Analyses of expressed sequence tags from the maize foliar pathogen Cercospora zeae-maydis identify novel genes expressed during vegetative, infectious, and reproductive growth

<p>Abstract</p> <p>Background</p> <p>The ascomycete fungus <it>Cercospora zeae-maydis </it>is an aggressive foliar pathogen of maize that causes substantial losses annually throughout the Western Hemisphere. Despite its impact on maize production, little is known about the regulation of pathogenesis in <it>C. zeae-maydis </it>at the molecular level. The objectives of this study were to generate a collection of expressed sequence tags (ESTs) from <it>C. zeae-maydis </it>and evaluate their expression during vegetative, infectious, and reproductive growth.</p> <p>Results</p> <p>A total of 27,551 ESTs was obtained from five cDNA libraries constructed from vegetative and sporulating cultures of <it>C. zeae-maydis</it>. The ESTs, grouped into 4088 clusters and 531 singlets, represented 4619 putative unique genes. Of these, 36% encoded proteins similar (E value ≤ 10^-05) to characterized or annotated proteins from the NCBI non-redundant database representing diverse molecular functions and biological processes based on Gene Ontology (GO) classification. We identified numerous, previously undescribed genes with potential roles in photoreception, pathogenesis, and the regulation of development as well as <it>Zephyr</it>, a novel, actively transcribed transposable element. Differential expression of selected genes was demonstrated by real-time PCR, supporting their proposed roles in vegetative, infectious, and reproductive growth.</p> <p>Conclusion</p> <p>Novel genes that are potentially involved in regulating growth, development, and pathogenesis were identified in <it>C. zeae-maydis</it>, providing specific targets for characterization by molecular genetics and functional genomics. The EST data establish a foundation for future studies in evolutionary and comparative genomics among species of <it>Cercospora </it>and other groups of plant pathogenic fungi.</p

Analyses of expressed sequence tags from the maize foliar pathogen Cercospora zeae-maydis identify novel genes expressed during vegetative, infectious, and reproductive growth

<p>Abstract</p> <p>Background</p> <p>The ascomycete fungus <it>Cercospora zeae-maydis </it>is an aggressive foliar pathogen of maize that causes substantial losses annually throughout the Western Hemisphere. Despite its impact on maize production, little is known about the regulation of pathogenesis in <it>C. zeae-maydis </it>at the molecular level. The objectives of this study were to generate a collection of expressed sequence tags (ESTs) from <it>C. zeae-maydis </it>and evaluate their expression during vegetative, infectious, and reproductive growth.</p> <p>Results</p> <p>A total of 27,551 ESTs was obtained from five cDNA libraries constructed from vegetative and sporulating cultures of <it>C. zeae-maydis</it>. The ESTs, grouped into 4088 clusters and 531 singlets, represented 4619 putative unique genes. Of these, 36% encoded proteins similar (E value ≤ 10^-05) to characterized or annotated proteins from the NCBI non-redundant database representing diverse molecular functions and biological processes based on Gene Ontology (GO) classification. We identified numerous, previously undescribed genes with potential roles in photoreception, pathogenesis, and the regulation of development as well as <it>Zephyr</it>, a novel, actively transcribed transposable element. Differential expression of selected genes was demonstrated by real-time PCR, supporting their proposed roles in vegetative, infectious, and reproductive growth.</p> <p>Conclusion</p> <p>Novel genes that are potentially involved in regulating growth, development, and pathogenesis were identified in <it>C. zeae-maydis</it>, providing specific targets for characterization by molecular genetics and functional genomics. The EST data establish a foundation for future studies in evolutionary and comparative genomics among species of <it>Cercospora </it>and other groups of plant pathogenic fungi.</p

Regulation of Stomatal Tropism and Infection by Light in Cercospora zeae-maydis: Evidence for Coordinated Host/ Pathogen Responses to Photoperiod?

Cercospora zeae-maydis causes gray leaf spot of maize, which has become one of the most widespread and destructive diseases of maize in the world. C. zeae-maydis infects leaves through stomata, which is predicated on the ability of the pathogen to perceive stomata and reorient growth accordingly. In this study, the discovery that light was required for C. zeae-maydis to perceive stomata and infect leaves led to the identification of CRP1, a gene encoding a putative blue-light photoreceptor homologous to White Collar-1 (WC-1) of Neurospora crassa. Disrupting CRP1 via homologous recombination revealed roles in multiple aspects of pathogenesis, including tropism of hyphae to stomata, the formation of appressoria, conidiation, and the biosynthesis of cercosporin. CRP1 was also required for photoreactivation after lethal doses of UV exposure. Intriguingly, putative orthologs of CRP1 are central regulators of circadian clocks in other filamentous fungi, raising the possibility that C. zeae-maydis uses light as a key environmental input to coordinate pathogenesis with maize photoperiodic responses. This study identified a novel molecular mechanism underlying stomatal tropism in a foliar fungal pathogen, provides specific insight into how light regulates pathogenesis in C. zeae-maydis, and establishes a genetic framework for the molecular dissection of infection via stomata and the integration of host and pathogen responses t

Molecular variability in the maize grey leaf spot pathogens in Brazil

Isolates of Cercospora species from leaves displaying symptoms of grey leaf spot were collected in maize-producing areas of south-central Brazil in 2001 and 2002. Restriction digests of the internal transcribed spacer region of rDNA detected the presence of the same two Cercospora species described on maize in the United States, namely C. zeae-maydis and the recently described species, C. zeina . Genetic variability among isolates was assessed by analysing 104 amplified fragment length polymorphism loci. Cluster analysis confirmed the genetic separation of isolates into two species with a mean similarity of 35%. Similarity levels within species were high, averaging 93% and 92% among isolates of C. zeae-maydis and C. zeina , respectively. The mean genetic similarity between C. zeae-maydis and C. zeina and two isolates of C. sorghi f. sp. maydis was 45% and 35%, respectively. Results of this study showed that populations of the grey leaf spot pathogens in Brazil are similar to those in the United States regarding species composition and that C. zeina is also present in Brazil