Phytopathology. 2005 Apr;95(4):381-7. doi: 10.1094/PHYTO-95-0381.
ABSTRACT 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 (log(2) 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.
摘要 由尾孢菌引起的晚疫病是世界范围内花生(落花生)最具破坏性的叶部病害之一。本研究的目的是使用微阵列和实时聚合酶链反应(PCR)来鉴定对叶斑病具有抗性的基因。为了鉴定与抗病性相关的转录本,我们使用 cDNA 微阵列研究了两种花生基因型(对叶斑病具有抗性或易感性)的基因表达谱,该微阵列包含了从两个表达序列标签(EST)cDNA 文库中选择的 384 个基因,这些文库分别受到非生物和生物胁迫的挑战。在几个功能类别中,有 112 个斑点(代表 56 个基因)被检测为上调基因(log2 比值>1)。前 20 个基因中的 17 个,每个基因在 GenBank 中都与已知功能相匹配,被选择用于实时 PCR 验证其表达水平。这两种花生基因型也被用于研究这些基因的功能分析及其与抗病性状的可能联系。微阵列技术和实时 PCR 用于比较基因表达。通过微阵列分析鉴定的选定基因通过实时 PCR 进行了验证。这些基因在对尾孢菌的挑战下,在抗性基因型中的表达水平高于易感基因型。需要进一步研究以表征这些基因在抗病性方面的特性。然后可以开发基因探针用于抗病性的标记辅助选择。