Interação fungo micorrízico arbuscular e adubação fosfática no crescimento e absorção de nutrientes pelo porta-enxerto limoeiro Cravo

Estudou-se o efeito da inoculação com fungo micorrízico arbuscular (FMA), Glomus etunicatum, Glomus clarum, ou Scutelospora heterogama, e da adição de fósforo solúvel (15, 30, 45, 60, 120 e 240 mg de P kg-1 de terra) sobre as variáveis altura, número de folhas, diâmetro do caule, massas vegetais aérea e radicular, teores de nutrientes nas folhas e colonização micorrízica no porta-enxerto limoeiro Cravo (Citrus limonia (L.) Osbeck). Os FMAs, Glomus etunicatum e Glomus clarum, e doses crescentes de fósforo exerceram efeitos significativos sobre essas variáveis. O efeito da inoculação sobre as variáveis de crescimento vegetal foi mais acentuado nos tratamentos com adição de 30 a 60 mg de P kg-1 de terra. Para o teor de P foliar, o efeito da inoculação foi mais acentuado nos tratamentos adicionados de 120 a 240 mg de P kg-1 de terra. A inoculação com estes FMAs aumentou a eficiência do limoeiro Cravo em absorver nutrientes, tanto que as variáveis estudadas em plantas inoculadas na ausência de adubação fosfática superaram às de plantas não inoculadas em substrato adicionado de 240 mg de P kg-1 de terra.

PHOSPHORUS DOSES DETERMINE THE PREVALENCE OF NATIVE ARBUSCULAR MYCORRHIZAL FUNGI IN Araucaria angustifolia

A greenhouse experiment was installed with bait cultures to extract the AMF species present in a rhizosphere soil sample of a native Araucaria angustifolia forest in Campos do Jordao, Brazil. The experimental design was completely randomized, with four increasing phosphorus doses (0, 20, 50, and 150 mg kg(-1), as triple superphosphate), with five replicates, the bait plant was araucaria, and all pots were inoculated with 100 g of rhizospheric soil collected in an araucaria forest. After twelve months the spores were extracted, counted and identified, and the percent root colonization was also determined. When taking all four P doses into account, eleven AMF species could be identified: Acaulospora bireticulata, Acaulospora morrowiae, Acaulospora sp., Entrophospora colombiana, Gigaspora margarita, Glomus diaphanum, Glomus etunicatum, Glomus macrocarpum, Scutellospora calospora, Scutellospora gilmorei, and Scutellospora pellucida. There was no effect of the P dose on the total amount of spores neither on the percent root colonization. However, the correspondence analysis showed that the different AMF species were selectively associated mostly to either one or another P dose.

Phosphorus and Nitrogen Regulate Arbuscular Mycorrhizal Symbiosis in Petunia hybrida

Phosphorus and nitrogen are essential nutrient elements that are needed by plants in large amounts. The arbuscular mycorrhizal symbiosis between plants and soil fungi improves phosphorus and nitrogen acquisition under limiting conditions. On the other hand, these nutrients influence root colonization by mycorrhizal fungi and symbiotic functioning. This represents a feedback mechanism that allows plants to control the fungal symbiont depending on nutrient requirements and supply. Elevated phosphorus supply has previously been shown to exert strong inhibition of arbuscular mycorrhizal development. Here, we address to what extent inhibition by phosphorus is influenced by other nutritional pathways in the interaction between Petunia hybrida and R. irregularis. We show that phosphorus and nitrogen are the major nutritional determinants of the interaction. Interestingly, the symbiosis-promoting effect of nitrogen starvation dominantly overruled the suppressive effect of high phosphorus nutrition onto arbuscular mycorrhiza, suggesting that plants promote the symbiosis as long as they are limited by one of the two major nutrients. Our results also show that in a given pair of symbiotic partners (Petunia hybrida and R. irregularis), the entire range from mutually symbiotic to parasitic can be observed depending on the nutritional conditions. Taken together, these results reveal complex nutritional feedback mechanisms in the control of root colonization by arbuscular mycorrhizal fungi.

Early responses of wild plant seedlings to arbuscular mycorrhizal fungi and pathogens

Abstract Many plants form associations with arbuscular mycorrhizal fungi (AMF) because they profit from improved phosphorus nutrition and from protection against pathogens. Whereas mycorrhiza-induced pathogen protection is well understood in agricultural plant species, it is rarely studied in wild plants. As many pathogens infest plants in the first days after germination, mycorrhiza-induced pathogen protection may be especially important in the first few weeks of plant establishment. Here, we investigated interacting effects of {AMF} and the seedling pathogen Pythium ultimum on the performance of six- to seven-week-old seedlings of six wild plant species of the family Asteraceae in a full factorial experiment. Plant species differed in their response to AMF, the pathogen and their interactions. {AMF} increased and the pathogen decreased plant biomass in one and three species, respectively. Two plant species were negatively affected by {AMF} in the absence, but positively or not affected in the presence of the pathogen, indicating protection by AMF. This mycorrhiza-induced pathogen protection is especially surprising as we could not detect mycorrhizal structure in the roots of any of the plants. Our results show that even seedlings without established intraradical hyphal network can profit from AMF, both in terms of growth promotion in the absence of a pathogen and pathogen protection. The function of {AMF} is highly species-specific, but tends to be similar for more closely related plant species, suggesting a phylogenetic component of mycorrhizal function. Further studies should test a wider range of plant species, as our study was restricted to one plant family, and investigate whether plants profit from early mycorrhizal benefits in the long term.

Initial vegetation, and spore density, diversity and evenness of different arbuscular mycorrhizal fungi morphotaxa 29,36 and 39 months after experimental vegetation removal, central Argentina

1. Dominant plant functional types (PFTs) are expected to be primary determinants of communities of other above- and below-ground organisms. Here, we report the effects of the experimental removal of different PFTs on arbuscular mycorrhizal fungi (AMF) communities in a shrubland ecosystem in central Argentina. 2. On the basis of the biomass-ratio hypothesis and plant resource use strategy theory, we expected the effect of removal of PFTs on AMF colonization and spores to be proportional to the biomass removed and to be stronger when more conservative PFTs were removed. The treatments applied were: undisturbed control (no plant removed), disturbed control (mechanical disturbance), no shrub (removal of deciduous shrubs), no perennial forb (removal of perennial forbs), no graminoid (removal of graminoids) and no annual forb (removal of annual forbs). AMF colonization was assessed after 5,17 and 29 months. Total density of AMF spores, richness and evenness of morpho-taxa, and AMF functional groups were quantified after 5,17,29,36 and 39 months. 3. Five months after the initial removal we found a significant reduction in total AMF colonization in all plots subjected to PFT removals and in the disturbed control plots, as compared with the undisturbed controls. This effect disappeared afterwards and no subsequent effect on total colonization and colonization by arbuscules was observed. In contrast, a significant increase in colonization by vesicles was observed in months 17 and 29, mainly in no graminoid plots. In general, treatments did not significantly affect AMF spores in the soil. On the other hand, no annual forb promoted transient (12-18 months) higher ammonia availability, and no shrub promoted lower nitrate availability in the longer term (24-28 months). 4. Synthesis. Our experiment, the first to investigate the effects of the removal of different PFTs on AMF communities in natural ecosystems, indicates that AMF communities are resilient to changes in the soil and in the functional composition of vegetation. Furthermore, it does not provide consistent evidence in support of the biomass-ratio hypothesis or differential trait-based direct or indirect effects of different PFTs on AMF in this particular system.

Intracellular pH in Arbuscular Mycorrhizal Fungi1 : A Symbiotic Physiological Marker

A method was developed to perform real-time analysis of cytosolic pH of arbuscular mycorrhizal fungi in culture using dye and ratiometric measurements (490/450 nm excitations). The study was mainly performed using photometric analysis, although some data were confirmed using image analysis. The use of nigericin allowed an in vivo calibration. Experimental parameters such as loading time and concentration of the dye were determined so that pH measurements could be made for a steady-state period on viable cells. A characteristic pH profile was observed along hyphae. For Gigaspora margarita, the pH of the tip (0–2 μm) was typically 6.7, increased sharply to 7.0 behind this region (9.5 μm), and decreased over the next 250 μm to a constant value of 6.6. A similar pattern was obtained for Glomus intraradices. The pH profile of G. margarita germ tubes was higher when cultured in the presence of carrot (Daucus carota) hairy roots (nonmycorrhizal). Similarly, extraradical hyphae of G. intraradices had a higher apical pH than the germ tubes. The use of a paper layer to prevent the mycorrhizal roots from being in direct contact with the medium selected hyphae with an even higher cytosolic pH. Results suggest that this method could be useful as a bioassay for studying signal perception and/or H+ cotransport of nutrients by arbuscular mycorrhizal hyphae.

Congruence of fatty acid methyl ester profiles and morphological characters of arbuscular mycorrhizal fungi in Gigasporaceae.

Arbuscular mycorrhizal (AM) fungi (Order Glomales, Class Zygomycetes) are a diverse group of soil fungi that form mutualistic associations with the roots of most species of higher plants. Despite intensive study over the past 25 years, the phylogenetic relationships among AM fungi, and thus many details of evolution of the symbiosis, remain unclear. Cladistic analysis was performed on fatty acid methyl ester (FAME) profiles of 15 species in Gigaspora and Scutellospora (family Gigasporaceae) by using a restricted maximum likelihood approach of continuous character data. Results were compared to a parsimony analysis of spore morphological characters of the same species. Only one tree was generated from each character set. Morphological and developmental data suggest that species with the simplest spore types are ancestral whereas those with complicated inner wall structures are derived. Spores of those species having a complex wall structure pass through stages of development identical to the mature stages of simpler spores, suggesting a pattern of classical Haeckelian recapitulation in evolution of spore characters. Analysis of FAME profiles supported this hypothesis when Glomus leptotichum was used as the outgroup. However, when Glomus etunicatum was chosen as the outgroup, the polarity of the entire tree was reversed. Our results suggest that FAME profiles contain useful information and provide independent criteria for generating phylogenetic hypotheses in AM fungi. The maximum likelihood approach to analyzing FAME profiles also may prove useful for many other groups of organisms in which profiles are empirically shown to be stable and heritable.

Reciprocal N ((NH4+)-N-15 or (NO3-)-N-15) transfer between nonN(2)-fixing Eucalyptus maculata and N-2-fixing Casuarina cunninghamiana linked by the ectomycorrhizal fungus Pisolithus sp.

Two-way N transfers mediated by Pisolithus sp. were examined by excluding root contact and supplying (NH4+)-N-15 or (NO3-)-N-15 to 6-month-old Eucalyptus maculata or Casuarina cunninghamiana grown in two-chambered-pots separated by 37 m screens. Mycorrhizal colonization was 35% in Eucalyptus and 66% in Casuarina (c. 29% N-2-fixation). Using an environmental scanning electron microscope, living hyphae were observed to interconnect Eucalyptus and Casuarina. Biomass and N accumulation was greatest in nodulated mycorrhizal Casuarina/mycorrhizal Eucalyptus pairs, less in nonnodulated mycorrhizal Casuarina/mycorrhizal Eucalyptus pairs, and least in nonnodulated nonmycorrhizal Casuarina/nonmycorrhizal Eucalyptus pairs. In nonnodulated mycorrhizal pairs, N transfers to Eucalyptus or to Casuarina were similar (2.4-4.1 mg per plant in either direction) and were 2.6-4.0 times greater than in nonnodulated nonmycorrhizal pairs. In nodulated mycorrhizal pairs, N transfers were greater to Eucalyptus (5-7 times) and to Casuarina (12-18 times) than in nonnodulated mycorrhizal pairs. Net transfer to Eucalyptus or to Casuarina was low in both nonnodulated nonmycorrhizal (< 0.7 mg per plant) and nonnodulated mycorrhizal pairs (< 1.1 mg per plant). In nodulated mycorrhizal pairs, net transfer to Casuarina was 26.0 mg per plant. The amount and direction of two-way mycorrhiza-mediated N transfer was increased by the presence of Pisolithus sp. and Frankia, resulting in a net N transfer from low-N-demanding Eucalyptus to high-N-demanding Casuarina.