Diversity of the ectomycorrhiza community at a uranium mining heap

Ectomycorrhiza (EM) community structure was analyzed at one bare heap site (BHS), one heap site with organic cover (HS-OH) and one reference site (RS) in the former uranium mining area near Ronneburg (Thuringia, Germany). Twenty-three EM morphotypes were distinguished, and 14 of them were additionally characterized by polymerase chain reaction (PCR), restriction fragment length polymorphism (RFLP) and internal transcribed spacer (ITS) sequence analysis. Colonization of birch by the different morphotypes was quantified, and the EM diversity at the different sites was investigated. Compared to RS, total EM colonization was reduced by 6% (P=0.851) at HS-OH and by 58% (P<0.001) at BHS. Likewise, EM diversity was reduced by 16% (P=0.229) at HS-OH and 52% (P<0.001) at BHS. The Sørensen similarity between EM samples from RS was nearly independent from the sampling date, whereas at HS-OH and especially BHS, the Sørensen similarity decreased with increasing time between the samplings. All EM fungal species dominating at the two heap sites were also present at RS. Thus, fungi with high tolerance against uranium and other stress factors at the heap sites (e.g. heavy metals, nutrient limitation, drought) were selected among all EM fungi of the area. Highly adapted fungi with a distribution restricted to the contaminated soils were not detected.Electronic Supplementary Material Supplementary material is available for this article at .     

Ectomycorrhizal fungi identification in single and pooled root samples: terminal restriction fragment length polymorphism (TRFLP) and morphotyping compared

We describe here the results of a study conducted to evaluate a terminal restriction fragment length polymorphism (TRFLP) approach targeting rRNA genes for determination of ectomycorrhizal (ECM) communities colonizing the roots of loblolly pine (Pinus taeda L.). Root tips separated from soil cores were classified according to morphological characteristics and DNA was then extracted from each group of morphotyped tips. Labeled primers were used to generate terminal restriction fragments (TRF) for molecular fingerprinting of root colonizing fungi and to determine how well TRFLP could be used to discriminate between ectomycorrhizal types. Morphotypes generally correlated well with specific TRFs and sequence analysis confirmed that TRFs could be used to discriminate among fungal types. Sequence analysis indicated that important ECM fungi including Russulaceae, Thelephorales, and Tricholomataceae could be fingerprinted with TRFLP. In addition, a fixed proportion of the DNA extracted from each morphotype from the same core was used in a pooling experiment used to assess whether previously identified fungal species types could be distinguished from one another within reconstructed communities. Since some morphotypes share TRFs, dual analysis of ITS1 and ITS2 was necessary for accurate fingerprinting of fungal types. Approximately, 5.0±0.3 phylotypes were detected per core with TRFLP-corrected morphotyping as compared to 4.0±0.4 phylotypes using TRFLP on pooled community samples. TRFLP made on experimental sporocarp communities suggested that reduced ECM richness with TRFLP may be partly caused by differences in the amount of DNA available for PCR and primer bias. Nonetheless, TRFLP on pooled morphotypes accounted for more than 93% of colonized root tips. The method can be used to facilitate analysis of mycorrhizal communities using root tips collected from soil cores.