Ectomycorrhizal System Research Articles

Endofungal bacteria and ectomycorrhizal fungi synergistically promote the absorption of organic phosphorus in Pinus massoniana.

Ectomycorrhizal fungi (ECMFs) that are involved in phosphorus mobilisation and turnover have limited ability to mineralise phytate alone. The endofungal bacteria in the ectomycorrhizal fruiting body may contribute to achieving this ecological function of ECMFs. We investigated the synergistic effect and mechanisms of endofungal bacteria and ECMF Suillus grevillea on phytate mineralisation. The results showed that soluble phosphorus content in the combined system of endofungal bacterium Cedecea lapagei and S. grevillea was 1.8 times higher than the sum of C. lapagei and S. grevillea alone treatment under the phytate mineralisation experiment. The S. grevillea could first chemotactically assist C. lapagei in adhering to the surface of S. grevillea. Then, the mineralisation of phytate was synergistically promoted by increasing the biomass of C. lapagei and the phosphatase and phytase activities of S. grevillea. The expression of genes related to chemotaxis, colonisation, and proliferation of C. lapagei and genes related to phosphatase and phytase activity of S. grevillea was also significantly upregulated. Furthermore, in the pot experiment, we verified that there might exist a ternary symbiotic system in the natural forest in which endofungal bacteria and ECMFs could synergistically promote phytate uptake in the plant Pinus massoniana via the ectomycorrhizal system.

Endofungal Bacterial Microbiota Promotes the Absorption of Chelated Inorganic Phosphorus by Host Pine through the Ectomycorrhizal System.

Ectomycorrhizal fungi play an irreplaceable role in phosphorus cycling. However, ectomycorrhizal fungi have a limited ability to dissolve chelated inorganic phosphorus, which is the main component of soil phosphorus. Endofungal bacteria in ectomycorrhizal fruiting bodies are always closely related to the ecological function of ectomycorrhizal fungi. In this study, we explore endofungal bacteria in the fruiting body of Tylopilus neofelleus and their function during the absorption of chelated inorganic phosphorus by host pine through the ectomycorrhizal system. The results showed that the endofungal bacterial microbiota in the fruiting body of T. neofelleus might be related to the dissolution of chelated inorganic phosphorus in soil. The soluble phosphorus content in the combined system of T. neofelleus and endofungal bacteria Bacillus sp. strain B5 was five times higher than the sum of T. neofelleus-only treatment and Bacillus sp. strain B5-only treatment in the dissolution experiment of chelated inorganic phosphorus. The results showed that T. neofelleus not only promoted the proliferation of Bacillus sp. strain B5 in the combined system but also improved the expression of genes related to organic acid metabolism, as assesed by transcriptomic analysis. Lactic acid content was five times higher in the combined system than the sum of T. neofelleus-only treatment and Bacillus sp. strain B5-only treatment. Two essential genes related to lactate metabolism of Bacillus sp. strain B5, gapA and pckA, were significantly upregulated. Finally, in a pot experiment, we verified that T. neofelleus and Bacillus sp. strain B5 could synergistically promote the absorption of chelated inorganic phosphorus by Pinus sylvestris in a ternary symbiotic system. IMPORTANCE Ectomycorrhizal fungi (ECMF) have a limited ability to dissolve chelated inorganic phosphorus, which is the main component of soil phosphorus. In the natural environment, the extraradical hyphae of ECMF alone may not satisfy the phosphorus demand of the plant ectomycorrhizal system. In this study, our results innovatively show that the ectomycorrhizal system might be a ternary symbiont in which ectomycorrhizal fungi might recruit endofungal bacteria that could synergistically promote the mineralization of chelated inorganic phosphorus, which ultimately promotes plant phosphorus absorption by the ectomycorrhizal system.

Tilia platyphyllos Scop.– Tuber brumale Vittad. vs. T. platyphyllos Scop.– T. borchii Vittad. ectomycorrhizal systems: a comparison of structural and functional traits

Ectomycorrhizae are mutualistic associations of several species of fungi with higher plants. Their formation involves alterations in the morphology and cell structure of the plant root and fungal mycelium. These modifications are correlated with mRNA and protein synthesis in the two symbionts. To gain more information about structural and functional traits involved in ectomycorrhizal formation, two “in vitro” ectomycorrhizal systems, set up by the inoculation of Tilia platyphyllos Scop. roots with either Tuber brumale Vittad. or T. borchii Vittad. mycelia, were investigated. Different parameters such as, fungal volume ratio, fungal biomass, plant and fungal transcript levels, specific enzymes activity and protein patterns were evaluated. In T. platyphyllos– T. brumale ectomycorrhizal tissue all the molecular and morphometrical approaches revealed a higher fungal biomass, volume and transcript as well as higher fungal protein levels respect to the host plant, suggesting that the fungal genes and proteins are up regulated after the establishment of symbiosis. These results are completely divergent from that obtained in T. platyphyllos Scop.– T. borchii Vittad. ectomycorrhizal system, leading us to hypothesise a different role of the fungal partner in the mycorrhization process according to the species it belongs to.

Jasmonic acid affects mycorrhization of spruce seedlings with Laccaria laccata

Jasmonic acid (JA) was shown to be an important regulating molecule after wounding and pathogen attack, but it also conveys mycorrhizal signalling. It was therefore applied to the ectomycorrhizal system. Root growth patterns of 2-month-old spruce seedlings mycorrhized with Laccaria laccata after treatment with 0.5 μM JA, 5.0 μM JA, or none for the control, were studied using a paper-sandwich technique. Changes in plant weight, root length, the degree of branching, presence of root hairs, and infection parameters were monitored for 26 days. Data were analysed by means of Student’s t-test and ANOVA factorial. Shoot and root dry weights were significantly increased by mycorrhizal infection, especially when JA was applied. Lateral root formation and the presence of roots with hairs were dramatically decreased in the presence of the fungus. A synergistic effect of JA and mycorrhization on decrease of lateral root length was shown. The first mycorrhizal contact of fungal hyphae with roots was significantly accelerated after treatment with JA.

Determination of 15N-labelled ammonium and total nitrogen in plant and fungal systems using mass spectrometry

A selected ion-monitoring method to measure 15N-labelled ammonia in biological samples was improved to simplify sample handling, to obviate interference from ammonia due to the decomposition of glutamine and to allow the determination of total N. Ammonia is derivatized with pentafluorobenzoylchloride to yield pentafluorobenzamide which is analysed by gas chromatogrpahy/mass spectrometry after clean-up using disposable silicic acid columns. The sensitivity achieved when operating in the negative ion-chemical ionization mode was somewhat higher than when using electron-impact ionization. Use of methyl amine as an internal standard improved the accuracy and precision of the measurements. The method was applied to samples taken from an intact ectomycorrhizal system fed with 15N-labelled ammonium and used to determine patterns of N assimilation into ammonium, free amino acids and macromolecular compounds.

The mycorrhizal fungus Amanita muscaria induces chitinase activity in roots and in suspension-cultured cells of its host Picea abies

A cell-wall fraction of the mycorrhizal fungus Amanita muscaria increased the chitinase activity in suspension-cultured cells of spruce (Picea abies (L.) Karst.) which is a frequent host of Amanita muscaria in nature. Chitinase activity was also increased in roots of spruce trees upon incubation with the fungal elicitor. Non-induced levels of chitinase activity in spruce were higher in suspension cells than in roots whereas the elicitorinduced increase of chitinase activity was higher in roots. Treatment of cells with hormones (auxins and cytokinin) resulted in a severalfold depression of enzyme activity. However, the chitinase activity of hormone-treated as well as hormone-free cells showed an elicitor-induced increase. Suspension cells of spruce secreted a large amount of enzyme into the medium. It is postulated that chitinases released from the host cells in an ectomycorrhizal system partly degrade the fungal cell walls, thus possibly facilitating the exchange of metabolites between the symbionts.