Molecular characterisation and polymorphism of MinLm1, a minisatellite from the phytopathogenic ascomycete Leptosphaeria maculans

A sequence-characterised amplified region marker was identified in the phytopathogenic fungus Leptosphaeria maculans, which generated a single-banding pattern corresponding to six alleles showing size polymorphism between L. maculans field isolates. The size polymorphism was due to 2–7 tandem repeats of the 23-bp motif 5′ TCTTACTTACATACACACCTCCC 3′. The repeated sequence, termed MinLm1, shares many features specific to minisatellites, e.g. a very strong G/C strand asymmetry, the presence of 6-bp direct repeats at both ends of the sequence and its occurrence in a region rich in microsatellites such as (CT)n, (ATG)n, (GTG)n and (CAT)n. MinLm1 shows a very high degree of conservation of the bases from one repeat to another and from one isolate to another (percent match range: 99.6–100%), whatever their geographical or temporal relatedness. MinLm1is a single-locus minisatellite located on chromosomes sized 2.79 Mb and 2.48 Mb, of L. maculansisolates a.2 and H5, respectively. In agricultural populations of L. maculans, two alleles of MinLm1 were prevalent, corresponding to 2× and 5× repeats of the core motif. Differences in allele frequencies were observed in some cropping conditions, suggesting that MinLm1is an informative marker for epidemiological studies of the pathogen.