Growth of the Ectomycorrhizal Fungus Pisolithus Microcarpus in different nutritional conditions

The most important plant species employed in reforestation programs depend on ectomycorrhizal fungi for their establishment and growth. The exploitation of this symbiosis to improve forest productivity requires fungal inoculants in a large scale level. To develop such a technology it is necessary to define the optimal composition of the culture medium for each fungus. With these objectives in mind, the effect of the composition of the culture medium on biomass production of the ectomycorrhizal fungus Pisolithus microcarpus (isolate UFSC-Pt116) was studied. The original composition of two culture media, already employed for cultivation of ectomycorrhizal fungi, was submitted to several variations with the C/N ratio as the main variable. A variation of the Pridham-Gottlieb medium was the most efficient for the production of biomass. Therefore, it was submitted to a factorial assay where glucose, peptone and yeast extract components were the factors analyzed. Results showed that the glucose concentration may be increased up to 40 percent in order to promote higher biomass production. Peptone had a positive effect on this variable, whereas yeast extract promoted a deleterious effect. These results indicate that it is advisable to eliminate yeast extract from the medium and replace it with peptone prior to use.

Growth of the Ectomycorrhizal Fungus Pisolithus Microcarpus in different nutritional conditions.

The most important plant species employed in reforestation programs depend on ectomycorrhizal fungi for their establishment and growth. The exploitation of this symbiosis to improve forest productivity requires fungal inoculants in a large scale level. To develop such a technology it is necessary to define the optimal composition of the culture medium for each fungus. With these objectives in mind, the effect of the composition of the culture medium on biomass production of the ectomycorrhizal fungus Pisolithus microcarpus (isolate UFSC-Pt116) was studied. The original composition of two culture media, already employed for cultivation of ectomycorrhizal fungi, was submitted to several variations with the C/N ratio as the main variable. A variation of the Pridham-Gottlieb medium was the most efficient for the production of biomass. Therefore, it was submitted to a factorial assay where glucose, peptone and yeast extract components were the factors analyzed. Results showed that the glucose concentration may be increased up to 40 % in order to promote higher biomass production. Peptone had a positive effect on this variable, whereas yeast extract promoted a deleterious effect. These results indicate that it is advisable to eliminate yeast extract from the medium and replace it with peptone prior to use.

Utilization of rocks and ectomycorrhizal fungi to promote growth of eucalypt

The utilization of rocks as fertilizers is limited by their low solubility. However, solubilization may be achieved by some micro-organisms, such as ectomycorrhizal fungi (ECMf). The aim of this study was to evaluate the potential of seven isolates of ECMf to solubilize two rocks, alkaline breccia and granite, and to liberate potassium and phosphorus for Eucalyptus dunnii seedlings under greenhouse conditions. Fungal inoculants were produced in a peat-vermiculite-liquid medium mixture and added to the planting substrate at 10 percent. Rocks were ground up and added at 0.500 mg and 16.0 mg per plant, as a source of phosphorus and potassium, respectively. Other nutrients were added and E. dunnii seeds were sown. Control plants, non-inoculated, were fertilized with the same amount of phosphorus and potassium using soluble forms. After 90 days, the plant height, shoot dry weight, root length, phosphorus and potassium contents, and mycorrhizal colonization were evaluated. Alkaline breccia was more efficient than granite as a source of phosphorus and potassium for the plants, and may be an alternative to conventional fertilizers. Isolates UFSC-Pt22 (Pisolithus sp.) and UFSC-Pt186 (Pisolithus microcarpus) were the most efficient in promoting plant growth, mainly when combined with alkaline breccia to replace potassium and phosphorus fertilizers, respectively.

Species richness and seasonal abundance of ectomycorrhizal fungi in plantations of Eucalyptus dunnii and Pinus taeda in southern Brazil.

The abundance and diversity of ectomycorrhizal fungi (EMF) was assessed based on the collection of basidiocarps during 12 months comprising the spring of 1995, and the summer, autumn, and winter of 1996, in three stands of young, middle-aged, and rotation age plantations of Pinus taeda and Eucalyptus dunnii, in the state of Santa Catarina, southern Brazil. A total of 3,085 collections yielded 34 presumed EMF taxa in ten genera, including mushroom-like and sequestrate species. Fruiting patterns of EMF differed with host and season, and host specificity was apparent in some. The overall relative importance (RI) and the Shannon diversity index (H) suggested that stands of E. dunnii had a more diverse aboveground EMF community than those of P. taeda. Overall, species of Scleroderma and Laccaria were not only the most abundant but also had the highest biomass values. The results show that a small number of species of abundant biomass and a larger number of species of less-abundant biomass characterize each forest class.

A PCR/RFLP technique to characterize fungal species in Eucalyptus grandis Hill ex. Maiden ectomycorrhizas.

With the increasing awareness of the significance of mycorrhizas, research is focusing on studies to elucidate the contribution of the symbiosis to ecosystem dynamics. In this sense, molecular biology has acquired great significance. PCR/RFLP techniques were adapted to characterize ectomycorrhizal fungi associated with Eucalyptus grandis. The ITS region of the fungal rDNA from pure cultures and from of mycorrhizas synthesized in vitro was amplified. Primers NSA3/NLC2 were used followed by a nested reaction with primers ITS1F/NLB3. Amplicons were then digested with the enzymes MboI, HinfI and TaqI. Amplification resulted in a 1,000-bp fragment for basidiomycetes and a 1,500 bp fragment for Cenococcum geophillum (an ascomycete). There was no amplification of the plant DNA. The enzymes MboI and HinfI were more effective than TaqI, resulting in patterns of two to five fragments allowing the identification of the isolates both in culture and in mycorrhizas. HinfI allowed greater differentiation among the isolates and a higher number of polymorphisms. Restriction with TaqI resulted in too many fragments. Amplification efficiency for the fungal DNA was 64% in culture and 87% in mycorrhizas. The modified methodology represents a valuable tool to complement traditional approaches in ecosystem studies.