Rapid detection and identification of ‘Candidatus Phytoplasma pini’-related strains (16SrXXI group)

What is it about?

In this communication, we report a ‘Ca. Phytoplasma pini’ 6,298 bp genomic DNA fragment that contains a partial DNA-dependent RNA polymerase Rpo’ gene (rpoC), a ribosomal protein Rps12 gene (rpsL), a ribosomal protein Rps7 gene (rpsG), an elongation factor EF-G gene (fusA), a translation elongation factor EF-TU gene (tuf), and a gene encoding a hypothetical protein (a DAK2 domain fusion protein YloV). For routine application in disease surveys by foresters and quarantine organizations, we propose a new approach to detect ‘Ca. Phytoplasma pini’ based on 16S rRNA gene and tuf gene amplification in direct PCRs.

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Photo by IvanH on Unsplash

Photo by IvanH on Unsplash

Why is it important?

Advantageously, the PCR protocols that we developed in the current study offer both increased sensitivity and selectivity as we used ‘Ca. Phytoplasma pini’-unique sequences in primer design for both 16S rDNA- and tuf gene-based assays. Our new ‘Ca. Phytoplasma pini’ detection protocols effectively eliminate the need to perform nested-PCR nor post-PCR RFLP analysis. Since the new assays significantly reduce the time and operational cost of ‘Ca. Phytoplasma pini’ detection, they are especially desirable in large-scale surveillance studies, helping researchers, foresters and pine tree growers detect and identify ‘Ca. Phytoplasma pini’ and related strains more easily and rapidly.

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