Morphological and molecular analyses in Scleroderma species associated with some Caesalpinioid legumes, Dipterocarpaceae and Phyllanthaceae trees in southern Burkina Faso.

A combination of morphotypes, polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) analyses and internal transcribed spacer (ITS) sequencing was used to investigate Scleroderma species that were collected from woodlands in Burkina Faso. We harvested 52 specimens from 20 sites during rainy seasons between 1997 and 2000. According to their morphological features, these specimens were initially characterised, and we then identified six species of Scleroderma. Two of the species were clearly determined as Scleroderma dictyosporum Pat. and S. verrucosum Pers. The four remaining species were characteristically described as (1) displaying big spores with spines up to 2 microm (Scleroderma sp1), (2) producing spores without ornamentation (Scleroderma sp2), (3) spores with very small spines (Scleroderma sp3) and (4) with yellow sporocarps and sub-spherical spores (Scleroderma sp4). The specimens were then analysed using PCR/RFLP of the intergenic regions of rDNA, ITS and IGS1 and ITS sequencing. The restriction fragments obtained with two endonucleases, HinfI and MboI on ITS and IGS1 regions, showed that some isolates of S. dictyosporum had the same patterns as isolates and basidiocarps of Scleroderma sp4 (IR265, IR408, SP4-2903). Isolates of Scleroderma sp3 (IR252) had common restriction fragments as isolates of S. verrucosum (IR500, IR600). Intraspecific differences were observed in the two previously determined species, as well as in Scleroderma sp2. The ITS sequencing and phylogenetic analyses showed that the ribotypes identified by PCR/RFLP within these species might be phylogenetic species. Combining these molecular results allowed regrouping the six morphological species in three sets of cryptic species: a first set with two species including S. dictyosporum Pat., a second set with four species, including both S. verrucosum Pers. and Scleroderma sp1 and a third set with two species, including Scleroderma sp2. These investigations and the combined morphological and molecular analyses used to sort out species paved the way for identifying larger populations of Scleroderma species in Burkina Faso and other tropical zones.