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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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.
Perspectives
The supplemental use of such a single copy gene, which is more variable in nucleotide sequence than the 16S rRNA gene, provides a means for differentiating genotypes present among ‘Ca. Phytoplasma pini’ strains. Together the use of both 16S rRNA gene and tuf gene analysis should aid searches for the as-yet-unknown insect-vector(s) responsible for spread of ‘Ca. Phytoplasma pini’ strains that are damaging pine forests in Europe.
Dr. Deividas Valiunas
Nature Research Centre, Akademijos St. 2, LT-08412 Vilnius, Lithuania
Read the Original
This page is a summary of: Rapid detection and identification of ‘Candidatus
Phytoplasma pini’-related strains based on genomic markers present in 16S rRNA and tuf
genes, Forest Pathology, September 2019, Wiley,
DOI: 10.1111/efp.12553.
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