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As part of long term studies focused in the lichenized mycobiota diversity from southern Argentina, two species of the family Lobariaceae (Peltigerales), Pseudocyphellaria pluvialis and Sticta longipes, are registered for the first time in Argentina. These species were previously reported as endemic from southern Chile. Brief descriptions, illustrations and information about the distribution and ecology of the species are provided.

The most accepted method for staining arbuscular mycorrhiza (AM) in vascular plants has been one proposed by Phillips & Hayman in 1970. In particular, for the study of AM in bryophytes (s.l.) [Anthocerotophyta, Bryophyta (s.s.), Marchantiophyta] some authors have introduced modifications tothis technique. Even though all these protocols stain AM, their main disadvantage is related to the result of material maceration (e.g. over-softening or completely destroying plant cells due to the high temperaturesused, the high concentrations of reagents or the long-term exposure to aggressive chemicals). In order to optimise the results for the observation of AM in this group of plants, a modification is presented to the traditional staining technique. In the protocol here proposed, 70% ethanol is used as fixative and first clarifier, 1% potassium hydroxide (KOH) (80 °C, 20 min) as a second clarifier; 1% hydrochloric acid (HCl) (50 °C, 10 min) as an acidifier and 0.05% trypan blue (60 °C, 20 min) for dyeing. This improved protocol is not destructive, it is fast to perform and it is of wide application since it allows staining the AM in bryophytes. Fil: Cottet, Agustina Celeste. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina Fil: Scervino, Jose Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina Fil: Messuti, Maria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina

El fósforo (P) es un macronutriente poco biodisponible para las plantas y limitante para su crecimiento. El uso de hongos solubilizadores de P (HS) es una alternativa prometedora en el aprovechamiento de la fertilización fosforada. Sin embargo, aún no se ha establecido si los HS tienen efectos antagónicos con la flora benéfica del suelo, como los hongos micorrícico arbusculares (HMA). Este trabajo estudia el efecto de HS sobre el desarrollo micorrícico pre-infectivo, el primer paso en la colonización de la raíz. Se expusieron esporas del HMA Gigaspora rosea a distintas concentraciones de exudados (0; 0,01; 0,1; 1; 3%) de HS (Talaromyces helicus L, T. helicus N, T. diversus y Penicillium purpurogenum) en un sistema in vitro con medio semilíquido. Se evaluó la germinación, elongación, ramificación y morfología del micelio pre-infectivo. Se observó que T. diversus incrementó solo la germinación y que T. helicus L promovió el desarrollo pre-infectivo. Estos resultados demuestran que la germinación y el desarrollo del micelio pre-infectivo de G. rosea pueden ser modificados en longitud y ramificación en función de la cepa solubilizadora estudiada, sugiriendo que las interacciones microbianas podrían impactar positivamente sobre la nutrición fosforada vegetal mediante la modificación del estadio previo a la simbiosis micorrícica. Microbial interactions: effect of biocontrol and P solubilizing fungi in pre-symbiotic stages of arbuscular mycorrhizal fungi. Phosphorus (P) is an essential macronutrient with a low bioavailability in soils, thus limiting the plant growth. The use of P solubilizing fungi (HS) is a promising alternative to increase the efficiency of P-fertilizers. However, it is not fully understood if HS have antagonist effects on beneficial soil microbiota, such as arbuscular mycorrhizal fungi (HMA). This work studies the effect of HS with biocontrol abilities against phytopathogenic fungi on the pre-infective stages development of HMA, the first step in mycorrhizal root colonization. Spores of the HMA Gigaspora rosea were exposed in vitro to different exudate concentrations (0; 0,01; 0,1; 1; 3%) of different HS fungal strains (Talaromyces helicus strain L7B, T. helicus strain N24, T. diversus and Penicillium purpurogenum) in a system containing semi-liquid medium. Spores germination, elongation, ramification and morphology of the pre-infective mycelium of HMA were evaluated. We observed that only T. diversus increased germination while T. helicus L7B promoted the elongation, ramification and produced changes in the mycelial morphology without affecting spores’ germination. These results showed that germination and pre-infective development of G. rosea can be modified in length and ramification as a function of the identity of the HS strain used, suggesting that microbial interactions can positively impact on plant P nutrition through the modification of the previous stage to mycorrhizal symbiosis. Fil: Della Mónica, Ivana Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentina Fil: Godeas, Alicia Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Biodiversidad y Biología Experimental y Aplicada. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Biodiversidad y Biología Experimental y Aplicada; Argentina Fil: Scervino, Jose Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigaciones en Biodiversidad y Medioambiente. Universidad Nacional del Comahue. Centro Regional Universidad Bariloche. Instituto de Investigaciones en Biodiversidad y Medioambiente; Argentina

Phosphorus availability is a major limiting factor for yield of most crop species. The objective of this study was to compare the solubilization of three sources of phosphorus (P) by different fungal isolates and to determine the possible mechanisms involved in the process. Talaromyces flavus (S73), T. flavus var flavus (TM), Talaromyces helicus (L7b) and T. helicus (N24), Penicillium janthinellum (PJ), and Penicillium purpurogenum (POP), fungal strains isolated from the rhizosphere of crops, are known to be biocontrol agents against pathogenic fungi. The P solubilization efficiency of these fungal strains in liquid media supplemented either with tricalcium phosphate (Ca3(PO4)2; PC), aluminum phosphate (AlPO4; AP), or phosphorite (PP) depended on the source of P and the fungal species. The type and concentration of organic acids produced by each species varied according to the source of available P. In the medium supplemented with PC, the highest proportion was that of gluconic acid, whereas in the media supplemented with the other P sources, the highest proportion was that of citric and valeric acids. This suggests that the release of these organic compounds in the rhizosphere by these microorganisms may be important in the solubilization of various inorganic P compounds. Results also support the hypothesis that the simultaneous production of different organic acids by fungi may enhance their potential for solubilizing insoluble phosphate.

Exudates of a dark septate endophyte (DSE) identified as Dreschlera sp., a common endophyte isolated by the inner cortical cells of the grass Lolium multiflorum , were put in contact with the arbuscular mycorrhizal fungus (AMF) Gigaspora rosea . These exudates stimulated the hyphal length and the hyphal branching of the AMF. A negative effect on the extramatrical phase of the AMF was detected. This is the first report to show how exudates of DSE can affect the development of AMF. These results show that DSE could be modifying the mycorrhizal status of the plants, modulating a different symbiosis in the rhizosphere.

No clear data are available on how flavonoids from different chemical groups affect root colonization by arbuscular mycorrhizal fungi (AMF) and whether flavonoids affecting the presymbiotic growth of AMF also affect root colonization by AMF. In the present work, we compared the effect of flavones (chrysin and luteolin) and flavonols (kaempferol, morin, isorhamnetin, and rutin) on root colonization (number of entry points and degree of root colonization) of tomato plants ( Lycopersicum esculentum L.) with the effect of these flavonoids on the presymbiotic growth of these AMF, which has been reported in a recent study. With all tested AMF ( Gigaspora rosea , Gigaspora margarita , Glomus mosseae, and Glomus intraradices) a correlation between the number of entry points and the percentage of root colonization was found. When the number of entry points was high, root colonization was also enhanced. Application of the flavones chrysin and luteolin and of the flavonol morin increased the number of entry points and the degree of colonization,whereas the flavonols kaempferol, isorhamnetin, and rutin showed no effect. These results show that in contrast to their effect on the presymbiotic growth of the AMF on the level of root colonization, the tested flavonoids do not exhibit a genus- and species-specificity. Moreover, comparison of our data with the data obtained by J.M. Scervino, M.A. Ponce, R. Erra-Bassells, H. Vierheilig, J.A. Ocampo, and A. Godeas. (2005a. J. Plant Interact. 15: 22–30) indicates that a positive effect on the hyphal growth of AMF does not necessarily result in an enhanced AM root colonization, further indicating that the mode of action of flavonoids at the level of root colonization is more complex.

The effect of different concentrations (0.5, 2 and 8mM) of apigenin and its glycosidated form 5,7,4 0 -hydroxy flavone glycoside on arbuscular mycorrhizal (AM) fungal spore germination, hyphal growth, hyphal branching, the formation of entry points and root colonization of Gigaspora. rosea, Gi. margarita, Glomus mosseae and G. intraradices was tested. The lowest apigenin concentration (0.5mM) nearly doubled hyphal branching, the formation of entry points and root colonization of all four tested fungi, whereas higher concentrations (2 and 8mM) nearly doubled the hyphal growth of Gi. margarita, G. mosseae and G. intraradices. In none of the treatments with the apigenin-glycoside any effect on AM fungi could be observed. Our data show that apigenin exhibits an AM fungal genus and even species activity and we provide strong evidence that glycosidation results in a loss of its activity towards AM fungi. r 2006 Elsevier Ltd. All rights reserved.

The effect of flavonoids isolated from arbuscular mycorrhizal (AM) colonized and noncolonized clover roots on the number of entry points and percentage of root colonization of tomato (Lycopersicum esculentum L.) by Gigaspora rosea, Gi margarita, Glomus mosseae and G. intrarradices symbionts was determined. With fungi of both genera, a correlation between the number of entry points and the percentage of root colonization was found in the presence of some of the tested flavonoids. The flavonoids acacetin and rhamnetin, present in AM clover roots, inhibited the formation of AM penetration structures and the AM colonization of tomato roots, whereas the flavonoid 5,6,7,8,9-hydroxy chalcone, which could not be detected in AM clover root, inhibited both parameters. The flavonoid quercetin, which was present in AM clover roots, stimulated the penetration and root colonization of tomato by Gigaspora. However, the flavonoids 5,6,7,8-hydroxy-4'-methoxy flavone and 3,5,6,7,4'-hydroxy flavone, which was not found in AM clover root, increased the number of entry points and the AM colonization of tomato roots by Gigaspora. These results indicated that flavonoids could be imnplicated in the process of regulation of AM colonization in plant root, but its role is highly complex and depend not only on flavonoids, but also on AM fungal genus or even species.

The flavonoids 5,6,7,8,9-hydroxy chalcone, 3,7-hydroxy-4′methoxy flavone, 5,6,7,8-hydroxy-4′-methoxy flavone and 3,5,6,7,4′-hydroxy flavone can be detected only in non-mycorrhizal roots of white clover, but not in mycorrhizal roots, whereas the flavonoids acacetin, quercetin and rhamnetin are only present in mycorrhizal roots. We tested the effect of several concentrations of these compounds on spore germination, hyphal growth, hyphal branching, formation of clusters of auxiliary cells and of secondary spores of the arbuscular mycorrhizal fungi Gigaspora rosea, Gigaspora margarita, Glomus mosseae and Glomus intraradices. Our results indicate that depending on the flavonoid, the tested compounds are involved at different stages in the regulation of mycorrhization. This hypothesis is strengthened by their differing effect on several AM fungal growth parameters. Furthermore, our study provides more data on the AM fungus genus/species specificity of flavonoids.

White clover (Trifolium repens) plants were grown in the presence or absence of the arbuscular mycorrhizal fungus Glomus intraradices. Flavones, 4',5,6,7,8-pentahydroxy-3-methoxyflavone and 5,6,7,8-tetrahydroxy-3-methoxyflavone, as well as two flavones 3,7-dihydroxy-4'-methoxyflavone and 5,6,7,8-tetrahydroxy-4'-methoxyflavone never previously reported in plants, were isolated. The known 3,5,6,7,8-pentahydroxy-4'-methoxyflavone, 2',3',4',5',6'-pentahydroxy-chalcone, 6-hydroxykaempferol, 4',5,6,7,8-pentahydroxyflavone and 3,4'-dimethoxykaempferol were also obtained. Analysis of extracts obtained from roots and shoots revealed that the compositions of the flavonoid mixtures varied with growing conditions. Quercetin, acacetin and rhamnetin accumulated in roots of inoculated plants, whereas they were not detected in non-inoculated plants.

Exudates of Rhodotorula mucilaginosa, a yeast commonly found in the rhizosphere, increased hyphal length of the arbuscular mycorrhizal (AM) fungi Gigaspora rosea and Gigaspora margarita. Rhodotorulic acid (RA), a siderophore compound obtained from R. mucilaginosa exudates, increased hyphal length and branching. Thus, the increase in the number of entry points and the higher AM root colonization of tomato plants in the presence of RA can at least partially be explained by the positive effect of RA on the pre-symbiotic stages of the AM fungi.

We tested the effects of the flavonoid 3-methoxi-5,6,7,8-hydroxy-4'hydroxy flavone (NMHTV) isolated from shoots of non arbuscular mycorrhizal (AM) inoculated clover, and of the flavonoids 5,6,7,8-hydroxy-3-methoxy flavone (MH-1); 5,6,7,8-hydroxy-4'- hydroxy flavone (MH-2); and 5,7-hydroxy-3,4'-methoxy flavone (MH-3); isolated from AM clover (Trifolium repens) shoots, on spore germination, hyphal length, hyphal branches and the number of cluster of auxiliary cells or the number of secondary spores (Presymbiotic stage) and on the number of entry points and the percentage of AM colonized root of tomato (Lycopersicum esculentum) by the AM fungi Gigaspora rosea, Giaspora margarita, Glomus mosseae and Glomus intraradices (Symbiotic stage). Non significant effects of the flavonoids isolated from the shoot of mycorrhizal colonized clover on the presymbiotic and symbiotic stages of Gigaspora and Glomus endophytes were found. The flavonoid NMHTV isolated from non AM clover shoot, did not affect the percentage of germination of spores but significantly increased (P < 0.05) the other steps of the presymbiotic stage of Gi. margarita spores when 2 μM concentration was used. The symbiotic stage of Gi. margarita was also significantly increased when 2 μM of the flavonoid NMHTV was applied. This flavonoid had no effect on the presymbiotic development of G. mosseae, G. intraradices and Gi. rosea except when 8 μM concentration was used, which inhibited the hyphal length of Gi. rosea. These results suggest the possible implication of the flavonoid NMHTV in the susceptibility of tomato roots to the AM formation by Gi. margarita. The absence of stimulation of the AM presymbiotic and symbiotic stages in tomato by exogenous application of the newly synthesized flavonoids MH-1, MH-2, and MH-3, in clover shoots after AM colonization, indicated that the autorregulation of the AM symbiosis can be, at least partially, due to the disappearance of flavonoids in AM colonized plants that stimulated the AM symbiosis. Fil: Scervino, Jose Martin. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentina Fil: Ponce, María Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono; Argentina Fil: Della Mónica, Ivana Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono; Argentina Fil: Vierheilig, Horst. Universitat Fur Bodenkultur Wien; Austria. Consejo Superior de Investigaciones Científicas. Estación Experimental del Zaidín; España Fil: Ocampo, Juan Antonio. Consejo Superior de Investigaciones Científicas. Estación Experimental del Zaidín; España Fil: Godeas, Alicia Margarita. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Biodiversidad y Biología Experimental; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The effect of the flavonoids chrysin, isorhamnetin, kaempferol, luteolin, morin and rutin on pre-symbiotic growth, such as spore germination, hyphal length, hyphal branching and the formation of auxiliary cells and secondary spores, of the arbuscular mycorrhizal fungi Gigaspora rosea, G. margarita, Glomus mosseae and G. intraradices was studied. According to the effect on each fungal growth parameter, the tested compounds could be classified to be genus and/or species specific or specific, for a certain developmental stage of pre-symbiotic growth. A clear arbuscular mycorrhizal genus specific, and even species specific, effect of some flavonoids was observed. However, this specificity of a flavonoid could not be generalized but differs depending on the pre-symbiotic stage of the AM fungus. Moreover, our show that for a better understanding of the role of compounds in the AM symbiosis, studies should not be conducted only with one AM fungus looking at one fungal growth parameter such as spore germination or hyphal length, but should be wider, including several growth parameters and several AM fungi.

The influence of tillage and monoculture on arbuscular mycorrhizae (AM) fungal species diversity in a field site of the Buenos Aires province (Argentina) was investigated through the morphological characterization of AM spores. Glomalean spores were isolated by wet sieving and decanting from three plots cropped either to wheat, barley or clover and from a grassland at the margins of the cultivated plots. Seventeen different Glomalean species were detected overall and seven of them were identified. Total species number as well as spore and species richness found in grassland and clover soil were higher than those found in soils planted either with barley or wheat. The most frequently occurring species in the site were Glomus mosseae, Scutellospora pellucida, Glomus sp. 7 and Gigaspora sp. 1. The first three were also the most dominant species and were found in the four types of analysed soils. In grassland soil and wheat, the dominant species was Glomus sp. 6. S. pellucida was dominant in barley, and in red clover the dominant species were G. mosseae and S. pellucida. Tillage and cereal monoculture negatively affected diversity of AM fungal species. Natural re-colonization of indigenous AM fungi was observed in cultivated soil with red clover for 3 years, suggesting that this host could be used as a cover crop to increase AM fungal inocula in disturbed soils. Arbuscular mycorrhizal populations were associated with natural and cultivated vegetation on a site of Buenos Aires province, Argentina. Fil: Menendez, Ana Bernardina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; Argentina Fil: Scervino, Jose Martin. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Godeas, Alicia Margarita. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina