First and fifth authors: University of Georgia, Department of Crop and Soil Sciences, Tifton 31793; second and third authors: U.S. Department of Agriculture-Agricultural Research Service (USDA-ARS), U.S. Horticultural Research Laboratory, Ft. Pierce, FL 34945; fourth author: USDA-ARS, Crop Genetics and Breeding Research Unit, Tifton, GA 31793; and sixth and seventh authors: USDA-ARS, Crop Protection and Management Research Unit, Tifton, GA 31793
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Accepted for publication 13 December 2004.
Late leaf spot disease caused by Cercosporidium personatum is one of the most destructive foliar diseases of peanut (Arachis hypogaea) worldwide. The objective of this research was to identify resistance genes in response to leaf spot disease using microarray and real-time polymerase chain reaction (PCR). To identify transcripts involved in disease resistance, we studied the gene expression profiles in two peanut genotypes, resistant or susceptible to leaf spot disease, using cDNA microarray containing 384 unigenes selected from two expressed sequenced tag (EST) cDNA libraries challenged by abiotic and biotic stresses. A total of 112 spots representing 56 genes in several functional categories were detected as up-regulated genes (log2 ratio > 1). Seventeen of the top 20 genes, each matching gene with known function in GenBank, were selected for validation of their expression levels using real-time PCR. The two peanut genotypes were also used to study the functional analysis of these genes and the possible link of these genes to the disease resistance trait. Microarray technology and real-time PCR were used for comparison of gene expression. The selected genes identified by microarray analysis were validated by real-time PCR. These genes were more greatly expressed in the resistant genotype as a result of response to the challenge of C. personatum than in the susceptible genotype. Further investigations are needed to characterize each of these genes in disease resistance. Gene probes could then be developed for application in breeding programs for marker-assisted selection.
The American Phytopathological Society, 2005