The present study aims at providing standard values for the exploration type (ET)-specific quantification of extramatrical mycelium (EMM) of ectomycorrhizal fungi applicable to ecological field studies. These values were established from mycelial systems of ectomycorrhizae (ECM) synthesized in rhizotrons with near-to-natural peat substrate. Based on image analysis, the "Specific Potential Mycelial Space Occupation" (sPMSO), i.e. the ET-specific complete area that is covered by the EMM systems (mm 2 cm −1 ECM −1 ), and the "Specific Actual Mycelial Space Occupation" (sAMSO), i.e. the projection area of mycelial systems (mm 2 cm −1 ECM −1 ), were analyzed as an extension of a previously described approach. The "Specific Extramatrical Mycelial Length" (sEML) [m cm −1 ECM −1 ] and the "Specific Extramatrical Mycelial Biomass" (sEMB) (μg cm −1 ECM −1 ) were calculated for each of the ET via the proportion of hyphal projected area, hyphal length and biomass, the latter two being derived from previous measurements on Piloderma croceum, a "Medium-Distance" (MD)-ET. Both sPMSO and sAMSO were highest for the "Long-Distance" (LD)-ET, whereas those of the "Short-Distance" (SD)-ET and MD-ET were similar, although showing high variation. In contrast, mycelial density per occupied area of the MD-ET was twice as high as that of the LD-ET. Proportional to the sAMSO, the EMM length and biomass differed considerably between the three ET with values of the MD-ET being 1.9 times higher than those of SD-ET, and those of the LD-ET being 15 times higher than those of the SD-ET. These standards in relation to ECM length may ease quantification of mycelial space occupation and biomass in a relatively simple way. Thereby, the ET-specific contribution of EMM can be distinguishedalso of non-cultivable species-and up-scaling to large-scale estimation of cost/benefit relations is possible.
DNA-based quantification methods such as real-time TaqMan PCR allow a rapid and highly sensitive species-specific quantification of isolated fungal DNA material, but most quantification systems are only able to measure relative amounts of biomass or biomass changes during different treatments. In this experiment, an already established DNA quantification system for the ectomycorrhizal fungus Piloderma croceum, based on the ITS region of ribosomal DNA, was calibrated to absolute biomass to obtain a direct correlation between mycelial biomass and isolated ITS copies. Thin layers of sterile mycelia were cultured on slides. The mycelial biomass was calculated from measurements of the total hyphal length using image analysis, followed by determination of the mycelial volume, and multiplied by the specific weight of hyphae obtained from literature data. Using the very same mycelium, the number of ITS copies was quantified by TaqMan PCR. The mean value of 1047 (+/- 185) copies per mm hypha results in possible data for a direct conversion: one billion (10 (9)) ITS copies corresponded to 0.79 mg hyphal dry weight. For the ribosomal ITS multi-copy genes, the number of ITS copies could be calculated to approx. 152 (+/- 26) copies per dikaryotic cell. These conversion data now allow determination of the mycelial biomass of Piloderma croceum using real-time TaqMan PCR, a prerequisite for competition experiments with Piloderma croceum.
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