Molecular characterization of Xanthomonas species isolated from Araceae and the development of a triplex TaqMan assay for detection of Xanthomonas phaseoli pv. dieffenbachiae

J.M. van der Wolf*, M.C. Krijger, O. Mendes, B. Brankovics, P.J.M. Bonants, Leonie Didden, Ellis Meekes

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

1 Citation (Scopus)

Abstract

In total 58 Xanthomonas strains isolated from Araceae worldwide, together with 13 other phylogenetically-related Xanthomonas strains, were characterized using multilocus sequence analysis based on concatenated sequences of seven single copy orthologous genes, extracted from whole genome sequences. The analysis revealed a monophyletic clade of 48 strains, 44 isolated from Anthurium, identified as X. phaseoli pv. dieffenbachiae (Xpd) confirmed by nucleotide identity analysis. The other strains from aroids were identified as Xanthomonas euvesicatoria (2 strains), X citri (5 strains) and Xanthomonas sacchari (3 strains). Two TaqMan assays were designed for specific detection of Xpd, one targeting sequences of a hypothetical protein and one targeting a type I restriction endonuclease subunit S. The two assays showed similar reaction kinetics and were merged with an assay comprising an amplification and extraction control into a triplex assay. The assay was able to detect minimally 100 copies of a target sequence delivered as a gBlock, 100 fg of genomic DNA and 104 cells per mL in an Anthurium leaf extract.
Original languageEnglish
Pages (from-to)167-179
Number of pages13
JournalEuropean Journal of Plant Pathology
Volume163
Issue number1
Early online date22 Mar 2022
DOIs
Publication statusPublished - May 2022

Keywords

  • Xanthomonas axonopodis pv. dieffenbachiae
  • Multilocus sequence analysis
  • Whole genome sequencing
  • Anthurium
  • Average nucleotide identity
  • Taxonomy

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