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Biologische Bundesanstalt für Land- und Forstwirtschaft, Institut für Pflanzenschutz im Obstbau, D-69221 Dossenheim, Germany
Correspondence
Erich Seemüller
E.Seemueller{at}bba.de
Apple proliferation (AP), pear decline (PD) and European stone fruit yellows (ESFY) are among the most economically important plant diseases that are caused by phytoplasmas. Phylogenetic analyses revealed that the 16S rDNA sequences of strains of each of these pathogens were identical or nearly identical. Differences between the three phytoplasmas ranged from 1·0 to 1·5 % of nucleotide positions and were thus below the recommended threshold of 2·5 % for assigning species rank to phytoplasmas under the provisional status Candidatus. However, supporting data for distinguishing the AP, PD and ESFY agents at the species level were obtained by examining other molecular markers, including the 16S23S rDNA spacer region, protein-encoding genes and randomly cloned DNA fragments. The three phytoplasmas also differed in serological comparisons and showed clear differences in vector transmission and host-range specificity. From these results, it can be concluded that the AP, PD and ESFY phytoplasmas are coherent but discrete taxa that can be distinguished at the putative species level, for which the names Candidatus Phytoplasma mali, Candidatus Phytoplasma pyri and Candidatus Phytoplasma prunorum, respectively, are proposed. Strains AP15R, PD1R and ESFY-G1R were selected as reference strains. Examination of available data on the peach yellow leaf roll (PYLR) phytoplasma, which clusters with the AP, PD and ESFY agents, confirmed previous results showing that it is related most closely to the PD pathogen. The two phytoplasmas share 99·6 % 16S rDNA sequence similarity. Significant differences were only observed in the sequence of a gene that encodes an immunodominant membrane protein. Until more information on this phytoplasma is available, it is proposed that the PYLR phytoplasma should be regarded as a subtype of Candidatus Phytoplasma pyri.
The GenBank/EMBL/DDBJ accession numbers for the P1/P7 amplimers (16S rRNA gene and 16S23S rDNA spacer region) of phytoplasma strains AP15R, AP1/93, PD1R, ESFY-G1R and ESFY-G2 are AJ542541, AJ542542, AJ542543, AJ542544 and AJ542545, respectively.
Restriction maps of rDNA obtained from P1/P7 PCR products from the apple proliferation, pear decline and European stone fruit yellows phytoplasmas are available as supplementary material in IJSEM Online.
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