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Specific detection of mycoplasma‐like organisms in European fruit trees by PCR 1
Author(s) -
JARAUSCH W.,
SAILLARD C.,
DOSBA F.,
BOVÉ J.M.
Publication year - 1995
Publication title -
eppo bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.327
H-Index - 36
eISSN - 1365-2338
pISSN - 0250-8052
DOI - 10.1111/j.1365-2338.1995.tb01461.x
Subject(s) - primer (cosmetics) , prunus , biology , malus , dna , polymerase chain reaction , microbiology and biotechnology , genetics , botany , gene , chemistry , organic chemistry
DNA amplification by PCR was employed to detect MLOs affecting Malus (apple) and Prunus (peach, apricot, plum) in Europe. Three primer pairs were chosen in a previously established sequence of a chromosomal fragment of apple proliferation MLO. Template DNA was extracted from experimentally inoculated trees and from symptomatic, naturally infected trees. Primer pair AP 5/4 specifically detected the DNA of apple proliferation MLO in infected trees resulting in a 483‐bp PCR product. Using primer pair AP 3/4, an MLO‐specific PCR product of 162 bp was obtained with total DNA extracted from MLO‐diseased Malus and Prunus. The PCR signal originating from Prunus MLOs was consistently weak, indicating a lower degree of homology to the primer. Therefore, nested PCR using external primer pair AP 5/6 in combination with internal primer pair AP 3/4 was applied to increase the sensitivity of detection of Prunus MLOs. Differentiation from apple proliferation MLOs was achieved using primer pair AP 5/4. No amplification with either primer pair could be obtained with DNA extracted from healthy plants.