KMID : 0545120190290050785
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Journal of Microbiology and Biotechnology 2019 Volume.29 No. 5 p.785 ~ p.793
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Development of Molecular Marker through Genome Realignment for Specific Detection of Xanthomonas campestris pv. campestris Race 5, a Pathogen of Black Rot Disease
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Afrin Khandker Shazia
Rahim Md Abdur Jung Hee-Jeong Park Jong-In Kim Hoy-Taek Nou Ill-Sup
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Abstract
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Black rot caused by Xanthomonas campestris pv. campestris (Xcc) is the most damaging disease in Brassica crops around the world. In this study, we developed a molecular marker specific to Xcc race 5. To do this, the available whole genome sequences of Xcc races/strains and Xc subspecies were aligned and identified a highly variable genomic region (XccR5-89.2). Subsequently, a primer set covering the ¡®XccR5-89.2¡¯ region was designed and tested against the genomic DNA of Xcc races/strains, Xc subspecies and other plant-infecting bacterial strains (Pseudomonas syringae pv. maculicola and Erwinia carotovora subsp. carotovora). The results showed that the ¡®XccR5-89.2¡¯ primer pair amplified a 2,172-bp fragment specific to Xcc race 5. Moreover, they also amplified a 1,515-bp fragment for Xcc race 1 and an over 3,000-bp fragment for Xcc race 3. However, they did not amplify any fragments from the remaining Xcc races/strains, subspecies or other bacterial strains. The ¡®XccR5-89.2¡¯ primer pair was further PCR amplified from race-unknown Xcc strains and ICMP8 was identified as race 5 among nine race-unknown Xcc strains. Further cloning and sequencing of the bands amplified from race 5 and ICMP8 with ¡®XccR5-89.2¡¯ primers revealed both carrying identical sequences. The results showed that the ¡®XccR5-89.2¡¯ marker can effectively and proficiently detect, and identify Xcc race 5 from Xcc races/strains, subspecies and other plant-infecting bacteria. To our knowledge, this is the first report for an Xcc race 5-specific molecular marker.
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KEYWORD
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Xanthomonas campestris pv. campestris, race 5, black rot, molecular marker, cabbage
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