Mode of Infection of Ascochyta Blight of Chickpea Caused by Ascochyta rabiei

Process of infection and histological changes with Ascochyta blight of chickpea caused by A. rabiei (Pass.) Labr. were studied by light microscopy. Germ tubes from conidia of the fungus penetrate the stem tissue at the juncture of two epidermal cells and form subepidermal aggregates until the fourth day. On the sixth day, yellowing and necrotisation of host tissue coincides with formation of mature pycnidia. Fungus causes extensive damage to cellulosic cell walls of parenchymatous cortical and pith tissues in advance of invading hyphae indicating involvement of cell wall degrading enzymes. Lignified tissues, particularly xylem tracheary elements, remain intact.     

Formation of ascochitine by plant pathogens of the genus Ascochyta

Strains of fungi isolated from pulse crops: pea (Pisum sativum) and faba beans (Vicia faba) plants with symptoms of Ascochyta blight, footrot and stems lesions have been examined under laboratory conditions for their ability to produce ascochitine and metabolites toxic toArtemia salina. BothAscochyta pisi Lib and Ascochyta fabae LK Jones isolates formed ascochitine in yields of 20–480mg/kg. The highest yield of ascochitine was produced on rice and the lowest on maize grain. Ascochyta pinodes andPhoma medicaginis varpinodella (LK Jones) Boerema (formerlyAscochyta pinodella LK Jones) did not produce ascochitine. Crystalline ascochitine was found to be of moderate toxicity toArtemia salina larvae (LC₅₀ = 85μg/cm³BSM*). Extracts ofPhoma medicaginis var,pinodella cultures were found to be highly toxic toArtemia salina.     

Taxonomical re-evaluation of Phoma-like soybean pathogenic fungi

Coelomycetous fungi classified in Ascochyta, Phoma, and Phyllosticta have been recorded from spots on leaves and pods of soybeans. Based on the Genealogical Concordance Phylogenetic Species Concept, the authors suggest the re-evaluation of the taxonomic status of Phoma sojicola (syn. = Ascochyta sojicola) and Phyllosticta sojicola. In spite of the former delimitation of Phoma sojicola based on small differences in morphological features, it has proved to be identical to Phoma pinodella. Similarly, it was also confirmed that Phyllosticta sojicola was identical to Phoma exigua var. exigua. The authors supply tools for identification of Phoma-like fungi by combined conventional and molecular methods. Protein-encoding genes (tef1 and β-tubulin) were successfully applied within the Phoma genus to infer phylogenetic relationships.     

Artificial culture,host infection and pycnidial development of Ascochyta sorghina Sacc

Mode of penetration and subsequent development of Ascochyta sorghina Saccardo in the leaf tissues of sorghum (Sorghum vulgare Pers.) were studied under field conditions. Penetration by A. sorghina was observed both directly through the epidermis and indirectly through the stomatal openings in the leaves. The pathogen produced intercellular mycelium, which later became intracellular after death of cells around the infection site through action of toxic metabolites produced by the pathogen. The pycnidia developed through the growth of a subcuticular hyphal crust.     

Étude de la pycnogenèse chezPhomopsis mali (Schultz etSacc.)Rob.

Light induces the formation of pycnidia inPhomopsis mali. The induction caused by light can be conserved in darkness. The size and quantity of pycnidia vary with the illuminance. Under low and high illuminances physiologically different pycnidia appear.Phomopsis mali cultivated in darkness in the presence of other microorganisms (fungi or bacteria) can fructify and produces pycnidia similar to those formed under low illuminance and in natural conditions. Our experiments indicate the presence of a pycnogenesis-inducing substance that can exists in different forms and induces the formation of the different pycnidia.     

Solar UV-B Radiation Inhibits the Growth of Antarctic Terrestrial Fungi

We tested the effects of solar radiation, and UV-B in particular, on the growth of Antarctic terrestrial fungi. The growth responses to solar radiation of five fungi, Geomyces pannorum, Phoma herbarum, Pythium sp., Verticillium sp., and Mortierella parvispora, each isolated from Antarctic terrestrial habitats, were examined on an agar medium in the natural Antarctic environment. A 3-h exposure to solar radiation of >287 nm reduced the hyphal extension rates of all species relative to controls kept in the dark. Pythium sp. cultures exposed to solar radiation for 1.5 h on five consecutive days were most sensitive to radiation of >287 nm, but radiation of >313 nm also inhibited growth to a lesser extent. Radiation of >400 nm had no effect on hyphal growth relative to controls kept in the dark. Short-wave solar UV-B radiation of between 287 and 305 nm inhibited the growth of Pythium sp. hyphae on and below the surface of the agar medium after 24 h, but radiation of ≥345 nm only reduced the growth of surface hyphae. Similar detrimental effects of UV-B on surface and, to a lesser extent, submerged hyphae of all five fungi were shown in the laboratory by using artificial UV-B from fluorescent lamps. A comparison of growth responses to solar radiation and temperature showed that the species that were most resistant to UV radiation grew fastest at higher temperatures. These data suggest that solar UV-B reduces the growth of fungi on the soil surface in the Antarctic terrestrial environment.     

Effect of food level on the growth and survival of laboratory-reared larvae of bay anchovy (anchoa mitchilli Valenciennes) and scaled sardine (harengula pensacolae Goode & Bean)

The effect of food levels on the growth and survival of laboratory-reared larvae of scaled sardines (Harengula pensacolae Goode & Bean) and bay anchovies (Anchoa mitchilli Valenciennes) was determined. Wild zooplankton, most of which was copepod nauplii and copepodids, was used as food. H. pensacolae larvae grew 0.80 mm/day at a low (444 organisms/1) and high food level (1324 organisms/1). Survival was appreciably better (14.5%) at the low than (3.5%) at the high food level, 23 days after hatching. A. mitcchilli larvae grew faster at medium and high food concentrations. Average growth rates for three replicated experiments were 0.48 mm/day at low food (621–692 organisms/1), 0.50 mm/day a t medium food (1330–1688 organisms/1), and 0.54 mm/day at high food (2811–3323 organisms/1). Survival of A. mitchilli larvae, 20 to 28 days after hatching, ranged from 0.0 to 17.8% and was better at medium and high food levels than at the low food level. The resultsng indicate that 500 to 1000 copepod nauplii and copepodids/1 provided adequate food for rearing H. pensacolae but 1500–2000/1 were consistently required to rear A. mitchilli.     

Notes on various plant-inhabiting fungi from Hachijo Island, Tokyo (1)

Among plant-inhabiting fungi collected in June 2001 and in September 2002 on Hachijo Island, Tokyo, four fungi are described in this article. They consist of two new species, namely Stagonospora hachijoensis on Miscanthus sinensis var. condensatus and Ascochyta ixorae on Ixora chinensis, and two fungi newly added to the Japanese mycoflora, namely Discosiella cylindrospora on Callistemon speciosum and Robillarda sessilis on Parthenocissus tricuspidatus.     

Fungal interaction between Trichoderma spp. and Pleurotus ostreatus on the enriched solid media with licorice Glycyrrhiza glabra root extract

The aim of this study is to investigate the antifungal activity of mycelia of Pleurotus ostreatus (white oyster mushroom) and licorice (Glycyrrhiza glabra) root extract against three undesirable fungi. They are Trichoderma spp., Trichoderma harzianum I and Trichoderma harzianum II which was tested on PSA (potato sucrose agar) medium enriched with licorice (Glycyrrhiza glabra) root extract (PSA-G media) using three concentrations (0.05, 0.10 and 0.20 g/L) in alone and dual cultures. Trichoderma spp. showed less mycelial growth of 8.75, 9.17 and 9.50 mm/day on PSA-G0.05, PSA-G0.1 and PSA-G0.2 respectively compared with 10.25 mm/day on fresh PSA (control) in dual culture. The best mycelial growth inhibition was recorded on PSA-G0.2 (14.97%) by T. harzianum II in alone culture opposite 63.72% in dual ones. The lower mycelial growth rate of T. harzianum I was 17.75 mm/day on PSA-G0.1 (0.10 g/L). In dual culture, overgrowth time of T. harzianum I had 5 days compared as approx. 6 days in alone culture. Generally, when the concentration of licorice extract increased, the mycelial growth rate of the undesirable fungi decreased. Also, all PSA-G media, especially PSA-G0.2, indicated low growth averages compared with the control (fresh PSA) against the pathogen while this concentration encourages growth of oyster mushroom. Also, this concentration reduced the density of sporulation of green molds; therefore, this concentration can be applied to reduce influence this pathogen in cultivation farm.     

Comparison of the laccases, molecular marker proteins, and induction of pycnidia by three species of botryosphaeriaceous fungi

Three species of botryosphaeriaceous fungi,Botryosphaeria sp. isolate MAMB-5,Botryosphaeria ribis andLasiodiplodia theobromae, were compared for the production of pycnidia and laccases. Laccases were produced both intra- and extra-cellularly when the fungi were cultivated on basal medium in the presence and absence of veratryl alcohol, withBotryosphaeria sp. MAMB-5 showing the highest enzyme titres. Electrophoretic examination of intracellular marker proteins (esterases and phosphatases) and laccases indicated that the three species were genetically distinctly different, although the laccase zymograms for the three fungi showed similarity. The production of pycnidia occurred under continuous lighting at 28°C, but conditions differed among the three fungal species. Production could be induced on artificial media (potato-dextrose and oat agar) under stress-induced conditions where the mycelium was stimulated by physical abrasion, and in the case ofBotryosphaeria sp. isolate MAMB-5 on eucalypt woodchips. Evidence is presented that veratryl alcohol facillitated the secretion of intracellular-localised laccases into the extracellular medium.     

Micropropagation of Araucaria excelsa R. Br. var. glauca Carrière from orthotropic stem explants

The objectives of the present work were in vitro propagation of Araucaria excelsa R. Br. var. glauca Carrière (Norfolk Island pine) with focus on the evaluation of the mean number of shoots per explant (MNS/E) and mean length of shoots per explants (MLS/E) produced by different parts of the orthotropic stem of A. excelsa R. Br. var. glauca in response to plant growth regulators. Norfolk Island pine axillary meristems responded very well to the 2-iso-pentenyl adenine (2iP) and thidiazuron (TDZ) levels. Explants taken from stem upper segments in the media containing 2iP had a higher MNS/E (3.47) and MLS/E (6.27 mm) in comparison to those taken from stem lower segments, which were 0.71 and 0.51 mm, respectively. Using 0.045 μM TDZ in the MS medium not only resulted in 4.60 MNS/E with 7.08 mm MLS/E but proliferated shoots showed a good performance as well. Investigating the best position of stem explant on mother plant as well as the best concentrations of growth regulators were performed which were useful for efficient micropropagation of this plant. Thirty three percent of explants were rooted in the MS medium containing 3 % sucrose, supplemented with 7.5 μM of both NAA and IBA for 2 weeks before transferring to a half strength MS medium without any growth regulator. Plantlets obtained were acclimatized and transferred to the greenhouse with less than 20 % mortality. This procedure considered the first successful report for regeneration and acclimatization of A. excelsa R. Br. var. glauca plantlet through main stem explants.     

The effect of a preparation from Chaetomium fungi on the growth of phytopathogenic fungi

We studied the fungicidal activity of a biological preparation from the fungi of the genus Chaetomium against soil phytopathogenic fungi Rhizoctonia solani and Fusarium oxysporum. The inhibitory effect of the preparation under study depended on its concentration, duration of storage, and growth characteristics of pure cultures of the phytopathogens. The highest (98.8%) inhibitory activity was observed on the third day of the interaction with Rhizoctonia solani. After a 2-year storage, this preparation was capable of inhibiting the growth of phytopathogens only at high doses. The preparation precluded the development of a bare patch but increased the productivity of potato plants. The preparation may serve as an alternative to chemical fungicides for plant protection.     

Response of selected microbial strains and their consortia to the presence of automobile paints: Biofilm growth,matrix protein content and hydrolytic enzyme activity

The goal of the current study was to examine the effects of pollutants (White color – CP; Metallic red color – FM; Thinner – CN; Thinner for rinsing paint – MF; Basic color (primer) – FH) originating from the automotive industry on the biofilm growth, matrix protein content, and activity of the hydrolytic enzymes of selected microbial strains in laboratory conditions that mimic the bioreactor conditions. The chosen microorganisms (bacteria, yeasts, and fungi) were isolated from automotive industry wastewater. Pure microbe cultures and their consortia were injected into AMB Media carriers and developed into biofilms. The use of AMB media carriers has been linked to an increase in the active surface area colonized by microorganisms. Afterwards, the carriers were transferred to Erlenmeyer flasks with nutrient media and pollutants at a concentration of 200 μL/mL. The current study found that, depending on the microbial strain, development phase, and chemical structure, the assessed pollutants had an inhibitory or stimulatory influence on the growth of single cultures and their consortia. Statistical analysis found positive correlations between the protein content in the matrix and the biofilm biomass of Rhodotorula mucilaginosa and consortia in CP and FH media, respectively. The proteolytic activity of Candida utilis was very pronounced in media with MF and CN. The best alkaline phosphatase activity (ALP) was achieved in the CN medium of R. mucilaginosa. Acid invertase activity was the highest in the FM and CP media of Escherichia coli and consortia, respectively, whereas the highest alkaline invertase activity was measured in the MF medium of E. coli. A positive correlation was confirmed between ALP and the biofilm biomass of R. mucilaginosa in CP and CN media, as well as between ALP and the biofilm biomass of Penicillium expansum in FM medium. The findings provide novel insights into the extracellular hydrolytic activity of the investigated microbial strains in the presence of auto paints, as well as a good platform for subsequent research into comprehensive biofilm profiling using modern methodologies.     

Isolation of an endophytic fungus producing baccatin III from Taxus wallichiana var. mairei

The objective of this study was to isolate endophytic fungi producing baccatin III from yew for the purpose of baccatin III and paclitaxel manufacture. Surface sterilized bark of Taxus wallichiana var. mairei was used as source material with potato dextrose agar culture medium for isolation of endophytic fungi. Fungal cultures were extracted with a mixture of chloroform/methanol (1:1, v/v) and the baccatin III in the extracts was determined and authenticated with LC–MS. An endophytic fungus that produced baccatin III was identified by ITS rDNA and 26S D1/D2 rDNA sequencing. A total of 192 endophytic fungal strains were isolated from T. wallichiana var. mairei. Only one of the 192 strains produced baccatin III and it was identified as Diaporthe phaseolorum. The productivity of this strain cultured in PDA culture medium was 0.219 mg/l. The isolated endophytic fungus produced baccatin III at a relatively high level and shows promise as a producing strain for baccatin III and paclitaxel manufacture after strain improvement.     

Association of mating-type with mycelium growth rate and genetic variability of Fusarium culmorum

Background

Barley is an important crop used widely in Europe for food production, feed and malting. Unfortunately it is often colonised by fungi from the Fusarium genus. Fusarium culmorum is a global pathogen causing root rot and crown rot in small-grain cereals, resulting in a reduction in yield and grain quality. F. culmorum produces the highly toxic chemicals trichothecenes. Experimental

Procedures

Chemotypes and mating-type idiomorphs (MAT) were identified using Polymerase Chain Reactions (PCR) and genetic diversity was determined using Sequence-related Amplified Polymorphism (SRAP) and Random Amplified Polymorphic DNA (RAPD). Physiological features such as mycelium growth rate were also evaluated.

Results

As many as 94% of isolates was classified as a 3ADON producing and only two isolates displayed NIV chemotype. The average growth rate at 15°C and 25°C equalled 5.32 mm/day and 13.5 mm/day, respectively. The MAT idiomorph amplification revealed that 60% of isolates possessed MAT1-2 idiomorph. Among 32 obtained SRAP and RAPD markers, eight were associated with mycelium growth rate.

Conclusions

It was shown first time that F. culmorum isolates with MAT1-2 idiomorph in the genome grew slower than these with MAT1-1. High level of genetic variability was determined based on amplification of SRAP and RAPD markers.     

New species of <Emphasis Type="Italic">Moristroma</Emphasis> (Ascomycetes) and phylogenetic position of the genus

The loculoascomycete Moristroma quercinum sp. nov. and Moristroma japonicum sp. nov. are described from Northern Europe (Denmark, Lithuania, Sweden) and Japan, respectively. M. quercinum is reported from wood of Quercus robur and Q. petraea, and M. japonicum is reported from wood of Quercus mongolica var. grossoserrata. Ascostromata of both species were found on hard heartwood of attached or shed branches. The two new species differ from the type species of the genus, M. polysporum, by the presence of pycnidia, and by the size of ascostromata, asci and ascospores. Drawings illustrate ascostromata, pycnidia, asci, hamathecium and ascospores of the two new species. A phylogenetic analysis suggests that Moristroma belongs to the Chaetothyriomycetes, rather than to the Dothideomycetes as previously suggested.     

Sporulation et mycosporines chez un champignon imparfait: Ascochyta fabae

Sporalation and mycosporins in the Deuteromycete Ascochyta fabae .
To investigate the sporogenic activity of mycosporins, a strain of Ascochyta fabae Speg., in which conidiogenesis and mycosporin synthesis are easily induced, has been selected. In darkness, few mature pycnidia and little mycosporin are produced. Dry weight and mycosporin production were followed during mycelial growth and the conidiogenic phase, in both illuminated and non-illuminated cultures: Mycosporin production appears closely linked with the morphogenic differentiation; its synthesis starts when the exponential growth is over. There is a good correlation between conidiogenesis and mycosporin production.
Irrespective of illumination, there is no increase in the rate of sporulation when maycosporins are added at different concentration levels. This cannot be explained by impermeability of the hyphae to mycosporins since uptake starts after a lag phase of 7 days and then proceeds until 15 to 20 days; at the same time the mycelium of A. fabae keeps its sporulation competence until 16 days after transfer to complete darkness. Mycosporin accumulated into conidiospores – 3 to 5% of dry weight – is not metabolized during the germination; on the contrary, a slight synthesis occurs after 20 h of germination.
Mycosporins thus seem to be secondary metabolites appearing as products from a type of metabolism that occurs at a very low level during mycelial growth and increases its activity during reproductive morphogenesis in numerous species of fungi.     

Agropyrenol and agropyrenal,phytotoxins from Ascochyta agropyrina var. nana,a fungal pathogen of Elitrigia repens

A strain of Ascochyta agropyrina var. nana, a fungal pathogen of the perennial weed Elytrigia repens, produced several toxins in a liquid medium, and its primary toxin, named agropyrenol, was characterized as a substituted salicylaldehyde on the basis of its chemical and spectroscopic properties. Its absolute stereochemistry was determined by Mosher’s method. Two other minor metabolites were isolated from the same culture and named agropyrenal and agropyrenone, respectively. They were characterized as a trisubstituted naphthalene carbaldehyde and a pentasubstituted 3H-benzofuranone, respectively, using the same techniques. When assayed on leaves of several weed plants, i.e., Mercurialis annua, Chenopodium album and Setaria viridis, agropyrenol proved to be phytotoxic, causing the appearance of necrotic lesions, agropyrenal was less active, while agropyrenone was inactive. None of the compounds showed antibiotic, fungicidal or zootoxic activity.     

Phoma sp. FOM-8108, a producer of gentisylquinones, isolated from sea sand

A fungal strain, FOM-8108, that was isolated from sea sand was found to produce gentisylquinone and chlorogentisylquinone, inhibitors of neutral sphingomyelinase. The fungus grew well under normal conditions, in the darkness, or on various agar media, but typical morphological characteristics were not observed to determine its taxonomy. Therefore, culture conditions were studied extensively, resulting in formation of a number of pycnidia by the fungus on the media containing seawater or on natural substrates such as hydrangea leaves, gardenia leaves, and rice straw under natural light or near-ultraviolet radiation exposure. Eventually, from the morphological characteristics of pycnidia, conidiogenous cells, and conidia, strain FOM-8108 was considered to belong to the genus Phoma. Culture studies showed seawater in the fermentation medium was necessary for the strain to produce gentisylquinones. Particularly, a full production of chlorogentisylquinone, which has a chloride ion in the structure, was performed in the medium with higher concentration (75%–100%) of seawater. Received: June 27, 2001 / Accepted: December 14, 2001     

Metabolic activity of <Emphasis Type="Italic">Aspergillus niger</Emphasis> and <Emphasis Type="Italic">Fusarium lateritium</Emphasis> induced by ethoxylated oleyl cetyl alcohol and their bioremediation and biotechnological potential

The effect of ethoxylated oleyl cetyl alcohol (Henkel, Serbia) on the growth and metabolic activity of Aspergillus niger and Fusarium lateritium was in the focus of this paper. The fungi were isolated from wastewater of Lepenica River (Kragujevac, Serbia) at a place where municipal wastewater discharged into the river. The fungi were grown in Czapek-Dox liquid nutrient medium without and with addition of 0.5% pollutant. The physico-chemical and biochemical changes of pH, total biomass dry weight, quantity of free and total organic acids, proteolytic activity and quality of carbohydrates were evaluated from 4-th to 19-th day of fungal growth. The capacity of fungi to decrease concentration of pollutant in medium was determined by cobalt thiocyanate method. The pollutant caused an inhibitory effect on biomass dry weight of A. niger and F. lateritium for 8.50 and 30.61%, respectively. Among tested fungi, A. niger had the better biodegradation capacity (83%) than F. lateritium (65%). Alkaline protease activity of A. niger enhanced in the presence of pollutant for 7.6% whereas the enzyme of F. lateritium retained about 62.2% activity. Overall, the obtained results indicate the potential application of tested fungi in wastewater treatment, detergent industry and biotechnology.