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- Identification and validation of microsatellite markers in strawberry tree (Arbutusunedo L.)Publication . Fazenda, Pedro; Pereira, Ricardo; Fonseca, Maria; Carlier, Jorge; Leitão, JoséStrawberry tree (Arbutus unedo L.), an evergreen shrub/small tree of the family Ericaceae, is a main constituent of the Mediterranean basin flora; although it is also found in southwestern Prance, Macaronesia, and Ireland. The small fruits are edible but mostly used for preparation of preserves and jams, and for liquors such as the Portuguese traditional "aguardente de medronho". Traditionally cultivated by small farmers, often in consociation with Quercus sp., strawberry tree is presently emerging as a new important fruit crop cultivated in large orchards by modern export-oriented enterprises. This change of paradigm requires a growing role of plant breeding, upstream of the production process. Genomic tools for this species are mostly limited to the chloroplast genome sequence and to genomic data described in this work. In order to identify strawberry tree microsatellite (SSR) loci we performed partial genome next-generation sequencing using the Ion Torrent technology. The sequenced similar to 24.6M nucleotides resulted in the identification of 1185 microsatellite markers mostly constituted by dinucleotide motifs. The relative amount of microsatellite dinucleotide motifs (AG/CT - 71.7%, AC/GT - 20.5%, AT/AT - 2.9%, and CG/CG - 0.3%) is similar to the one observed in other Ericaceae species. Among a tested sample of 40 SSR primer pairs, 20 amplified well-defined PCR products, 12 (30%) were validated as polymorphic. Used in our collaborative project for molecular identification of selected and improved clones, the identified SSR loci constitute a strong tool for a large panoply of applied and fundamental studies of this emerging fruit crop.
- A microsatellite sequence in the fifth intron provides a broad-spectrum SSR marker for multiple alleles of the er1/PsMLO1 powdery mildew resistance gene in Pisum sativum L.Publication . Cardoso, André; Pereira, Ricardo; Fonseca, Maria; M Leitão, JPowdery mildew caused by the biotrophic ascomycete fungus Erysiphe pisi Syd. is one the most devastating diseases of peas (Pisum sativum L.) with enormous impact in seed production. The most efficient genetic resistance to this disease, so far identified, is conferred by the naturally occurring or experimentally induced by chemical mutagenesis recessive state of the locus er1. Genetically mapped over 2 decades ago, this gene was recently identified as a homolog of the barley (Hordeum sativum L.) powdery mildew resistance gene MLO, and renamed as PsMLO1. The broad wide resistance conferred by the er1/PsMLO1 locus was found to be a consequence of the loss of function of the encoded PsMLO1 protein. After the publication of the expressed sequence of this gene by another research group, we published the genomic sequences of this gene which harbors a relatively long (TA) microsatellite sequence (SSR) in the fifth intron. SSR markers based on this highly polymorphic microsatellite can be used for marker-assisted selection in multiple pea powdery mildew resistance breeding programs involving the er1/PsMLO1 resistance, except in the rare circumstances where the progenitor lines are monomorphic for the microsatellite sequence.