Obtaining positive controls for the detection of two polymorphic regions in Cuban strains of ‘Candidatus Liberibacter asiaticus’
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Abstract
“Huanglongbing” (HLB) is currently the most destructive disease of citrus worldwide. The detection, identification and genetic diversity studies of strains of its main associated agent ‘Candidatus Liberibacter asiaticus’ were limited to the use of housekeeping genes. Because of the increasing availability of complete genome sequences of different ‘Ca. L. asiaticus’ strains from different countries, regions inside the genome of these bacteria having greater variability have been successfully implemented for its characterization, including microsatellites, prophage genes and miniature inverted-repeat transposable elements (MITEs). In the present work, six Cuban strains of ‘Ca. L. asiaticus’ with different geographical origins (Western, Central and Eastern regions) were included. Two polymorphic markers (prophage types and MITEs) were used to verify the presence of genetic diversity among the strains. The combination of the information obtained from detecting markers allowed verifying the strain differentiation according to the length of the amplified bands. The detected strains are important controls necessary to guarantee the performance and interpretation of tests using these molecular markers. The PCR systems used will allow a fast and improved characterization of the bacterial populations present in Cuba. This is the first report of the detection of polymorphic regions in the genome of Cuban strains of ‘Ca. L. asiaticus’.
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