Some observations concerning microsclerotia and spore production of Mycoleptodiscus terrestris in culture

Some interesting growth characteristics of Mycoleptodiscus terrestris were discovered while evaluating the fungus as a potential bioherbicide for management of the submersed macrophyte, Hydrilla verticillata. Microsclerotia were produced readily in a liquid culture medium containing a basal salts solution supplemented with corn steep liquor powder and glucose. Dried microsclerotia were capable of germinating hyphally within 24 h and sporogenically within 72 h. Initiation of spore production was not dependent on the presence of host plant material or light.     

Effect of environmental factors on disease development caused by the fungal pathogen Plectosporium alismatis on the floating-leaf stage of starfruit (Damasonium minus), a weed of rice

Starfruit, Damasonium minus (R. Br.) Buch, is a herbicide-resistant, aquatic-weed occurring in Australian rice fields. The fungal pathogen Plectosporium alismatis (Oudem) W.M. Pitt, W. Gams & U. Braun is a potential mycoherbicide for the control of this weed. Previous work showed that the fungus can suppress the growth of starfruit juvenile plants. This paper reports the effect of environmental factors on the disease development on starfruit at the floating-leaf stage. Fungal inoculation of plants in advanced phases of floating leaf stage reduced significantly (P<0.05) the green leaf area, but only when the percentages of diseased areas were deducted. However, inoculation when the first floating leaf was emerging reduced significantly (P<0.001) both, plant dry above-ground biomass and leaf area.     

Compatible solutes of sclerotia of Mycoleptodiscus terrestris under different culture and drying conditions

Mycoleptodiscus terrestris is a plant pathogen which has been shown to be effective in controlling invasive aquatic weeds in inundative biocontrol applications. The preferred propagule for production and application is the sclerotium. In the current study, we evaluated the accumulation of carbohydrates and polyols in the sclerotia of M. terrestris under different culture and drying regimes. The carbohydrates and polyols screened for represent a class of compatible solutes found in fungi. The results show mannitol and trehalose are the major analytes identified in the sclerotia of M. terrestris. We identified higher levels of mannitol and trehalose in liquid-culture produced samples relative to solid-state produced samples. The impact of fermentation time was examined and shown to impact solute levels. In addition, the drying regime was varied to produce samples dried to different moisture contents. The experiments show greater drying led to higher mannitol levels, while trehalose levels remained constant. Rapid drying of the sclerotia in a fluidized-bed dryer also show rapid accumulation of mannitol and trehalose, which suggest the enzyme activities needed for production are readily available. The findings confirm mannitol and trehalose are important metabolites in M. terrestris and their concentrations are responsive to osmotic stress conditions.     

Development of pilot-scale fermentation and stabilisation processes for the production of microsclerotia of the entomopathogenic fungus Metarhizium brunneum strain F52

Using 100 L stirred-tank bioreactors, we evaluated the effect of fermentation parameters and drying protocols on the production and stabilisation of microsclerotia (MS) of the entomopathogenic fungus Metarhizium brunneum (formerly M. anisopliae F52). Results showed that stirred-tank bioreactors can be used to mass produce stable MS of Metarhizium and that culturing and drying protocols significantly affected MS yield and stability. Length of fermentation (4–7 days) for Metarhizium cultures had no significant impact on biomass accumulation, MS formation or the storage stability of the air-dried MS granules. Although cultures of Metarhizium grown on media with a carbon-to-nitrogen (C:N) ratio of 30:1 produced significantly more biomass when compared to cultures grown in media with a C:N ratio of 50:1, MS formation and desiccation tolerance following drying were similar. After storage for 1 year at 4°C, conidia production by air-dried MS granules from 50:1 media was significantly higher compared to MS granules from 30:1 media. The addition of diatomaceous earth (DE) to cultures of Metarhizium prior to drying at rates of 0–60 g L^?1 had no significant effect on MS desiccation tolerance but did impact conidia production. Air-dried MS granules without DE produced significantly more conidia g^?1 during the first 4 months of storage, but after 1 year, conidia production was similar regardless of DE content of the MS granule. Microsclerotial granules with higher moisture levels (2.6–5.0% w/w) produced significantly more conidia immediately after drying and MS granules with low moisture (0–2.5% w/w) produced more conidia after 12 months storage.     

Influence of photoperiod on tuber production in various races of Hydrilla (Hydrilla verticillata)

The potential for asexual reproduction throughproduction of tubers was evaluated in various races ofan International Hydrilla [Hydrilla verticillata (L.f.)Royle] germplasm collection. Interracecomparisons were made under both shortday and longdayphotoperiods in environmental growth chambers. Allthe races evaluated produced rates varying from91-7182 tubers per sq m of production under shortdayconditions. Tuber production occurred under longdayphotoperiods in several races. Under propertemperature conditions, all the monoecious racescurrently established in the U.S. appear capable oftuber production throughout the year.     

Sclerotia growth and carotenoid production of Penicillium sp PT95 during solid state fermentation of corn meal

Using corn meal as fermentation substrate, the effect of some factors, fermentation time and supplementation of saccharide and nitrogen sources as well as vegetable oil, on the sclerotia growth and carotenoid production of Penicillium sp PT95 during solid state fermentation were studied. When PT95 strain was grown on the amended medium by supplementing of 3g NaNO3, 10g maltose and 2.5g soybean oil per liter of salt solution to basal medium for 20 days, the dry sclerotia weight and carotenoid yield reached 9.70 g and 5260 g / 100 g of substrate, respectively. Without supplementation only 5.36g dry sclerotia and 2149g carotenoid / 100 g of substrate was attained. © Rapid Science Ltd. 1998     

VdMsb regulates virulence and microsclerotia production in the fungal plant pathogen Verticillium dahliae

The vascular wilt fungus Verticillium dahliae infects the roots of cotton plants and can seriously diminish the yield and quality of this and other dicotyledons. However, the key genes involved in V. dahliae infection and pathogenesis in cotton remain unclear. Msb encodes a transmembrane mucin that is highly conserved in the MAPK signal pathway. Msb has been implicated previously in pathogenicity in various aerial plant fungi. In this study, V. dahliae Msb (VdMsb) was found to be required for fungal virulence and microsclerotia production. Strains lacking VdMsb exhibited reduced conidiation and microsclerotia formation. Compared with wild-type and gene-complemented strains, the invasive growth and adhesive capacity of VdMsb deletion mutants were significantly decreased. These results suggest that VdMsb plays a role in development and virulence in V. dahliae.     

Evaluation of the bioherbicide Myrothecium verrucaria for weed control in tomato (Lycopersicon esculentum)

An isolate of the fungus Myrothecium verrucaria was evaluated for its biocontrol potential against common purslane, horse purslane, spotted spurge, and prostrate spurge, all serious weed pests in commercial tomato fields in the southeastern US. In greenhouse and field tests, M. verrucaria was highly virulent against these weeds when applied as conidial sprays formulated in 0.2% Silwet L-77 surfactant, even in the absence of dew. In field test plots naturally infested with these weeds, seedlings in the two-to-three leaf growth stage treated with M. verrucaria at 2×10⁷ conidia mL^-1 in 0.2% Silwet, exhibited leaf and stem necrosis within 24 h following inoculation, with mortality occurring within 96 h. After 7 days, M. verrucaria had killed 90-95% of both purslane species and 85-95% of both spurge species. Tomatoes that were transplanted into plots treated with M. verrucaria remained healthy and vigorous throughout the growing season. Since M. verrucaria effectively controlled several common weeds under field conditions, this fungus appears to have potential as an effective bioherbicide for pre-plant weed control in production systems with transplanted tomato.     

Leaf scarring by the weevils Neochetina eichhorniae and N. bruchi enhances infection by the fungus Cercospora piaropi on waterhyacinth, Eichhornia crassipes

Additive or synergistic effects among introduced and native insect and plant pathogen agents are necessary to achieve biological control of waterhyacinth (Eichhornia crassipes), a globally damaging aquatic weed. In field plots, plants were infested with waterhyacinth weevils (Neoechetina bruchi and N. eichhorniae) and leaves were scarred by weevil feeding. Subsequent infection by the fungal pathogen Cercospora piaropi caused necrotic lesions to form on leaves. Necrosis development was 7.5- and 10.5-fold greater in plots augmented with both weevils and C. piaropi and weevils alone, respectively, than in plots receiving only C. piaropi. Twenty-four days after weevil infestation, the percentage of laminar area covered by lesions on third-youngest and oldest live leaves was elevated 2.3–2.5-fold in plots augmented with weevils. Scar density and necrosis coverage on young leaf laminae were positively correlated, even though antipathogenic soluble peroxidases were elevated 3-fold in plots augmented with weevils alone or weevils and C. piaropi. Combined weevil and fungal augmentation decreased shoot densities and leaves per plant. In a no-choice bioassay, weevil feeding on oldest but not young leaves was reduced 44 two weeks after C. piaropi inoculation. Protein content and peroxidase activities were elevated 2–6-fold in oldest leaves three weeks after inoculation. Augmentation with both waterhyacinth weevils and C. piaropi led to the development of an additive biological control impact, mediated by one or more direct interactions between these agents, and not plant quality effects.     

Management of kudzu by the bioherbicide,Myrothecium verrucaria,herbicides and integrated control programmes

Replicated field plots were established and monitored for two years to evaluate management practices for kudzu. The bioherbicidal plant pathogen, Myrothecium verrucaria, several herbicides and a variety of integrated control programmes achieved a high level of kudzu suppression, although no system tested reliably achieved eradication in this time frame.     

The fermentation stoichiometry of Thermotoga neapolitana and influence of temperature,oxygen, and pH on hydrogen production

The hyperthermophilic bacterium, Thermotoga neapolitana, has potential for use in biological hydrogen (H₂) production. The objectives of this study were to (1) determine the fermentation stoichiometry of Thermotoga neapolitana and examine H₂ production at various growth temperatures, (2) investigate the effect of oxygen (O₂) on H₂ production, and (3) determine the cause of glucose consumption inhibition. Batch fermentation experiments were conducted at temperatures of 60, 65, 70, 77, and 85°C to determine product yield coefficients and volumetric productivity rates. Yield coefficients did not show significant changes with respect to growth temperature and the rate of H₂ production reached maximum levels in both the 77°C and 85°C experiments. The fermentation stoichiometry for T. neapolitana at 85°C was 3.8 mol H₂, 2 mol CO₂, 1.8 mol acetate, and 0.1 mol lactate produced per mol of glucose consumed. Under microaerobic conditions H₂ production did not increase when compared to anaerobic conditions, which supports other evidence in the literature that T. neapolitana does not produce H₂ through microaerobic metabolism. Glucose consumption was inhibited by a decrease in pH. When pH was adjusted with buffer addition cultures completely consumed available glucose. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

The effect of weed control practices on macroinvertebrate communities in Irish Canals

Macroinvertebrates in aquatic habitats form an integral part of the diet of many freshwater fish. It is therefore important to understand the effects that weed control practices have on this community in canal fishery watercourses. The principal forms of weed control operated in the Grand and Royal Canals include mechanical cutting, using a variety of boat-mounted and land-based apparatus, and chemical treatment using dichlobenil. The community composition and relative abundance of macroinvertebrates in control, mechanically cut and dichlobenil treated canal sites was recorded on three to five occasions between 1993 and 1994. The results indicated that Asellus aquaticus was the dominant organism at all canal locations. The land-based Mowing Bucket effected the greatest reduction in macroinvertebrate numbers in the immediate aftermath of the cut. This reflects the capacity of the machine to cut vegetation to canal bed level, thereby removing any substrate for colonisation. At all eight sites examined, macroinvertebrate numbers increased relatively rapidly following treatment and no adverse effect on dependent fish life resulted. The Office of Public Works policy of removing obstructive vegetation from a central navigation channel, while preserving weeded marginal fringes, minimises the impact of weed control operations on the macroinvertebrate fauna.     

Phenotypic plasticity of Capsella bursa-pastoris (Brassicaceae) and its effect on fitness in response to temperature and soil moisture

Capsella bursa-pastoris is one of the most common plants on earth. Although phenotypic plasticity of ecologically important traits possibly contributes to its wide geographic range, little is known about the plasticity of C. bursa-pastoris and its effects on its fitness. In a laboratory, we assessed the phenotypic plasticity in response to two representative climatic conditions: temperature and soil moisture. In addition, we quantitatively evaluated the relationship between phenotypic plasticity and fruit production. Most measured morphological and physiological traits exhibited plastic responses to temperature and fitness based on fruit production was maintained across temperatures. In contrast, no plasticity to soil moisture was detected, and plants produced fewer fruits in dry soil. Selection analysis revealed that the plasticity of the flowering time and the water-use efficiency had positive effects on fruit production over the tested temperature regime. These experimental results suggest that phenotypic plasticity probably enables C. bursa-pastoris to cope with heterogeneous temperature environments and thereby probably contributes to its wide geographic range.     

The interaction between Plectosporium alismatis and sublethal doses of bensulfuron-methyl reduces the growth of starfruit (Damasonium minus) in rice

Plectosporium alismatis is a fungal pathogen that is being investigated as a biocontrol agent for suppressing starfruit (Damasonium minus), a significant weed of Australian rice fields. The aim of this research was to study the effect of the fungus on weed competition and its interactions with chemical herbicides. Conidial germination was significantly reduced by Londax® (bensulfuron-methyl), while MCPA had no effects. Glasshouse trials showed evidence of synergism between the fungus and 1.56% of the recommended dose of Londax®, in reducing the weed growth. The application of inoculum (conidia suspended in water) in the glasshouse eliminated weed competition with rice. In the field, the reduction in weed growth caused by the fungus did not significantly eliminate starfruit competition with rice. This is thought to be due to the presence of other weeds.     

Effect of dew temperature,post-inoculation condition,and pathogen dose on suppression of scentless chamomile by Colletotrichum truncatum

Scentless chamomile, a noxious weed in western Canada, has a high natural tolerance to many herbicides. Colletotrichum truncatum, a host specific fungal pathogen, is suppressive to scentless chamomile when applied inundatively. A broadcast application was used at 200 L ha^?1 to evaluate biocontrol potential of this pathogen under a range of dew temperatures (DT), post-inoculation conditions (PIC), and pathogen doses (PD). A DT between 20 and 25°C was more conducive to infection, resulting in higher levels of disease and weed suppression as compared to 15 and 30°C. Under similar post-inoculation temperature regimes, disease was only slightly more severe in growth chambers than in the greenhouse. There was a positive linear relationship between the PD and weed suppression. An inoculum concentration >1×10⁸ spores mL^?1 reduced plant fresh weight by approximately 50% when compared to untreated controls. These results indicate that biocontrol of scentless chamomile using C. truncatum in the semi-arid Canadian prairies will likely encounter frequent non-favourable field conditions.     

A note on the role of Kachuga tectum gray (Reptilia: Chelonia) in controlling aquatic weeds

The herbivorous turtle Kachuga tectum consumes on wide variety of aquatic weeds and has potential as a biological agent against obnoxious weed growths.     

Effects of weed control on species composition of aquatic plants and bank plants and macrofauna in ditches

For several years, the effect of ditch management measures on the species composition of aquatic and bank flora and macrofauna were studied in a polder with a peat soil. The mowing basket increased the presence of floating plants whereas the ditch-scoop increased filamentous algae and submerged plant species. Monospecific stands of aquatic plants had fewer macrofaunal species than stands with many plant species. Direct and indirect effects of management measures on the macrofauna composition were shown.     

Pilot-scale process sensitivity studies for the scaleup of a fungal fermentation for the production of pneumocandins

The filamentous fungus Glarea lozoyensis produces a novel, pharmaceutically important pneumocandin (B(0)) that is used to synthesize a lipopeptide which demonstrates cidal activity against clinically relevant pathogens. A range of unwanted pneumocandin analogs are also produced by the organism. To maintain the unwanted impurities to acceptable levels upon scaleup, a good understanding of the impact of chemical and physical environment on the cell physiology is required, which benefits downstream processing. Pilot-scale studies were performed to determine the impact of dissolved oxygen, temperature, pH, and carbon dioxide on the process. Experiments included multiple fermenters (up to seven) at 0.07 and 0.8 m(3) scale using single source medium sterilization and inoculum. Gas blending was used to separate effects of dissolved oxygen from agitation. The process was significantly influenced by dissolved oxygen level. The critical dissolved oxygen tension (C(crit)) for growth was below 2% air saturation. The C(crit) for production of pneumocandin B(0) was 20% air saturation, with a significant reduction of the specific production rate below this value. In contrast, low dissolved oxygen levels produced a substantial increase of pneumocandins B(1), B(5), and E(0), while high dissolved oxygen levels produced a disproportionate increase of D(5). This sensivity to dissolved oxygen was independent of agitation within a power range of 2-15 kW/m(3). Broth viscosity was impacted below 10% dissolved oxygen, suggesting an effect on morphology. The process was shown to be sensitive to temperature but relatively insensitive to pH and carbon dioxide (in the exhaust gas) within the ranges studied. This scaledown analysis explained phenomena seen at pilot scale and helped define operating boundary conditions for successful scale up to 19 m(3).     

The impact of random frequency-dependent mutations on the average population fitness

ABSTRACT: BACKGROUND: In addition to selection, the process of evolution is accompanied by stochastic effects, such as changing environmental conditions, genetic drift and mutations. Commonly it is believed that without genetic drift, advantageous mutations quickly fixate in a halpoid population due to strong selection and lead to a continuous increase of the average fitness. This conclusion is based on the assumption of constant fitness. However, for frequency dependent fitness, where the fitness of an individual depends on the interactions with other individuals in the population, this does not hold. RESULTS: We propose a mathematical model that allows to understand the consequences of random frequency dependent mutations on the dynamics of an infinite large population. The frequencies of different types change according to the replicator equations and the fitness of a mutant is random and frequency dependent. To capture the interactions of different types, we employ a payoff matrix of variable size and thus are able to accommodate an arbitrary number of mutations. We assume that at most one mutant type arises at a time. The payoff entries to describe the mutant type are random variables obeying a probability distribution which is related to the fitness of the parent type. CONCLUSIONS: We show that a random mutant can decrease the average fitness under frequency dependent selection, based on analytical results for two types, and on simulations for n types. Interestingly, in the case of at most two types the probabilities to increase or decrease the average fitness are independent of the concrete probability density function. Instead, they only depend on the probability that the payoff entries of the mutant are larger than the payoff entries of the parent type.     

Production of microsclerotia of the fungal entomopathogen Lecanicillium lecanii (Hypocreales: Cordycipitaceae) as a biological control agent against soil-dwelling stages of Frankliniella occidentalis (Thysanoptera: Thripidae)

Abstract

Microsclerotia (MS) production by two isolates of Lecanicillium lecanii on various culture media is described, and the efficacy of MS against western flower thrips is evaluated. High concentrations of MS (2.9–3.1×10⁵·mL^?1) were produced in media with C:N ratios of 7.4:1 and 10.3:1 by isolate SN21. Bioassays using soil-incorporated MS resulted in significant infection and mortality of thrips.