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Inheritance and Characterization of Angular Leaf Spot Resistance Gene Present in Common Bean Accession G 10474 and Identification of an AFLP Marker Linked to the Resistance Gene

Centro Internacional de Agricultura Tropical, A. A. 6713, Cali, Colombia

View larger version (69K): [in a new window]   Fig. 1. AFLP banding patterns from the combination E-ACA/M-CTT330 for the cross G 10474 x Sprite. The polymorphic fragments are indicated by the arrow. MP = molecular marker, RP is resistant parent (G 10474), RB is resistant bulk, SB is susceptible bulk, Lanes 1 to 5 are resistant plants while Lanes 6 to 10 are susceptible F2 plants, SP is the susceptible parent (Sprite).

View larger version (63K): [in a new window]   Fig. 2. PCR products from F2 population showing homozygous resistant plants (305 bp), homozygous susceptible plants (280 bp) and heterozygous plants with both fragments. The amplification products were obtained using the SCAR marker derived from the polymorphic AFLP fragment generated by the primer combination E-ACA/M-CTT330. Lane 19 is negative control (no plant DNA) while 20 is the 100-bp molecular size marker.

View larger version (65K): [in a new window]   Fig. 3. PCR amplification of common bean angular leaf spot differential genotypes using the SCAR marker derived from the polymorphic AFLP fragment generated by the primer combination E-ACA/M-CTT330. Lanes 1–6 are Andean common bean genotypes, Lane 7 is resistant parent G 10474, Lane 8 = resistant bulk, Lane 9 = susceptible bulk, Lane 10 = susceptible parent, Sprite, while Lanes 11-16 are Mesoamerican ALS differential bean genotypes, including Mexico 54 in Lane 14. Lane 17 is negative control while MP is the 100-bp molecular size marker.

View larger version (67K): [in a new window]   Fig. 4. Effect of different DNA extraction methods on detection of SCAR marker linked to the ALS resistance locus of G 10474. DNA was extracted using the alkaline method, ammonium acetate method, and the phenol/chloroform method as described in Materials and Methods. Lane 1 to 3 are Andean common bean genotypes Don Timoteo, G 11796 and Bolon Bayo, while Lanes 4 to 6 are Mesoamerican genotypes G 10474, Mexico 54 and Flor de mayo, respectively. Lane c = negative control (no plant DNA added) while lane MP is the 100-bp molecular size marker.