Formação de ectomicorrizas por monocários e dicários de Pisolithus sp. e interações nutricionais em Eucalyptus grandis Formation of ectomycorrhizae by monokaryons and dikaryons of Pisolithus sp. and nutritional interactions in Eucalyptus grandis

A germinação de basidiósporos de Pisolithus spp. dá origem a monocários, caracterizados por possuírem um único núcleo haplóide por célula. No campo, o eucalipto associa-se a micélios dicarióticos de Pisolithus spp., não havendo relatos sobre a capacidade dos monocários em estabelecer a associação ectomicorrízica com a planta hospedeira nessas condições. Embora os monocários de Pisolithus sp. sejam capazes de formar a associação ectomicorrízica in vitro, nada se sabe sobre a capacidade dessas estirpes em promover a absorção de nutrientes e o crescimento do eucalipto. O objetivo deste trabalho foi estudar a formação de ectomicorrizas por isolados monocarióticos e dicarióticos de Pisolithus sp. em Eucalyptus grandis, sob condições de casa de vegetação, bem como investigar as relações entre o estabelecimento da associação e o crescimento e a absorção de P, Ca, Mg, K, Cu e Zn pelas plantas. Caracterizou-se, também, a produção de massa seca micelial e a absorção de nutrientes pelos isolados fúngicos in vitro. Os isolados monocarióticos e dicarióticos testados variaram na produção de massa seca micelial e na capacidade de absorção de nutrientes. Em geral, os monocários apresentaram maiores índices de eficiência de utilização de nutrientes do que os dicários. Todos os isolados monocarióticos e dicarióticos formaram ectomicorrizas típicas quando associados com E. grandis. A presença dos isolados fúngicos monocarióticos associados às raízes laterais resultou em aumento do diâmetro radial das células da epiderme radicular, característico das ectomicorrizas de eucalipto, indicando que os monocários, à semelhança dos dicários, são capazes de produzir reguladores de crescimento. As médias de percentagem de colonização das raízes pelos isolados monocarióticos e dicarióticos variaram de 12 a 76 %. A absorção de Ca, Mg e K foi estimulada de forma expressiva pela presença das ectomicorrizas, com aumentos de até 760 vezes, demonstrando a relevância dessa associação no suprimento desses macronutrientes, especialmente o Ca. Alguns isolados monocarióticos são tão eficientes quanto os dicários na colonização radicular e na absorção de nutrientes. A caracterização dos monocários de Pisolithus sp. é necessária para se estabelecer a seleção e o cruzamento dos isolados com características desejáveis, visando ao melhoramento genético e à maior eficiência da associação simbiótica. Este trabalho constitui o primeiro relato das interações nutricionais entre monocários de Pisolithus sp. e a planta hospedeira.<br>The germination of Pisolithus spp. basidiospores originates monokaryons, characterized by having a single haploid nucleus per cell. In the field, eucalypts are associated with dykaryons of Pisolithus spp., there being no reports on the capacity of monokaryons to establish the ectomycorrhizal association with the host plant under such conditions. Although Pisolithus spp. monokaryons have been shown to form ectomycorrhizas in vitro, there is no information on the ability of these strains to promote nutrient uptake and growth of eucalypts. The objective of this study was to investigate the formation of ectomycorrhizae by monokaryotic and dikaryotic isolates of Pisolithus sp. in Eucalyptus grandis under greenhouse conditions, and the relationships between the establishment of ectomycorrhizae and the host plant growth and uptake of P, Ca, Mg, K, Cu, and Zn. The fungal isolates were highly variable in dry mycelial mass production and in their ability to take up macro- and micronutrients. Generally, the nutrient use efficiency of monokaryons was higher than that of dikaryons. All tested monokaryotic and dikaryotic isolates were able to form typical ectomycorrhizae when associated with E. grandis. The presence of monokaryotic isolates associated with the lateral roots of E. grandis resulted in typical increases in the radial diameter of the root epidermal cells, characteristic of eucalypt ectomycorrhizae, indicating that monokaryons, similarly to dikaryons, are capable of producing growth regulator compounds. The mean root colonization percentage of the monokaryotic and dikaryotic isolates varied from 12 to 76 %. The Ca, K, and Mg uptake was highly stimulated by the presence of ectomycorrhizae, with up to 760-fold increases, suggesting that the association must play a significant role in supplying the host with these nutrients, especially Ca. Some monokaryotic isolates are as efficient as the dikaryons at promoting root colonization and nutrient uptake. The characterization of Pisolithus sp. monokaryons is a basic requirement for the selection and crossing of isolates with desirable traits, aiming at the genetic improvement of fungal strains and a higher efficiency of the ectomycorrhizal association. This is the first report on the nutritional interactions between Pisolithus sp. monokaryons and the host plant

Subcellular Nutrient Element Localization and Enrichment in Ecto- and Arbuscular Mycorrhizas of Field-Grown Beech and Ash Trees Indicate Functional Differences

Mycorrhizas are the chief organ for plant mineral nutrient acquisition. In temperate, mixed forests, ash roots (Fraxinus excelsior) are colonized by arbuscular mycorrhizal fungi (AM) and beech roots (Fagus sylvatica) by ectomycorrhizal fungi (EcM). Knowledge on the functions of different mycorrhizal species that coexist in the same environment is scarce. The concentrations of nutrient elements in plant and fungal cells can inform on nutrient accessibility and interspecific differences of mycorrhizal life forms. Here, we hypothesized that mycorrhizal fungal species exhibit interspecific differences in mineral nutrient concentrations and that the differences correlate with the mineral nutrient concentrations of their associated root cells. Abundant mycorrhizal fungal species of mature beech and ash trees in a long-term undisturbed forest ecosystem were the EcM Lactarius subdulcis, Clavulina cristata and Cenococcum geophilum and the AM Glomus sp. Mineral nutrient subcellular localization and quantities of the mycorrhizas were analysed after non-aqueous sample preparation by electron dispersive X-ray transmission electron microscopy. Cenococcum geophilum contained the highest sulphur, Clavulina cristata the highest calcium levels, and Glomus, in which cations and P were generally high, exhibited the highest potassium levels. Lactarius subdulcis-associated root cells contained the highest phosphorus levels. The root cell concentrations of K, Mg and P were unrelated to those of the associated fungal structures, whereas S and Ca showed significant correlations between fungal and plant concentrations of those elements. Our results support profound interspecific differences for mineral nutrient acquisition among mycorrhizas formed by different fungal taxa. The lack of correlation between some plant and fungal nutrient element concentrations may reflect different retention of mineral nutrients in the fungal part of the symbiosis. High mineral concentrations, especially of potassium, in Glomus sp. suggest that the well-known influence of tree species on chemical soil properties may be related to their mycorrhizal associates