Intradiurnal periodicity of fungal spore concentrations (Alternaria, Botrytis, Cladosporium, Didymella, Ganoderma) in Cracow, Poland

Aerobiological monitoring enables the definition of seasonal fungal spore concentrations and also intradiurnal time when the highest concentrations of spores could cause or increase allergy symptoms. These data are useful to estimate symptoms of disease, duration of infection and how advanced the illness is in people suffering from fungal allergens. The aim of the study was to compare the concentrations of fungal spores (Alternaria, Botrytis, Cladosporium, Didymella, Ganoderma) during dry and rainy periods and to analyse their intradiurnal changes. Average daily spore concentrations in dry and rainy periods were compared, using z test, separately for each taxon, season and for a combined 3-year period. Intradiurnal periodicity of fungal spore concentrations was analysed on the basis of three complementary diagrams. These spore concentrations were presented using three curves for all, dry and rainy days in 1997–1999 (April–November). The spore percentage in particular hours was normalized in relation to the daily spore sum accepted as 100%. Two further diagrams enabled the more precise analysis of the highest concentrations in dry days. Daily Botrytis and Cladosporium spore concentrations did not show significant differences between dry and rainy periods. In the case of Didymella and Ganoderma spore concentrations, there were no significant differences between both weather types in the single years, although there was a significant difference when a 3-year period was considered. The differences between daily concentrations of Alternaria spores in dry and rainy periods occurred in 1997 and in a 3-year period. Intradiurnal periodicity of spore concentrations was different for ‘dry’ and ‘wet’ fungal spores. Dry spores are released from the spore-producing parts of the fungus under conditions of decreasing humidity and increasing airflow. Examples of dry spores are those from Alternaria, Cladosporium and Botrytis. Wet spores, such as those from many Ascomycetes (Didymella) and Basidiomycetes (Ganoderma), are released into the atmosphere by processes related to humidity conditions or rain. The highest concentrations of ‘dry’ spores were observed early in the afternoon, while highest values of ‘wet’ spore concentrations occurred in the predawn hours. Statistically non-significant differences between daily spore concentrations in dry and rainy periods of single seasons were found except for Alternaria. Statistically significant differences could occur when the studied period was longer than one season (Alternaria, Didymella, Ganoderma). The highest concentrations of Alternaria, Botrytis and Cladosporium spores were recorded at noon and early in the afternoon. Concentrations of Didymella and Ganoderma spores were highest in the predawn hours.     

Effect of temperature,relative humidity and aphid developmental stage on the efficacy of the mycoinsecticide Mycotal® against Myzus persicae

The green peach aphid, Myzus persicae, is a major pest worldwide. An examination of the impact of temperature, relative humidity (RH) and developmental stages of M. persicae on the efficacy of the whitefly mycoinsecticide Mycotal®, based on Lecanicillium muscarium and the effects of infection on aphid fecundity was evaluated under controlled conditions. Although this fungus can be grown at a broad range of temperatures (15–30°C), the optimum temperature for control of M. persicae ranged between 20 and 30°C. L. muscarium had high efficacy as a microbial control agent against M. persicae between 55% and 90% RH. Total mortality of aphids treated with different spore dosages of L. muscarium varied according to the developmental stage: adults, fourth and third instar nymphs proved more susceptible than first instar nymphs. Although the fungus did not affect the rate of nymph production, the reproductive period of aphids significantly decreased with increasing the spore dosage. Thus, total fecundity of treated aphids was 22.6 ± 1.1 and 31.6 ± 2.4 offspring per adult at the medium (644 ± viable spore/mm²) and low (330 ± 40 viable spore/mm²) dosages, compared with 45.7 ± 4.3 offspring per untreated aphid. The results suggest that L. muscarium has the potential as a biological control agent of M. persicae.     

Natural Occurrence ofChondrostereum purpureumin Relation to Its Use as a Biological Control Agent in Canadian Forests

The incidence of natural fructification ofChondrostereum purpureumwas estimated quantitatively on southern Vancouver Island during two winter seasons in randomly located 1000-m²plots and compared with potential added fructification that might occur as a result of using the fungus to control stump-sprouting of hardwood weeds in young forest stands. Fructification was surveyed in forests as well as in urban or agricultural areas by estimating the surface areas of woody substrates covered with basidiocarps. In addition to random plots, estimates were made also in locations where the fungus would be expected to occur (woodpiles, silvicultural thinnings, and killed trees). Basidiocarps were found throughout the area in various types of forest cover as well as in urban or agricultural situations. The amount of added fructification through the use of the fungus as a biological control agent was determined from inoculated plots as well as from calculated stump-surface areas developed from published stand-density data. Added fructification was multiplied by a factor representing the maximum biological control frequency in order to compare added fructification with natural fructification values. From the various calculations, it was determined that the added fructification ofC. purpureumis of the same order of magnitude as naturally occurring levels or even lower. In addition, there is a distinct geographical separation between predominantly forestry and predominantly settled areas where fruit and ornamental trees are cultivated. Accordingly, it was concluded that using the fungus as a biological control agent in forestry is not likely to pose a significant threat to fruit growing and commercial forests.     

Relationship between agitation speed and the morphological characteristics of Verticillium lecanii CS-625 during spore production

Verticillium lecanii is recognized as an entomopathogenic fungus, and has high potential in the biological control of pests. In this study, it was investigated that the relationship between agitation speed in a 2.5 L stirred tank reactor (STR) at 25°C and initial pH 5.5, and the morphological characteristics of V. lecanii CS-625, such as hyphal length/width, spore length/width, and the number of tips during spore production. The agitation speed affected the hyphae patterns and the number of tips. The number of spores rapidly increased at 48 to 60 h of cultivation, and the highest spore productivity (2.5 × 10¹⁰ spore/L·h at 60 h) occurred with an agitation speed of 350 rpm and an aeration rate of 1.0 vvm. The number of tips increased in proportion to the increase in spore production during the same culture time. The highest number of tips (4.8 × 10⁸ tipJ.mL) was obtained at 72 h of cultivation. The shortest mean spore length (2.8 μm) was obtained at 60 h of cultivation. Therefore, it was determined that the increased number of tips and decreased mean spore length were closely related to the production of V. lecanii spores.     

Biocontrol of aerial plant diseases in agriculture and horticulture: current approaches and future prospects

Until recently, the majority of research on the biological control of aerial plant diseases was focused on control of bacterial pathogens. Such research led to the commercialization of the biocontrol agent Pseudomonas fluorescens A506, as BlightBan A506™, for control of fire blight of pear. In contrast, chemical fungicides typically have provided adequate control of most foliar fungal pathogens. However, fungicide resistance problems, concerns regarding pesticide residues and revocation of registration of certain widely used fungicides have led to increased activity in the development of biocontrol agents of foliar fungal pathogens. Much of this activity has centered around the use of Trichoderma spp and Gliocladium spp to control Botrytis cinerea on grape and strawberry. The biocontrol agent Trichoderma harzianum T39 is commercially available in Israel, as Trichodex ™, for control of grey mold in grapes and may soon be registered for use in the US. Also targeted primarily against a foliar disease of grapes, in this case powdery mildew caused by Uncinula necator, is the biocontrol agent Ampelomyces quisqualis AQ10, marketed as AQ^{10  TM} biofungicide. Another promising development in the area of foliar disease control, though one which is not yet commercialized, is the use of rhizobacteria as seed treatments to induce systemic resistance in the host plant, a strategy which can protect the plant against a range of bacterial and fungal pathogens. Received 06 February 1997/ Accepted in revised form 05 June 1997     

Fate and role of peroxisomes during the life cycle of the yeast Saccharomyces cerevisiae: inheritance of peroxisomes during meiosis

 Sporulation in the yeast Saccharomyces cerevisiae is a meiotic developmental process that occurs in MAT a/MATα heterozygotes in response to nutrient deprivation. Here, the fate and role of peroxisomes during sporulation and germination has been examined by a combination of immunoelectron microscopy and the use of pex mutants defective in peroxisomal functions. Using a green fluorescent protein probe targeted to peroxisomes we show that peroxisomes are inherited through meiosis and that they do not increase in number either during sporulation or spore germination. In addition, there is no requirement for peroxisome degradation prior to spore packaging. Unlike the situation in filamentous fungi, peroxisomes do not proliferate during the yeast life cycle. Functional peroxisomes are dispensable for efficient meiotic development on acetate medium since homozygous Δpex6 diploids sporulated well and produced mature spores that were resistant to diethyl ether. Like haploids, diploid cells can proliferate their peroxisomes in response to oleate as sole carbon source in liquid medium, but under these conditions they do not sporulate. On solid oleate medium, homozygous pex5,Δpex6, and pex7 cells were unable to sporulate efficiently, whereas the wild type was. The results presented here are discussed in terms of the transmission of organelles to progeny cells. Accepted: 19 December 1997     

Microbial selection strategies that enhance the likelihood of developing commercial biological control products

Research interest in utilizing microorganisms to create a microbial environment suppressive to plant pathogens has increased exponentially in recent years. Despite intense interest in developing biological control agents, relatively few antagonists have achieved ‘commercial product’ status. The fact that such a small proportion of active laboratory antagonists are developed into biological control products is partly due to several features common to microbial selection strategies that are widely utilized to obtain putative biological control agents: (a) relatively few candidate microorganisms are tested; (b) microbes are selected based on the results of an assay that does not replicate field conditions; and (c) the amenability of microbes to commercial development is excluded as a selection criterion. Selection strategies that enhance the likelihood of developing commercial biological control products are described. These include making appropriate choices regarding the pathosystem for biological control, the method of microbe isolation, and the method of isolate characterization and performance evaluation. A model system of developing a biological control product active against Gibberella pulicaris (Fries) Sacc. (anamorph: Fusarium sambucinum Fuckel), the primary causal agent of Fusarium dry rot of stored potatoes, is used to illustrate the proposed selection strategy concepts. The crucial importance and methodology is described, of selecting strains with enhanced potential for commercial development based on a strain exhibiting both favorable growth kinetics and bioefficacy when grown in commercially feasible liquid media. Received 06 February 1997/ Accepted in revised form 29 May 1997     

Aerobiology in plant protection

Summary The control of phytopathogenic fungi in agriculture requires sometimes excessive chemical treatments because, in many cases, reliable monitoring systems in order to assess the real risk for the cultivations are not available.Therefore, the likelihood to assess the presence and the amount of phytophatogenic fungus propagules in the air with a spore-trap has been investigated in order to evaluate their correlation with the climate and the disease occurrence. Four phytopathogenic fungi, namelyVenturia inequalis, Phytophthora infestans, Cercospora beticola andDiaporthe phaseolorum var.caulivora were taken into consideration.Practical use of airborne spore monitoring in a disease control strategy proved to be useful only forVenturia inequalis andPhytophthora infestans causal agent of apple scab and tomato late blight respectively. For the latter, the utilisation of an aerobiological monitoring system for the disease prediction can be assumed.     

Sedimentation velocity of myxomycete spores

Sedimentation velocities were measured for seven myxomycete species and one fungus. Values for these first measurements for Myxomycetes were fitted with the formula of Stoke’s law for the terminal velocity of small spherical bodies in air. The obtained correlation coefficient of R=0.939 indicates that sedimentation velocities of myxomycete spores follow Stoke’s law well. With spore density as a parameter, the fit estimated a mean density of 0.74 g/cm³ for air-dried spores. The importance of the stalked spore case as well as the spore diameter for dispersal abilities of Myxomycetes is discussed.     

Einsatz von V. lecanii als biologisches Schädlingsbekämpfungsmittel gegen den Bohnenrostpilz U. appendiculatus var. appendiculatus im Feld und im Gewächshaus

The use of Verticillium lecanii as a biological control agent against the bean rust fungus Uromyces appendiculatus var. appendiculatus in the field and in the glasshouse The deuteromycete V. lecanii parasites uredo- and teliospores of the bean-rust-fungus U. appendiculatus var. appendiculatus. We investigated the conditions for the use of the hyperparasite as biological control agent in the field and in glasshouses. The growth rate of the hyperparasite was 0,3 cm per day at 25 °C. Under suitable conditions in the lab (25 °C, 100 % r. h.) it took about 20 days to invade 100 % of uredospores and 65 % of teliospores. We failed to prevent the spread of bean-rust-fungus spores in the field, but we succeeded in the glasshouse by 68 %, compared to the untreated controls, using the hyperparasite V. lecanii as biological control agent.     

Control of the bean rust fungus <Emphasis Type="Italic">Uromyces appendiculatus</Emphasis> by means of <Emphasis Type="Italic">Trichoderma harzianum</Emphasis>: leaf disc assays on the antibiotic effect of spore suspensions and culture filtrates

Several species of the fungal genus Trichoderma act as antagonists of other fungi. A number of strains from the Trichoderma species T. harzianum Rifai are used as biological control agents for the control of soilborne as well as foliar plant pathogens. Six T. harzianum strains, five of them isolated from commercial preparations, were evaluated for their capability to control the bean rust fungus Uromyces appendiculatus (Pers. ex Pers.) Unger. Different kinds of leaf disc assays were performed with conidial spore suspensions and sterile culture filtrates of the T. harzianum strains. Great differences were observed concerning the efficacy of the Trichoderma strains to reduce the number of the uredial pustules developing after rust inoculation which followed the application of the particular Trichoderma strains. Efficacy values ranged from 1 to over 50%. Increasing spore or culture filtrate concentrations of the two most effective isolates T12 and T_U led to decreases in the number of developing uredial pustules. Culture filtrate applications had a protective but no curative effect. T12 spore suspensions maintained their disease reducing activity even when autoclaved. This and some other evidence for an antibiotic interaction between T. harzianum and U. appendiculatus are discussed. Handling Editor: Reijo Karjalainen.     

Statistical model based optimization of spore production by solid-state culture of Beauveria bassiana

A significant enhancement of spore production by solid-state culture of Beauveria bassiana was achieved via a statistical modeling and optimization approach. A two-level Plackett–Burman design was implemented to screen the significant medium components for spore production. Among the six variables tested, sucrose, NaNO₃, and H₃BO₃ were identified as the most significant factors (confidence levels above 95%) for spore production. The steepest ascent experiments were adopted to determine the optimal region of the medium composition. The developed second-order polynomial model predicted that the maximal spore production was 6.24×10⁹/g with the following optimal culture conditions: sucrose 34.7 g/L, NaNO₃ 52.8 g/L, and H₃BO₃ 79.6 µg/L. Finally the verification experiments were carried out and the maximal spore production reached 6.221×10⁹/g at the above optimal conditions. The spore production in the verification experiments was very close to that predicted by the model, indicating that the constructed second-order model had superior modeling and optimization ability and can be applied in the optimal spore production of Beauveria bassiana YP9 as a biological control agent.     

Muscodor albus and its biological promise

We have found a novel fungal genus that produces extremely bioactive volatile organic compounds (VOCs). This fungal isolate was initially discovered as an endophyte in Cinnamomum zeylanicum in a botanical garden in Honduras. This endophytic fungus, Muscodor albus, produces a mixture of VOCs that are lethal to a wide variety of plant and human pathogenic fungi and bacteria. It is also effective against nematodes and certain insects. The mixture of VOCs has been analyzed using GC/MS and consists primarily of various alcohols, acids, esters, ketones, and lipids. Final verification of the identity of the VOCs was carried out by using artificial mixtures of the putatively identified compounds and showing that the artificial mixture possessed the identical retention times and mass spectral qualities as those of the fungal derived substances. Artificial mixtures of the VOCs nicely mimicked the biological effects of the fungal VOCs when tested against a wide range of fungal and bacterial pathogens. Potential applications for “mycofumigation” by M. albus are currently being investigated and include uses for treating various plant parts, and human wastes. Another promising option includes its use to replace methyl bromide fumigation as a means to control soil-borne plant diseases.     

PEG-mediated plastid transformation in higher plants

Summary Plastids are surrounded by an envelope consisting of a double membrane. This barrier has to be penetrated or overcome by the DNA when transforming the plastome. Both the biolistic and polyethylene glycol-mediated transformation techniques accomplish this task, albeit by different mechanisms. We were the first laboratory to successfully use the polyethylene glycol (PEG)-method for plastid transformation, yet we use the particle gun when appropriate. In this report we compare the two methods and discuss their shortcomings and advantages. Plastid transformations with various constructs, mainly using theaadA gene as a selective marker, were performed. We point to potential problems likely to be encountered during the transformation and selection processes and offer possibilities for improvement. We give further examples of the successful application of plastome transformation and show its merits in addressing biological questions concerning the elucidation of plastid sequences of unknown function and the control of plastid gene expression. Based on a presentation in the symposium “Organelle Transformation” during the 1997 SIVB Congress held in Washington, DC June 14–18, 1997. An erratum to this article is available at .     

A new strain of the rust fungus Puccinia chondrillina for biological control of skeleton weed in Australia

A strain of the rust fungus Puccinia chondrillina introduced into Australia in 1971 for the biological control of skeleton weed, Chondrilla juncea, attacked only the narrow-leaf form of this weed. Studies were, therefore, undertaken in the Mediterranean region to discover strains of the rust which will attack the other two Australian forms of skeleton weed, intermediate- and broad-leaf. Several strains of the rust were found to be highly virulent against the intermediate form. One of these strains from Manisa, western Turkey, was both highly specific to its host and safe for use as a biological control agent. This strain has recently been introduced into Australia for the control of the intermediate form of Chondrilla.     

Antibiotics and enzymes produced by the biocontrol agent Streptomyces violaceusniger YCED-9

Streptomyces violaceusniger strain YCED-9 is an antifungal biocontrol agent antagonistic to many different classes of plant pathogenic fungi. We discovered that strain YCED-9 produces three antimicrobial compounds with antifungal activity. These compounds were purified and identified, and included: AFA (Anti-Fusarium Activity), a fungicidal complex of polyene-like compounds similar to guanidylfungin A and active against most fungi except oomycetes; nigericin, a fungistatic polyether; and geldanamycin, a benzoquinoid polyketide highly inhibitory of mycelial growth of Pythium and Phytophthora spp. Antimicrobial assays were developed to estimate the production of each antibiotic independently. Medium composition had differential effects on the production of each metabolite. The hydrolytic enzymes chitinase and β-1,3-glucanase are also produced under induction by colloidal chitin and laminarin, respectively. Fungal cell walls induced the production of both enzymes. A potential for biological control of diseases caused by P. infestans was also suggested by strain YCED-9’s strong in vitro antagonism towards pathogenic isolates of this fungus. Received 27 October 1997/ Accepted in revised form 8 June 1998     

Biological control of oliver scale,Parlatoria oleae (Colvée), in a deciduous fruit Orchard in California

Conclusions Data obtained during the years 1960–1966 show conclusively that the olive scale parasites,Aphytis maculicornis andCoccophagoides utilis, when well-established, are capable of fully effective control of their host on a commerciallty-grown deciduous fruit host.A. maculicornis alone, prior to the establishment ofC. utilis, failed to reduce or maintain control of scale densities. Subsequent to the colonization and establishement ofC. utilis in 1962 and its assumption of a contributing role as an agent of biological control of oliver scale, both species of parasites were able to reduce the host below economic levels within a year. Scale densities have decreased continually since 1963. Continuation of satisfactory biological control is indicated by the low densities of scales now maintained by its parasites. Judicious use of chemical treatments, both in selection and in timing of application, to control other insect and mit problems will ensure the continuing biological control of oliove scale in this deciduous orchard.     

Pathogenicity and Host Specificity of Mycovellosiella lantanae var. lantanae,a Potential Biocontrol Agent for Lantana camara in South Africa

Lantana camara is a poisonous, bushy shrub from South and Central America that has invaded much of the moist, warm sub-tropical areas of South Africa. In the last decade, evidence of a conspicuous and damaging mycobiota on lantana in the Neotropics has persuaded biocontrol researchers to consider fungi as potential biocontrol agents for this plant. Samples of diseased L. camara leaves were collected during field surveys in South, North and Central America from 1987 to 1997. Pathogenicity tests showed the leaf spot fungus, Mycovellosiella lantanae var. lantanae to be a promising biocontrol pathogen. Inoculations onto South African biotypes of L. camara and a number of closely related species within the family Verbenaceae were made under quarantine conditions to determine the pathogenicity and host specificity of the fungus. Inoculation of L. camara biotypes resulted in necrotic, grey lesions (20-60 per leaf), necrosis of flower buds and stalks, as well as defoliation of certain biotypes after three weeks. None of the other plant species tested showed any disease symptoms. This indicates a very restricted host range, making this pathogen a suitable candidate for application as a biological control agent. It is expected that the fungus will reduce the vigour and reproductive potential of L. camara. Due to the variation in virulence of M. lantanae var. lantanae when tested on South African lantana biotypes, release strategies may include the use of combinations of the isolates to target a wider range of lantana biotypes in the field. Permission to release M. lantanae var. lantanae in South Africa was granted in September 2001. The pathogen will be released as a classical biocontrol agent in South Africa.     

Importance of ambient saturation deficits in an epizootic of the fungus Neozygites floridana in cassava green mites (Mononychellus tanajoa)

The mite-pathogenic fungus Neozygites floridana Fisher (Entomophthorales: Neozygitaceae) is considered to have potential for the biological control of the cassava green mite, Mononychellus tanajoa (Bondar). However, its activity is sporadic and laboratory data suggest a strong dependence on night-time saturation deficits for transmission. We report on an epizootic of this fungus in a mite population in northeastern Brazil. During the epizootic, host populations appeared to be limited by a combination of the pathogen and a predatory mite Neoseiulus idaeus (Acari: Phytoseiidae). When temperatures increased, the epizootic finished and the host population began to grow. Abiotic conditions could not explain the variation in host mortality following pickup of infective propagules in this epizootic. However, night-time saturation did help to explain the variation in transmission from infective cadavers to newly killed hosts. This supports laboratory observations that horizontal transmission between hosts is determined mainly by saturation deficits, while the process of infection is little affected by abiotic conditions. A further field observation was the near-absence of resting spores in dead mites (ca. 0.1% of cadavers), suggesting that the pathogen population was unsuccessful in producing inoculum to infect future M. tanajoa populations. The implications are that this pathogen will only be effective as a biological control agent in periods of high relative humidity, and establishment in new areas may be limited by resting spore formation. This revised version was published online in July 2006 with corrections to the Cover Date.     

Investigation of factors influencing spore germination of Paenibacillus polymyxa ACCC10252 and SQR-21

Bioorganic fertilizer containing Paenibacillus polymyxa SQR-21 showed very good antagonistic activity against Fusarium oxysporum. To optimize the role of P. polymyxa SQR-21 in bioorganic fertilizer, we conducted a study of spore germination under various conditions. In this study, l-asparagine, glucose, fructose and K⁺ (AGFK), and sugars (glucose, fructose, sucrose, and lactose) plus l-alanine were evaluated to determine their ability to induce spore germination of two strains; P. polymyxa ACCC10252 and SQR-21. Spore germination was measured as a decrease in optical density at 600 nm. The effect of heat activation and germination temperature were important for germination of spores of both strains on AGFK in Tris–HCl. l-Alanine alone showed a slight increase in spore germination; however, fructose plus l-alanine significantly induced spore germination, and the maximum spore germination rate was observed with 10 mmol l⁻¹ l-alanine in the presence of 1 mmol l⁻¹ fructose in phosphate-buffered saline (PBS). In contrast, fructose plus l-alanine hardly induced spore germination in Tris–HCl; however, in addition of 10 mmol l⁻¹ NaCl into Tris–HCl, the percentages of OD₆₀₀ fall were increased by 19.6% and 24.3% for ACCC10252 and SQR-21, respectively. AGFK-induced spore germination was much more strict to germination temperature than that induced by fructose plus l-alanine. For both strains, fructose plus l-alanine-induced spore germination was not sensitive to pH. The results in this study can help to predict the effect of environmental factors and nutrients on spore germination diversity, which will be beneficial for bioorganic fertilizer storage and transportation to improve the P. polymyxa efficacy as biological control agent.