Screening of Argentine native fungal strains for biocontrol of the grasshopper Tropidacris collaris: relationship between fungal pathogenicity and chitinolytic enzyme activity.

Tropidacris collaris (Orthoptera: Romaleidae) is a large and voracious grasshopper, which, in recent years, has become a recurrent pest in increasingly extensive areas of Argentina's northern provinces. In the present work chitinase activity was measured in 59 entomopathogenic fungal isolates native to Argentina, and the relationship between enzymatic activity and fungal virulence was assessed. Isolate LPSC 1067 caused the highest mortality on T. collaris nymphs (97.7 ± 1.22%). Nine isolates caused no mortality, while the remaining 49 caused mortalities ranging from 6.6 ± 0.3% (LPSC 770) to 91.06 ± 1.51% (LPSC 906). Several isolates revealed chitinolytic capabilities on test plates, although the activities differed with respect to the ratio of the chitin-decay-halo and fungal-colony diameters. A principal component analysis indicated that isolate LPSC 1067, obtained from a long-horned grasshopper (Orthoptera: Tettigoniidae), would be a potential candidate for T. collaris biocontrol because the strain exhibited the highest mortality, a shorter median lethal time, and a high enzymatic activity and growth rate.

First record of Fusarium verticillioides as an entomopathogenic fungus of grasshoppers.

Fusarium verticillioides (Saccardo) Nirenberg (Ascomycota: Hypocreales) is the most common fungus reported on infected corn kernels and vegetative tissues, but has not yet been documented as being entomopathogenic for grasshoppers. Grasshoppers and locusts represent a large group of insects that cause economic damage to forage and crops. Tropidacris collaris (Stoll) (Orthoptera: Acridoidea: Romaleidae) is a large and voracious grasshopper that in recent years has become an increasingly recurrent and widespread pest in progressively more greatly extended areas of some of in Argentina's northern provinces, with chemical insecticides being currently the only means of control. During February and March of 2008-09, nymphs and adults of T. collaris were collected with sweep nets in dense woodland vegetation at a site near Tres Estacas in western Chaco Province, Argentina, and kept in screened cages. F. verticillioides was isolated from insects that died within 10 days and was cultured in PGA medium. Pathogenicity tests were conducted and positive results recorded. Using traditional and molecular-biological methods, an isolate of F. verticillioides was obtained from T. collaris, and its pathogenecity in the laboratory was shown against another harmful grasshopper, Ronderosia bergi (Stål) (Acridoidea: Acrididae: Melanoplinae). The mortality caused by F. verticillioides on R. bergi reached 58 ± 6.53% by 10 days after inoculation. This is the first record of natural infection caused by F. verticillioides in grasshoppers.

Effect of phenanthrene and Rhodotorula glutinis on arbuscular mycorrhizal fungus colonization of maize roots.

The effect of the polycyclic aromatic hydrocarbon (PAH) phenanthrene and the yeast Rhodotorula glutinis on the arbuscular mycorrhizal fungus (AMF) Glomus geosporum colonizing maize roots, was studied. During a 90-day experiment, the highest G. geosporum colonization values were found in control plants. Mycorrhiza root length, measured both on the basis of percentage of root colonization and on the activity of succinate dehydrogenase, showed similar patterns in different phenanthrene treatments. The presence of phenanthrene in the substrate reduced G. geosporum intraradical colonization. The presence of R. glutinis did not enhance AMF colonization in the presence of phenanthrene. The biomass of the external mycelium estimated on the basis of the fatty acid 16:1 omega 5 concentration showed a progressive increase through time, and the amounts of this fatty acid differed among treated and untreated substrates. However, this increase was found to be lowest in the phenanthrene and Rhodotorula treatment at 60 days. There was less phenanthrene accumulation in roots of maize inoculated with AMF and the yeast than in roots inoculated only with AMF. A similar pattern was observed in the phenanthrene content of G. geosporum spores collected after 90 days.

Fluorescein diacetate hydrolysis as a measure of fungal biomass in soil.

The fatty acid methyl esters of lipids extracted from an agricultural soil in the preharvest period of soybean or middle growth cycle from wheat were characterized and quantified by gas-liquid chromatography. The fatty acids 18:2omega6 and 16:1omega5 were used as markers of saprotrophic and arbuscular mycorrhizal fungi. In parallel, biomass estimation through plate counts in selective media for cellulolytic and saprotrophic fungi was also performed all throughout a soybean crop or middle growth cycle of wheat. As an enzymatic method, the fluorescein diacetate (FDA) hydrolytic activity of the samples was determined. Owing to the high relationship exhibited by FDA hydrolysis with organic carbon and total nitrogen content of soil, the enzymatic activity was correlated with the microbial biomass estimated through marker lipids or plate counts. The results obtained point out that FDA hydrolysis may be used as a rapid, cheap, and reliable estimator of fungal biomass.

Extracellular ABTS-oxidizing activity of autochthonous fungal strains from Argentina in solid medium.

The screening for extracellular oxidases and peroxidases from autochthonous filamentous fungi isolated from different substrates is an important step towards the detection of extracellular fungal oxidative systems. Thirty-one autochthonous fungal strains from Argentina, belonging to different ecophysiological and taxonomic groups, were plate-screened for their ability to produce extracellular oxidoreductases. Modified Kirk solid medium containing the chromogen 2,2-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) was used to determine the presence of this extracellular activity. The fungi tested were grouped according to the colour intensity of the modified Kirk medium in: a) species without extracellular ABTS-oxidizing activity; b) species with low extracellular ABTS-oxidizing activity; c) species with moderate extracellular ABTS-oxidizing activity; d) species with high extracellular ABTS-oxidizing activity. The assay revealed extracellular ABTS-oxidizing activity in 90% of the strains tested. All species of Basidiomycetes used exhibited ABTS-oxidizing activity, except Laeticorticium roseum. Aspergillus terreus and Epicoccum purpurascens (Deuteromycetes) did not show extracellular oxidative activity on ABTS. Agrocybe aegerita, Amauroderma boleticeum, Cladosporium cladosporioides, Coriolopsis rigida, Grammothele subargentea, Graphium putredinis, Hexagona hydnoides, Hexagona papyraceae, Loweporus lividus, Peniophora albobadia, Phellinus everhartii, Phellinus gilvus; Phellinus linteus; Pleurotus laciniatocrenatus, Pycnoporus sanguineus, Rigidoporus ulmarius, Steccherinum rawakense, Talaromyces helicus, Trametes elegans, Trametes pavonia, Trametes villosa and Trichaptum sector are reported here for the first time as species capable of producing ABTS-oxidizing extracellular oxidorreductases.

Micobiota de la rizosfera y rizoplano de suelos contaminados con hidrocarburos / Mycobiota from rhizosphere and rhizoplane in hidrocarbon polluted soils

Se estudió la micobiota saprótrofa de suelo rizosférico y de rizoplano, de plantas que crecen en áreas contaminadas con hidrocarburos y en áreas control. Se aislaron e identificaron 52 especies fúngicas, de las cuales 39 pertenecen a los deuteromycetes, 3 a los ascomycetes, 8 a los zygomycetes (orden mucorales) y 2 son micelios estériles. Mediante el uso de métodos de ordenación, utilizándose el análisis de componentes principales, pudo diferenciarse las muestras obtenidas de áreas contaminadas de las no contaminadas. A pesar que las especies son las mismas en ambas áreas, sus frecuencias relativas permiten una discriminación de los muestreos