SYSTEMATICS OF MORCHELLA-ESCULENTA COMPLEX USING ENZYME-LINKED-IMMUNOSORBENT-ASSAY

Enzyme-linked Immunosorbent assay (ELISA) was used to study the immuno logical relationships among strains in the Morchella esculenta complex . Two strains of M. semilibera were chosen as outgroups for comparison . Mycelial extracts were used as antigens to induce antisera productio n in rabbits. The range of titers from ELISA were 1: 128 to 1:2048. Se rologic data from ELISA were analyzed based on the Titer and the Best- fit Line methods. Cross-reactivity at a specific titer did not differe ntiate the species. Dendrograms, however, based on the Best-fit Line m ethod had all strains of the M. esculenta complex cluster together and sort out separately from the M. semilibera strains. The Prager and Wi lson F values for the Titer method trees (UPGMA = 10.74, Fitch-Margoli ash = 17.89) were higher than those for the Best-fit Line method trees (UPGMA = 5.93, Fitch-Margoliash = 11.81) within each analysis indicat ing the trees from the Best-fit Line method were more parsimonious. No significant differences were found between UPGMA and the Fitch and Ma rgoliash method in terms of tree topology. Phylogenetic trees have not been commonly used with ELISA data. Combining the highly specific and sensitive ELISA with numerical taxonomic and phylogenetic techniques can provide additional systematic insights. The results of this study indicate that the gray, tan and large tan forms in the M. esculenta co mplex are immunologically indistinguishable and likely conspecific.