Identification of RAPD markers for common root rot and spot blotch (Cochliobolus sativus) resistance in barley.

Abstract

The identification of RAPD markers associated with genes for resistance to Cochliobolus sativus in barley would increase the efficiency of gene manipulation by reducing the number of lines that must be evaluated from a resistant by susceptible cross and by allowing selection during the off season. Two barley crosses consisting of resistant and susceptible parent genotypes ('Virden' x 'Ellice' and Fr926-77 x 'Deuce', both 2 row x 6 row crosses) and more than 140 homozygous progeny lines were rated for their reactions in field nurseries to common root rot and in a controlled environment for spot blotch. Putative RAPD markers were identified using bulked segregant analysis followed by individual progeny line analyses. Polymorphisms associated with disease reaction were detected between bulked segregant samples as differences in the band intensity of DNA fragments. The bulked segregant samples were screened against 186 RAPD primers (decamers) using the polymerase chain reaction. For the cross Fr926-77 x 'Deuce', one RAPD marker was obtained that did not segregate as expected but was associated with both diseases. For the cross 'Virden' x 'Ellice', a single RAPD marker was obtained that did not have the expected segregation ratio but was associated with spot blotch reaction. One RAPD marker linked to 2-rowed and 6-rowed spike locus was obtained in each cross, and both the marker and row type were associated with common root rot and spot blotch reactions. For the cross 'Virden' x 'Ellice', a linkage group consisting of three RAPD markers was associated with common root rot and spot blotch reaction. The genes associated with these markers condition significant levels of resistance to C. sativus and may be used to increase the speed and precision of resistance gene manipulation in barley germplasm.