Abstract
The species Acidovorax avenae contains bacteria pathogenic to several crops: A. a. subsp. avenae infects oats, corn, rice, millet, and sugarcane; subsp. citrulli infects watermelon and melon; and subsp. cattleyae infects orchid. Because these bacteria are closely related their proper identification requires expensive, time-consuming pathogenicity tests. To develop PCR assays for rapid identification of all species and subsp. citrulli, we sequenced the intergenic spacer region of the 16S–23S rDNA and designed avenae-specific and subsp. citrulli-specific classical primers and real-time primers and probes. Classical avenae-specific primers Oafl/Oarl amplified a DNA product from all 46 strains of subsp. avenae originating from foxtail, oats, corn, rice, sugarcane, and millet; the type strains of subsp. cattleyae from orchid; and 11 strains of subsp. citrulli from watermelon. Real-time avenae-specific primers Oafl/Oar2 and probe AaP1 reacted with all 40 strains of subsp. avenae, the type strain of subsp. cattleyae, and 8 strains of subsp. citrulli. Classical citrulli-specific primers Aacf2/Aacr2 and real-time primers Aacf3/Aacr2 and probe AaP2 amplified a PCR product from all 11 strains of subsp. citrulli, but not from any other strain of subsp. avenae. None of over 50 other bacteria tested generated a PCR product with either classical primers or real-time primers and probe. The species-specific classical primers Oafl/Oarl and real-time primers Oafl/Oar2 and probe AaP1 should prove useful for rapid identification of A. avenae strains from all hosts, whereas the citrulli-specific classical primers Aacf2/Aacr2 and real-time primers Aacf3/Aacr2 and probe AaP2 should be useful for specific identification of subsp. citrulli.
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Song, W.Y., Sechler, A.J., Hatziloukas, E., Kim, H.M., Schaad, N.W. (2003). Use of PCR for Rapid Identification of Acidovorax avenae and A. avenae subsp. citrulli . In: Iacobellis, N.S., et al. Pseudomonas syringae and related pathogens. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-0133-4_59
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DOI: https://doi.org/10.1007/978-94-017-0133-4_59
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