in the iron bin system. Of 196 isolates ofA. flavus screened, 48% were toxigenic in liquid culture (54% from jute sacks and 41% from iron bins).     

Fungi and mycotoxins in south african maize of the 1993 crop

The most dominant fungi in South African maize for the 1993 crop wereFusarium moniliforme andFusarium subglutinans. The mycotoxins occurring with the most frequency were the fumonisins, deoxynivalenol and nivalenol. Levels of fungal infection and mycotoxins varied between white maize and yellow maize, and between the respective grades.     

Colonization ofRetama raetam seeds by fungi and their significance in seed germination

Examination by scanning electron microscopy and incubation on potato-dextrose agar medium showed that dry seeds ofRetama raetam were externally free of fungi. When planted in sandy loam soil, the seeds become colonized with eleven soil-borne fungal species. The fungi were isolated on cellulose agar, pectin agar and lignin agar media.Aspergillus flavus, A. niger, A. fumigatus, Penicillium capsolatum andFusarium oxysporum had broad occurrence and were recovered on all the three media. The production of hydrolytic enzymes by the isolated fungi depends on the substrate and species.Penicillium capsolatum, P. spinulosum andA. niger had wide enzymatic amplitude and they were able to produce cellulolytic, pectolytic and lignolytic activities on corresponding substrates as well as on seed-coat-containing media. The lignolytic activities of the isolated species exceptChaetomium bostrychodes andTrichoderma viride were enhanced by applying the seed-coat materials as C- source rather than lignin. SoakingR. raetam seeds in culture filtrates of most of the fungi grown on seed-coat-supplemented media induced a pronounced and distinct stimulating effect on seed germination. The most effective filtrates were those ofP. capsolatum, P. spinulosum andSporotrichum pulverulentum.     

Moniliformin produced by cultures of Fusarium moniliforme var. subglutinans isolated from swine feed

Feed samples from Iowa suspected of causing vomiting and enlarged vulva as well as mortalities of swine were examined for toxigenic fungi and mycotoxins Fusarium moniliforme Sheldon and F. moniliforme Sheldon var. subglutinans Wollenew. & Reink. accounted for 43% and 18.5%, respectively, of the total count of 4.75×10⁵ propagules filamentous fungi per gram of swine feed, but representatives of various Penicillium spp. and Aspergillus spp. were also found. Eight isolates of F. moniliforme var. subglutinans from the feed produced 51–540 g of moniliformin per g on cracked corn at 25°C for six weeks. Zearalenone was not detected in these corn fermentations. Eight isolates of F. moniliforme from the feed did not produce detectable amounts of either zearalenone or monoliformin on cracked corn. Moniliformin was not detected in the feed samples.     

Studies on fungi in the root region

Summary Fungal isolations were made from roots ofPhaseolus vulgaris after washing in sterile water, at monthly intervals throughout the life of the plant, and from other roots after dissection and after surface sterilization at certain plant ages only. A table is provided showing the relative importance of the most common species isolated in each of four clearly distinct microhabitats — the root surface, the cortex, the outer stele and the inner stele.Fusarium oxysporum andCylindrocarpon radicicola were the most frequently isolated fungi from the roots.Fusarium oxysporum was most abundant on young roots and seemed to be associated, particularly, with the root surface and cortical tissues.Cylindrocarpon radicicola, although common on young roots, was more abundant on older roots and was an important initial colonist of the stelar tissues. Sterile mycelia were isolated mainly from older roots and seemed to be responsible, withC. radicicola, for the initial colonization of the stele. Microscopic examination of roots showed the cortical tissues to be increasingly penetrated by fungal hyphae with plant age but extensive fungal penetration of the endodermis and stelar tissues did not occur until the plants were at least five months old.     

Effect of delayed harvest on contamination of pearl millet grain with mycotoxin-producing fungi and mycotoxins

The response to delayed harvest of fungal and mycotoxin contamination of grain of the pearl millet hybrid HGM 100 was examined in 1992 and 1993. Samples of grain were assayed from seven plantings at locations near Tifton, Georgia, USA. Grain was harvested at 30, 40, and 50 days after anthesis and evaluated for infection byFusarium species andAspergillus flavus, and mycotoxin contamination. Mean isolation frequencies ofF. semitectum (35.6%) andF. chlamydosporum (17.2%) increased linearly with delayed harvest.Fusarium moniliforme andF. equisiti were infrequently isolated (<0.5%) and did not increase in the grain when harvest was delayed. Low mean concentrations of zearalenone (0.17 ppm), nivalenol (0.42 ppm), and deoxynivalenol (0.01 ppm) were detected but were not affected by delayed harvest. Isolation frequencies ofF. chlamydosporum andF. equiseti were correlated (P=0.07) with levels of nivalenol.Aspergillus flavus was not isolated from the grain, and aflatoxin concentrations averaged 1.9 ppb.     

Screening,statistical optimized production,and application of β‐mannanase from some newly isolated fungi

Eighty‐eight fungi isolated from soil and decaying organic matter were screened for mannanolytic activity. Twenty‐eight fungi produced extracellular mannanase on locust bean gum as evidenced by zone of hydrolysis produced on mannan agar gel. Six prominent producers, including four Fusarium species namely Fusarium fusarioides NFCCI 3282, Fusarium solani NFCCI 3283, Fusarium equiseti NFCCI 3284, Fusarium moniliforme NFCCI 3287 with Cladosporium cladosporioides NFCCI 3285 and Acrophialophora levis NFCCI 3286 produced the β‐mannanase in the range of 84–140 nkat/mL. All these grew well on particulate substrates in solid‐state fermentation (SSF), producing relatively higher titers on mannan‐rich palm kernel cake (PKC) and copra meal. Two high yielding strains, F. equiseti (1747 nkat/gds) and A. levis (897 nkat/gds) were selected for statistical optimization of mannanase on PKC. Interaction of two critical solid state fermentation parameters, pH and moisture on mannanase production by these two molds was studied by response surface method. Optimized production on PKC resulted in three‐ to fourfold enhancement in enzyme yield was observed in case of F. equiseti (5945 nkat/gds) and A. levis (4726 nkat/gds). HPLC analysis of mannan hydrolysate indicated that F. equiseti and A. levis mannanase performed efficient hydrolysis of konjac gum (up to 99%) with exclusive mannooligosaccahride (DP of 4) production. A seminative SDS‐PAGE revealed that A. levis and F. solani produced three isoforms, F. moniliforme produced two isoforms while F. fusarioides, F. equiseti, and C. cladosporioides produced a single enzyme.     

Anaerobic growth ability and alcohol fermentation activity of microscopic fungi

The method proposed in this study was used to isolate fungi grown under anaerobic conditions and to reveal distinctions in their abundance and species composition in different habitats. The ability of micromycetes of different taxa to grow under anaerobic conditions and ensure alcohol fermentation was determined for a representative sample (344 strains belonging to more than 60 species). The group of fungi growing under anaerobic conditions included species with high, moderate, and low fermentation activity. The ability for anaerobic growth and fermentation depended on the taxonomic affiliation of fungi. In some cases, the expression of these characteristics depended on the habitat from which the strain was isolated. The maximum level of ethanol accumulation in culture liquid (1.2–4.7%) was detected for Absidia spinosa, Aspergillus sp. of group flavus, Aspergillus terreus, Acremonium sp., Mucor circinelloides, Mucor sp., Fusarium oxysporum, F. solani, F. sambucinum, Rhizopus arrhizus var. arrhizus, Trichoderma atroviride, and Trichoderma sp.     

Keratinophilic fungi from soil of Brittany,France

The Present work reports the study of the distribution of keratinophilic fungi isolated from 39 samples of soil collected in Brittany, France. Keratinophilic fungi were isolated from 35 (92.3%) of the samples studied. The most frequently found species wereFusarium moniliforme, Penicillium viridicatum and an unidentified species ofAcremonium. The other fungi isolated were members of the generaChrysosporium, Gliocladium, Mucor, Trichoderma andTrichophyton.     

Resistance to Fusarium verticillioides in 20 Zambian Maize Hybrids

Visual assessment of maize ears and Fusarium spp. isolation from kernels were compared to determine resistance in 20 Zambian maize hybrids. The mean percentage Fusarium spp. isolations in non-inoculated field experiments varied between years (12–62%). Symptomless infection by Fusarium spp. had domination over symptomatic. More than 95% of the Fusarium species isolated were F. vertcillioides . A disease severity index and the percentage of visibly diseased, discoloured and damaged kernels did not differentiate hybrids with respect to Fusarium spp. ear rot under natural conditions. Artificial inoculation provided a good estimate of Fusarium spp. resistance based on visual symptoms in a year of moderate disease pressure, but not in a year of high disease pressure. The percentage Fusarium spp. isolations showed significant differences between hybrids after inoculation, and it was significantly negatively correlated with the number of days from planting to midsilk. Parental line L5522 contributed to hybrid resistance to Fusarium . The hybrids MM 701-1 and MM 752 were the most resistant among the 20 hybrids.     

A review and update of animal toxicoses associated with fumonisin-contaminated feeds and production of fumonisins by Fusarium isolates

During the 1989 corn harvest season, numerous reports of equine leukoencephalomalacia (ELEM) outbreaks and a pulmonary edema (PPE) syndrome in swine from several regions of the United States were received by the National Veterinary Services Laboratories (NVSL), Ames, Iowa. Previous and concurrent research linked Fusarium moniliforme and fumonisin-contaminated feeds to both diseases. Chemical and mycological investigations revealed fumonisin B₁ (FB₁) concentrations of 20 to 360 ppm in suspect swine feeds and 8 to 117 ppm in suspect equine feeds. Nonproblem feeds contained concentrations below 8 ppm. Fusarium moniliforme and Fusarium proliferatum were isolated from both problem and nonproblem equine and swine feeds. When cultured on autoclaved corn, the F. moniliforme and F. proliferatum isolates produced respective FB₁ and fumonisin B₂ (FB₂) that range from less than 5 to more than 2450 ppm and less than 5 to more than 1000 ppm, respectively. Isolates from both problem and nonproblem feeds produced high levels (greater than 500 ppm) in culture. Reported here is a review of chemical and mycological data resulting from the study of several cases of PPE and ELEM.     

Enterobacter cloacae is an endophytic symbiont of corn

The bacteriumEnterobacter cloacae is presently used for biocontrol of postharvest diseases of fruits and vegetables and as a preplant seed treatment for suppression of damping-off. This bacterium has apparent affinities for several grass species, but it is not considered to be an endophyte. While screening corn for fungi and bacteria with potential for biocontrol of various corn diseases, the surface-sterilized kernels of one unknown Italian corn cultivar produced fungus-free corn seedlings with roots endophytically infected byE. cloacae. This paper describes the microscopic nature ofE. cloacae RRC 101 with corn, and the in vitro control ofFusarium moniliforme and other fungi with this bacterium. Light and electron microscopy determined that this isolate ofE. cloacae was biologically associated with corn seedling roots, where it was distributed intercellularly within the cortex and stele. This is a first report of a strain of this bacterium as an endophytic symbiont of roots. Following a topical application ofE. cloacae to kernels, and upon germination this bacterium readily infected roots of two other corn cultivars. The bacterium was observed within the endosperm of germinating corn seedling, but germination was not affected. Further, the bacterium was isolated from leaves and stems of 3- to 6-week-old seedlings indicating that the above ground portions of corn were also colonized. There was no evidence of damage to cells of the root during a three to four week observation period. This bacterium was antagonistic to several isolates of the corn pathogenFusarium moniliforme, and to two other species of fungi, all of which produce mycotoxins on corn.