AUTHOR=Pasookhush Phongthana , Usmani Asmatullah , Suwannahong Kowit , Palittapongarnpim Prasit , Rukseree Kamolchanok , Ariyachaokun Kanchiyaphat , Buates Sureemas , Siripattanapipong Suradej , Ajawatanawong Pravech TITLE=Single-Strand Conformation Polymorphism Fingerprint Method for Dictyostelids JOURNAL=Frontiers in Microbiology VOLUME=12 YEAR=2021 URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.708685 DOI=10.3389/fmicb.2021.708685 ISSN=1664-302X ABSTRACT=

Dictyostelid social amoebae are a highly diverse group of eukaryotic soil microbes that are valuable resources for biological research. Genetic diversity study of these organisms solely relies on molecular phylogenetics of the SSU rDNA gene, which is not ideal for large-scale genetic diversity study. Here, we designed a set of PCR–single-strand conformation polymorphism (SSCP) primers and optimized the SSCP fingerprint method for the screening of dictyostelids. The optimized SSCP condition required gel purification of the SSCP amplicons followed by electrophoresis using a 9% polyacrylamide gel under 4°C. We also tested the optimized SSCP procedure with 73 Thai isolates of dictyostelid that had the SSU rDNA gene sequences published. The SSCP fingerprint patterns were related to the genus-level taxonomy of dictyostelids, but the fingerprint dendrogram did not reflect the deep phylogeny. This method is rapid, cost-effective, and suitable for large-scale sample screening as compared with the phylogenetic analysis of the SSU rDNA gene sequences.