Olive leaf spot is a disease of olive (Olea europaea L.) caused by the fungal pathogen, Spilocea oleaginea Cast. Progeny derived from crosses among susceptible, resistant, and semiresistant parental lines were assessed in the field for 8 years and classified as either resistant or susceptible. DNA from some of the progeny of this segregating population was used to identify molecular markers linked to olive leaf spot disease using the randomly amplified polymorphic DNA (RAPD) technique and bulked segregant analysis (BSA). Two DNA bulks were constructed, each containing 13 progeny showing either resistance or susceptibility for the disease, and screened for polymorphisms using 100 primers. One primer produced two polymorphic bands, one of ≈700 base pairs (bp) from the susceptible bulk and the other of ≈780 bp from the resistant bulk. The 780 bp marker appeared in 70.6% of the segregating progeny and 100% of parents showing resistance to leaf spot disease, while the 700 bp marker appeared in 47.1% of the segregating progeny and 100% of the parents showing susceptibility. These markers can be used as screening tools in olive improvement programs.
A sequence-tagged site (STS) was developed to identify a genetic marker linked to resistance to olive leaf spot caused by the pathogen, Spilocea oleaginea (Cast) (syn. Cycloconium oleaginum Cast.). The STS was based on a randomly amplified polymorphic DNA (RAPD) marker of about 780 base pairs (bp) linked to olive leaf spot resistance. Several primer pairs were developed to flank the sequence, and one pair produced the expected polymorphism between resistant and susceptible individuals tested, and was used as an STS marker. This primer pair was tested against parents and 34 individuals from a population segregating for resistance to olive leaf spot, and 12 commercial olive (Olea europaea L.) cultivars showing various levels of resistance to the disease. The STS marker was present in 71.4% of the parents and progeny that were designated as resistant, and was absent in 87% of the parents and progeny showing susceptibility. These primers were also able to distinguish cultivars such as `Koroneiki' and `Leccino', that are reported to show resistance to olive leaf spot, from `Barouni' and `Mission', that are reported to be susceptible. This is the first report of a STS marker for olive, and its use will assist greatly in screening olive progeny for resistance to leaf spot in breeding programs.