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Phytopathology. 2018 Dec 27. doi: 10.1094/PHYTO-09-18-0326-R. [Epub ahead of print]

Multilocus characterization, gene expression analysis of putative immunodominant protein coding regions, and development of recombinase polymerase amplification assay for detection of 'Candidatus Phytoplasma pruni' in Prunus avium.

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Washington State University, Plant Pathology, Prosser, Washington, United States ;
Washington State University, Plant Pathology, Prosser, Washington, United States ;


Western X (WX) disease, caused by 'Candidatus Phytoplasma pruni', is a devastating disease of sweet cherry resulting in the production of small, bitter flavored fruits that are unmarketable. Recent escalation of WX disease in Washington State prompted the development of a rapid detection assay based on recombinase-polymerase amplification (RPA) to facilitate timely removal and replacement of diseased trees. Here, we report a reliable RPA assay targeting putative immunodominant protein coding regions that showed comparable sensitivity to PCR in detecting 'Ca. Phytoplasma pruni' from crude sap of sweet cherry tissues. Apart from the predominant strain of 'Ca. Phytoplasma pruni', the RPA assay also detected a novel strain of phytoplasma from several WX affected trees. Multilocus sequence analyses using the idpA, imp, rpoE, secY and 16S rRNA regions from several 'Ca. Phytoplasma pruni' isolates from WX affected trees showed that this novel phytoplasma strain represents a new subgroup within the 16SrIII group. Examination of high throughput sequencing data from total RNA of WX affected trees revealed that the imp coding region is highly expressed, and, as supported by quantitative RT-PCR data, showed higher RNA transcript levels than the previously proposed immunodominant protein A (idpA) coding region of 'Ca. Phytoplasma pruni'.


Bacteriology; Disease control and pest management; Population biology


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