16S rDNA‐derived oligonucleotide probes for the differential diagnosis of plum leptonecrosis and apple proliferation phytoplasmas
- 28 June 1996
- journal article
- Published by Wiley in EPPO Bulletin
- Vol. 26 (2) , 421-428
- https://doi.org/10.1111/j.1365-2338.1996.tb00606.x
Abstract
The 16S rRNA gene of plum leptonecrosis phytoplasma has been PCR‐amplified, cloned and almost completely sequenced (1201 bp). The sequence analysis confirmed the close genetic relationship between plum leptonecrosis phytoplasma and the phytoplasmas associated with other stone‐fruit diseases in Europe. By comparison with the 16S rDNA sequence of apple proliferation phytoplasma, two oligonucleotides were selected, differing by two nucleotides, which were specific for apple proliferation and plum leptonecrosis phytoplasmas, respectively. The oligonucleotides were labelled with digoxigenin and hybridized, in the presence of tetramethylammonium chloride, to 16S rDNA fragments amplified from apple and plum leaf samples. The results showed that, under the described hybridization conditions, the two phytoplasmas could clearly be distinguished. The advantage of the proposed technique over 16S rDNA restriction fragment length polymorphism is discussed.Keywords
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