Publications

4787

Abstract The Genome Taxonomy Database (GTDB; https://gtdb.ecogenomic.org) provides a phylogenetically consistent and rank normalized genome-based taxonomy for prokaryotic genomes sourced from the NCBI Assembly database. GTDB release 10 (R10-RS226) spans 715 230 bacterial and 17 245 archaeal genomes organized into 136 646 bacterial and 6968 archaeal species clusters. Fewer new major branches of prokaryotic life are being discovered with each release of GTDB, suggesting that we are beginning to saturate readily discoverable microbial diversity through culture-independent analyses. However, species discovery continues unabated as >95% of bacterial and archaeal species remain to be genomically elucidated based on conservative projections. We present additions to the GTDB website, methodological improvements, policy changes, notable nomenclatural updates, and user applications. We conclude with a summary of future plans for the resource including a fungal taxonomy and a nomenclatural extension to classify pathogens.

ABSTRACT X‐disease, caused by strains in the 16SrIII‐A subgroup of ‘ Candidatus Phytoplasma pruni’, is a devastating disease of Prunus species (stone fruits). Multiple outbreaks of this disease have occurred across much of North America for more than a century, with the most recent one beginning around 2010 in the Pacific Northwest of the United States, causing severe damage to the stone fruit industry. Sensitive and specific detection of X‐disease is critical to prevent the propagation and spread of infected plant material; however, current PCR‐based detection workflows lack specificity to X‐disease and require one or more PCR assays followed by additional testing to confirm the species identity. In this study, two real‐time PCR assays, one targeting the secY gene and the other targeting the tuf gene, were developed for specific detection of X‐disease in stone fruits. The CPH‐TXD assay targeting the tuf gene provided high‐sensitivity target detection of as few as five target copies with significantly reduced cross‐reactivity compared to current methods. The CPH‐SXD assay targeting the secY gene detected a minimum of 30 target copies and showed no cross‐reaction with closely related strains. The repeatability and reproducibility of both assays were verified with acceptable results. Together, these assays provide efficient and high‐specificity detection methods to combat the growing threat of X‐disease in North America.