to multiple races of U. appendiculatus, indicating that they are organized in clusters of race-specific genes.Uromyces appendiculatus (Pers.) Unger, the causative fungus of DNA amplification of near isogenic lines (NILs) rust in common beans (Phaseolus vulgaris L.), consists of many patho- Martin et al., 1991) or of contrasting types or pathogenic races. Cultivar Ouro Negro is resistant to most DNA bulks from segregating populations (Michelmore pathotypes detected in Brazil in the last few decades. We attempted et al., 1991) can be used to detect RAPD (Williams et to identify molecular markers linked to a rust resistance gene block present in Ouro Negro. DNA samples extracted from homozygous al., 1990; Welsh and McClelland, 1990) markers in close near isogenic BC 3 F 2:3 lines derived from a cross between susceptible proximity to disease resistance genes. During the develcultivar US Pinto 111 and Ouro Negro were grouped, following greenopment of NILs, these two strategies can be combined house inoculations, into two contrasting bulks, one containing only by constructing DNA bulks from BC n F 2:3 individuals resistant and the other only susceptible plants. The bulks were amplifrom each of two contrasting NILs. This method infied with 605 random primers and two of them amplified bands which creases the chance that markers will be identified close were heteromorphic between the two bulks. These random amplified to the gene of interest (Michelmore et al., 1991; Haley polymorphic DNA (RAPD) bands were transformed into sequence Miklas et al., 1993). characterized amplified regions (SCARs) SCARBA08 and SCARF10. Several RAPD markers associated with genes confer-The rust resistance-susceptibility phenotypes and the molecular genoring resistance to rust in common bean have been identitypes with the two SCAR markers of 303 F 2:3 families from US Pinto fied Haley et al., 1993.; Johnson et 111 ϫ Ouro Negro were determined. SCARBA08 and SCARF10 were determined to be 4.3 Ϯ 1.2 and 6.0 Ϯ 1.3 centimorgans (cM) al., 1995). In some cases, the application of these markfrom the rust resistance locus, respectively. These markers are being ers has been restricted to the laboratories where they used in combination with other markers for resistance to rust, anthracwere identified because of reproducibility problems nose, and angular leaf spot previously identified in our laboratory to
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