APPENDIX 2 in The Amidella clade in Europe (Basidiomycota: Amanitaceae): clarification of the contentious Amanita valens (E.-J.Gilbert) Bertault and the importance of taxon-specific PCR primers for identification

APPENDIX 2. — Example of a matched comparison of Amanita ponderosa Malençon & R.Heim (ApoIf2-ITS4) and A. pseudovalens comb. nov., stat. nov. (ITS3- ApsIr3) for the same set of samples representing these two species, along with others of A. curtipes E.-J.Gilbert and A. lepiotoides Barla, and an unknown sample (SM-DB 61). The whole contents of each 20 µL PCR reactions were loaded, to help detect faint signals. Negative controls (neg.) with water instead of DNA. Amanita ponderosa is represented by the valens type (G73) and three samples from Spring 2010 (B07, P08, B10). Amanita pseudovalens is represented by the pseudovalens type (G74), the two MPU samples, Ode05 and Ode07. The Amanita curtipes samples (M03, M04 and M05) may produce a weak co-migrating band with the ITS3-ApsIR3 primer pair, and the A. lepiotoides samples (G08 and Eusk) were negative in both cases. Positive control reactions were not attempted because herbaria materials may be contaminated with other fungi.