Influence of culture conditions on mycelial growth and bioluminescence of Gerronema viridilucens

Herein we describe a procedure for measuring the total light emission of the naturally bioluminescent tropical fungus Gerronema viridilucens and the optimization of culture conditions using multivariate factorial ANOVA. Cultures growing on an agar surface in 35 mm Petri dishes at 90% humidity show optimal bioluminescence emission at 25 degrees C in the presence of 1.0% sugar cane molasses, 0.10% yeast extract and pH 6.0 (nonbuffered). Temperature and pH are the most important factors for both mycelial growth and bioluminescence.

Germination and mycelial growth of Bipolaris euphorbiae Muchovej & Carvalho as influenced by herbicides and surfactants

Bipolaris euphorbiae Muchovej & Carvalho é um forte candidato para o controle de Euphorbia heterophylla L. (amendoim bravo). Este fungo pode ser aplicado em combinação com herbicidas para controlar um maior espectro de espécies daninhas. Para tanto, experimentos laboratoriais foram realizados para verificar a possibilidade da utilização de mistura de tanque de esporos de B. euphorbiae e herbicidas ou surfatantes recomendados para a cultura da soja. Crescimento micelial e germinação de conídios foram avaliados em meio BDA acrescido dos herbicidas, nas concentrações recomendadas dos produtos comerciais, oxasulfuron (80 g/ha), glifosato (4 L/ha), bentazon (1.5 L/ha), fomesafen (1 L/ha), chlorimuron-ethyl (80 g/ha), lactofen (1 L/ha) e imazetaphyr (1 L/ha) e dos surfatantes Energic (2 ml/L), Aterbane (2,5 ml/L), Silwet L-77Ag (1 ml/L), Herbitensil (2 ml/L) e Natur L'óleo (10 ml/L). Diluições dos herbicidas de 50% e 25% foram avaliadas com um consumo de calda equivalente a 300 L/ha. Os surfatantes foram somente utilizados nas concentrações recomendadas. O crescimento micelial não foi afetado por bentazon e fomesafen e apenas levemente por oxasulfuron. Porém, glifosato, chlorimuron-ethyl, lactofen, Energic, Herbitensil, Silwet, e Aterbane o reduziram drasticamente. A redução observada com imazetaphyr foi intermediária e Natur L' óleo promoveu o crescimento micelial. Na presença dos surfatantes, observou-se que todos permitiram uma porcentagem de germinação equivalente àquela alcançada na presença de água. Energic e Herbitensil causaram um retardamento expressivo. Com Herbitensil, o processo germinativo iniciou somente aos 120 minutos. Com herbicidas, foi observado que somente na presença de glifosato e imazetaphyr a germinação dos conídios não seguiu a tendência observada com água, como ocorreu com os outros produtos testados.

Isolation and mycelial growth of Diehliomyces microsporus: effect of culture medium and incubation temperature

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mycelial growth and microscopic characteristics of Thermomyces lanuginosus Tsiklinskaya in YpSs medium with different carbon sources

Muitos fungos termofílicos são conhecidos como sendo produtores de lipases e o potencial dessas enzimas nas reações de esterificação (reação inversa à hidrólise), há muito tem sido reconhecida. O aspecto do micélio e diversas observações microscópicas foram efetuadas em Thermomyces lanuginosus cultivado em Placas de Petri com meios de culturas YpSs (extrato de levedura + amido + agar) e mais quatro variações na fonte de carbono neste meio. As matérias-primas usadas e avaliadas foram pentaeritritol, ácido oléico e dioleato de pentaeritritol como possíveis fontes de carbono disponíveis ao microrganismo são comumente encontradas na indústria química como intermediários clássicos na síntese de diversos ésteres. Com o crescimento de Thermomyces lanuginosus em todos os meios propostos e a manutenção da capacidade de reprodução mostramos ser esse fungo altamente promissor em futuros trabalhos de biocatálise com microrganismos vivos.

Mycelial growth of two Lentinula edodes strains in culture media prepared with sawdust extracts from seven eucalyptus species and three eucalyptus clones

The in vitro mycelial growth of Lentinula edodes strains LE-95/01 and LE-96/18 were evaluated in solid culture media prepared with sawdust extracts from seven eucalyptus species (E. saligna, E. grandis, E. urophylla, E. pellita, E. paniculata, E. citriodora, and E. camaldulensis) and three eucalyptus clones (E. grandis × E. urophylla hybrids). Evaluations were made every 48 hours by means of colony diameter measurements (mean of four transversely-oriented measurements), during ten days of incubation in the dark at 25ºC ±1°C. The experimental design consisted of randomized blocks, and treatment means were compared by Tukey test. The culture medium prepared from E. citriodora sawdust extract was the most promising to grow L. edodes strains LE-96/18 and LE-95/01. L. edodes strain LE-96/18 presented the fastest mycelial growth after incubation for ten days, regardless of sawdust extract type used in the culture medium.

Comparative growth and pathogenicity of geographical isolates of Sclerotinia sclerotiorum on lentil genotypes

Isolates of Sclerotinia sclerotiorum were collected from infected lentil plants from 2 agro-ecological zones of Syria and used to study their comparative growth on culture media and pathogenicity on different lentil genotypes. The growth studies were carried out on Potato Dextrose Agar (PDA) growth media under laboratory conditions. Mycelial radial growth and sclerotial production were the parameters used to compare the isolates. Pathogenicity studies were carried out with selected isolates on 10 lentil genotypes, infected as detached shoots and as whole potted-plants in the plastic house. The isolates showed considerable variation in cultural characteristics through mycelial growth, mycelial pigmentation and sclerotial production in the media plates. There were significant differences in the growth and sclerotial production of most of the isolates, but no apparent correlation between mycelial growth and sclerotial production among the isolates. Genotype by isolate interactions was significant for the isolates tested for pathogenicity. These interactions, however, appeared to be caused by differences in virulence of the isolates and did not suggest the occurrence of distinct pathogenic races of the pathogen isolates.

Growth of and trehalase activity in the thermophilic fungusThermomyces lanuginosus

The thermophilic fungus,Thermomyces lanuginosus, was grown in a glucose-asparagine liquid medium. Optimal mycelial growth occurred at 50°C. The conditions for sporulation were different from those required for vegetative growth. the former being favoured by lower nitrogen level and temperature. Trehalase (α, α-glu coside-l-glucohydrolase, EC 3.2.1.28) was one of the most active glycosidases at 50°C. Non-sporulating mycelium had higher levels of this enzyme than the sporulating mycelium. Trehalase was synthesized constitutively and its activity appears to be controlled by catabolite repression.

In-vitro effect of fungicides on hyphal growth and sporogenesis of Lagenidium sp. isolated from Penaeus monodon larvae and Scylla serrata eggs

The sensitivity of Lagenidium, isolated from Penaeus monodon, Scylla serrata , to 34 antimycotics was determined. Effects on the development of vesicles, zoospores and mycelial growth were evaluated. Although mycoidal levels of the chemicals tested will be ideal for lethal treatment on control of the fungus, the high dose required may be lethal to the host, thus the use of mycostatic concentrations is more practical. Treatments of rearing water containing larvae, adult shrimps or crabs should be done only after preliminary tolerance experiments using at least the mycostatic dose prove to be safe for the hosts. Mycocidal doses can be used for determining disinfection doses of equipment and facilities used in rearing procedures as well as for destroying batches of infected larvae.

MYCELIAL DEVELOPMENT of Lentinula edodes IN CULTURE MEDIA WITH DIFFERENT ORGANIC SUBSTRATES

The radial mycelial growth of Lentinula edodes (Berk) Pegler, strain LE-96/13, was studied in culture media prepared with organic residues extract, by using substrates prepared with pineapple (Ananas comosus (L.) Merril) crown, Astrocaryum aculeatum Meyer peel, Theobroma grandiflorum Schum shell, Musa sp. (genomic group AAB, subgroup Pacovan) peel, and Musa sp. (genomic group AAB, subgroup Praia) peel, with three supplementation levels with wheat bran (0, 10 and 20%), and incubated at 25 degrees C. The experimental design was totally randomized, in a 5x3 factorial scheme, adding up 15 treatments with 4 repetitions, and each repetition corresponding to a Petri dish. The diameter of the colony was evaluated daily during nine days of incubation. After that period, it was verified that the highest mycelial growth averages of strain LE-96/13 of L. edodes were found in culture media prepared with T. grandiflorum Schum shell (whose supplementation with wheat bran was favorable for Mushroom development) and A. aculeatum Meyer peel (whose supplementation did not favor the mycelial growth of L. edodes in relation to the medium not supplemented).

Control of amylase production and growth characteristics of Aspergillus ochraceus

The growth and the extracellular amylase production by Aspergillus ochraceus were studied in a stationary culture medium. Maximum growth rate of this fungus was found after 5 days of incubation at 30° C, but maximum amylase production was obtained after 2 days. The highest amylase production were attained with lactose, maltose, xylose and starch as carbon sources. The extracellular amylase production and mycelial growth were influenced by the concentration of starch. Other carbohydrates supported growth but did not induce amylase synthesis and glucose repressed it, indicating catabolite repression in this microorganism. The presence of both mechanisms of induction and repression suggests that at least these multiple forms of regulation are present in A. ochraceus. Of the nitrogen sources tested, casaminoacids, ammonium nitrate and sodium nitrate stimulated the highest yield of amylase. Optimal amylase production was obtained at pH 5.0, but enzyme activity was found only in the 4.0-6.0 pH range. These results were probably due to the inhibitory effect of NH 4 +-N in the culture medium.

Growth of Pleurotus ostreatus in culture media based on formulated straw or grass

Mycelial growth strains of POS 98/38, POS 09/100, POS 09/101, and POS 09/102 of Pleurotus ostreatus were evaluated in culture media with various compositions based on extracts of substrates formulated with sugar cane bagasse together with straws and grasses, and with or without nitrogen supplementation. The evaluation was performed during incubation regularly with a ruler graduated in millimeters until total colonization of the culture medium contained in Petri dishes. The statistical model explaining the kinetics of mycelial growth of mushroom strains of P. ostreatus as a deterministic component has an exponential Gompertz function. The results show that the culture medium with sugar cane straw and brizantha grass (supplemented) showed the highest rates of mycelial growth, regardless of strain used compared to wheat straw-based culture media with had the lowest velocities of growth, regardless of supplementation and strains studied.

The interactive effects of temperature and osmotic potential on the growth of marines isolates of Fusarium solani

The mycelial growth of 18 Fusarium solani strains isolated from sea beds of the south-eastern coast of Spain was tested on potato-dextrose agar adjusted to different osmotic potentials with either KCl or NACl (-1.50 to -144.54 bars) in 10ºC intervals ranging from 15 to 35ºC. Fungal growth was determined by measuring colony diameter after 4 days incubation. Mycelial growth was maximal at 25ºC. The quantity and frequency pattern of mycelial growth of F. solani differ significantly at 15 and 25ºC, with maximal occurring at the highest water potential tested (-1.50 bars); and at 35ºC, with a maximal mycelial growth at -13.79 bars. The effect of water potential was independent of salt composition. The general growth pattern of F. solani showed declining growth at potentials below -41.79 bars. Fungal growth at 35ºC was always higher than that growth at 15ºC, of all the water potentials tested. Significant differences observed in the response of mycelia to water potential and temperature as main and interactive effects. The viability of cultures was increasingly inhibited as the water potential dropped, but some growth was still observed at -99.56 bars. These findings could indicate that marine strains of F. solani have a physiological mechanism that permits survival in environments with low water potential. The observed differences in viability and the magnitude growth could indicate that the biological factors governing potential and actual growth are affected by osmotic potential in different ways.

The Interactive Effects of Temperature and Osmotic Potential on the Growth of Aquatic Isolates of Fusarium culmorum.

The mycelial growth of 10 Fusarium culmorum strains isolated from water of the Andarax riverbed in the provinces of Granada and Almeria in southeastern Spain was tested on potato-dextroseagar adjusted to different osmotic potentials with either KCl or NaCl (−1.50 to−144.54 bars) at 10◦C intervals ranging from15◦ to 35◦C. Fungal growth was determined by measuring colony diameter after 4 d of incubation. Mycelial growth was maximal at 25◦C. The quantity and capacity of mycelial growth of F. culmorum were similar at 15 and 25◦C, with maximal growth occurring at −13.79 bars water potential and a lack of growth at 35◦C. The effect of water potential was independent of salt composition. The general growth pattern of Fusarium culmorum growth declined at potentials below −13.79 bars. Fungal growth at 25◦C was always greater than growth at 15◦C, at all of the water potentials tested. Significant differences were observed in the response ofmycelia to water potential and temperature as main and interactive effects. The number of isolates that showed growth was increasingly inhibited as the water potential dropped, but some growth was still observable at −99.56 bars. These findings could indicate that F. culmorum strains isolated from water have a physiological mechanism that permits survival in environments with low water potential. Propagules of Fusarium culmorum are transported long distances by river water, which could explain the severity of diseases caused by F.culmorum on cereal plants irrigated with river water and its interaction under hydric stress ormoderate soil salinity. The observed differences in growth magnitude and capacity could indicate that the biological factors governing potential and actual growth are affected by osmotic potential in different ways.

Tan spot: A new disease of pyrethrum caused by Microsphaeropsis tanaceti sp. nov

The isolation frequency of Microsphaeropsis sp. in spring in association with necrotic lesions on leaves in Tasmanian pyrethrum (Tanacetum cinerariifolium) fields has increased substantially since first identification in 2001. Examination of morphological features and sequencing of the internal transcribed spacer region (ITS) resulted in the identification of a new species, herein described as Microsphaeropsis tanaceti sp. nov. The pathogenicity of three M. tanaceti isolates to two pyrethrum cultivars was confirmed by inoculating glasshouse-grown plants in three experiments. No significant differences in the susceptibility of the two cultivars to infection by M. tanaceti were found. Symptoms were tan-coloured spots which coalesced around the margins of the leaves. Therefore, the name 'tan spot' is proposed for this new disease of pyrethrum. The sensitivity of seven M. tanaceti isolates to difenoconazole and azoxystrobin, commonly used fungicides for the management of foliar diseases in spring, was assessed under in vitro conditions. Sensitivity testing for difenoconazole was conducted using a mycelial growth assay on potato dextrose agar, whilst testing for sensitivity to azoxystrobin used a conidial germination assay on water agar. Microsphaeropsis tanaceti was found to be more sensitive to azoxystrobin than difenoconazole, with complete inhibition of conidial germination at concentrations above 0.625 µg a.i. mL-1. By comparison, concentrations of 50 µg a.i. difenoconazole mL-1 or greater were required for significant inhibition of mycelial growth. It therefore appears likely that there is currently some control of tan spot as a result of the use of azoxystrobin and to a lesser extent, difenoconazole, for the control of other diseases.