DNA barcoding as a tool for identification of plasmodia and sclerotia of myxomycetes (Myxogastria) appearing in moist chamber cultures
Shchepin ON, Dagamac NH, Sanchez OM, Novozhilov YK, Schnittler M, Zemlyanskaya IV 2017 -DNA barcoding as a tool for identification of plasmodia and sclerotia of myxomycetes (Myxogastria) appearing in moist chamber cultures. Abstract Moist chamber culture experiments are one of the basic methods of detection of myxomycete diversity that is usually employed to complement field datasets based on fruit bodies (sporocarps). However, often a large fraction of plasmodia that appear in moist chamber
... in moist chamber cultures does not yield sporocarps that can be determined to species based on morphological traits. Instead, plasmodia convert to a dormant stage called sclerotium. Both structures essentially lack taxonomically valuable morphological characters, preventing assignment to a species. Here we report the results of application of DNA barcoding as a method of taxonomical identification of plasmodia and sclerotia that develop in moist chamber cultures. The first ca. 600 bp of 18S rRNA gene were successfully amplified for 38 sclerotium and 32 plasmodium samples. Comparison to a large collection of reference sequences and phylogenetic analysis allowed identifying sequences up to species (45), genus (15) or order (10) following several formal criteria. Additionally four partial EF1A gene sequences were obtained, demonstrating that single-copy nuclear genes can also be easily amplified from plasmodia and sclerotia. The outlined methodology could facilitate future studies of myxomycete diversity and ecology based on moist chamber cultures, effectively increasing diversity estimates.