PCR Procedure for Detection and Identification of European Stone-fruit Yellows (ESFY) Phytoplasma on Trees of Prunus Species

1 Fruit Growing Research Institute, BG-4004 Plovdiv, Bulgaria
2 Agricultural Univesity, BG-4000 Plovdiv, Bulgaria


TOPCHIISKA, M. and D. SAKALIEVA, 2002. PCR procedure for detection and identification of european stone-fruit yellows (ESFY) phytoplasma on trees of Prunus species. Bulg. J. Agric. Sci., 8: 19–22

Phytoplasma DNA of phloem tissues was isolated from naturally-infected almond, apricot and peach trees with symptoms of: yellows, rosetting, leaf roll, premature defoliation, as well as disturbances in the growth of annual shoots, drying and dying.
By employing the methods of polymerase chain reaction (PCR) and restriction fragment length polymorphism (PFLP) and using the universal (fU5/rU3 and fP1/rP7) and group specific (fO1/rO1) primers, samples of 10 trees, containing DNA, were tested repeatedly. By RFLP analyses of the PCR amplified ribosomal DNA of the 16S rDNA gene sequences and by the restriction endonucleases RsaI, SspI and Ssp6I, it was proven that three almond trees (isolates: No. 5–5A, No. 6–10A, and No. 6–26A), four apricot trees (isolates: No. 1–11A, No. 1–18A, and No. 6–26A), and three peach trees (isolates: No. 6–15I, No. 19–19I, and No. 20–24I) were infected with phytoplasma ­ the causative agent of the European stone fruit yellows (ESFY) disease.
For tested plum, cherry and sour cherry trees, which did not contain phytoplasma DNA, the results obtained by PCR procedures were negative.
This is the first report on detection of ESFY phytoplasma by species of the Prunus genus in Bulgaria

Key words: phytoplasma, stone fruits, Prunus sp.
Abbreviations: PCR – polymerase chain reaction; RFLP – restrictian fragment length polymorphism