Methylome Analysis of Two Xanthomonas spp. Using Single-Molecule Real-Time Sequencing

• Published in: The plant pathology journal Vol. 32; no. 6; pp. 500 - 507
• Main Authors: Seong, Hoon Je, Park, Hye-Jee, Hong, Eunji, Lee, Sung Chul, Sul, Woo Jun, Han, Sang-Wook
• Format: Journal Article
• Language: English
• Published: Korea (South) 한국식물병리학회 01.12.2016; Korean Society of Plant Pathology; Hanrimwon Publishing Company
• Subjects: methylome; single-molecule real-time sequencing; Xanthomonas; 농학; methylome; single-molecule real-time sequencing; Xanthomonas
• ISSN: 1598-2254; 2093-9280
• DOI: 10.5423/ppj.ft.10.2016.0216

Single-molecule real-time (SMRT) sequencing allows identification of methylated DNA bases and methylation patterns/motifs at the genome level. Using SMRT sequencing, diverse bacterial methylomes including those of , spp., and have been determined, and previously unreported DNA methylation motifs have been identified. However, the methylomes of species, which belong to the most important plant pathogenic bacterial genus, have not been documented. Here, we report the methylomes of pv. (Xag) strain 8ra and pv. (Xcv) strain 85-10. We identified N -methyladenine (6mA) and N -methylcytosine (4mC) modification in both genomes. In addition, we assigned putative DNA methylation motifs including previously unreported methylation motifs via REBASE and MotifMaker, and compared methylation patterns in both species. Although Xag and Xcv belong to the same genus, their methylation patterns were dramatically different. The number of 4mC DNA bases in Xag (66,682) was significantly higher (29 fold) than in Xcv (2,321). In contrast, the number of 6mA DNA bases (4,147) in Xag was comparable to the number in Xcv (5,491). Strikingly, there were no common or shared motifs in the 10 most frequently methylated motifs of both strains, indicating they possess unique species- or strain-specific methylation motifs. Among the 20 most frequent motifs from both strains, for 9 motifs at least 1% of the methylated bases were located in putative promoter regions. Methylome analysis by SMRT sequencing technology is the first step toward understanding the biology and functions of DNA methylation in this genus.