An EST library from Puccinia graminis f. sp. tritici reveals genes potentially involved in fungal differentiation

Broeker, Katja; Frank, Bernard; Moerschbacher, Bruno M.

doi:10.1111/j.1574-6968.2006.00127.x

Cited by 27 publications

(21 citation statements)

References 30 publications

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“…Proteins involved in protein synthesis, primary metabolism and energy production were most prevalent in predicted proteins with known functions. Similar findings were also reported in other pathogenic interactions during infection [19,22,24]. Overall, the less redundant sequences were surprisingly unique, especially considering the database searches included nucleotide and predicted protein searches of the P. graminis f. sp.…”

Section: Discussionsupporting

confidence: 81%

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Generation and analysis of expression sequence tags from haustoria of the wheat stripe rust fungus Puccinia striiformis f. sp. Tritici

et al. 2009

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BackgroundStripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most destructive diseases of wheat (Triticum aestivum L.) worldwide. In spite of its agricultural importance, the genomics and genetics of the pathogen are poorly characterized. Pst transcripts from urediniospores and germinated urediniospores have been examined previously, but little is known about genes expressed during host infection. Some genes involved in virulence in other rust fungi have been found to be specifically expressed in haustoria. Therefore, the objective of this study was to generate a cDNA library to characterize genes expressed in haustoria of Pst.ResultsA total of 5,126 EST sequences of high quality were generated from haustoria of Pst, from which 287 contigs and 847 singletons were derived. Approximately 10% and 26% of the 1,134 unique sequences were homologous to proteins with known functions and hypothetical proteins, respectively. The remaining 64% of the unique sequences had no significant similarities in GenBank. Fifteen genes were predicted to be proteins secreted from Pst haustoria. Analysis of ten genes, including six secreted protein genes, using quantitative RT-PCR revealed changes in transcript levels in different developmental and infection stages of the pathogen.ConclusionsThe haustorial cDNA library was useful in identifying genes of the stripe rust fungus expressed during the infection process. From the library, we identified 15 genes encoding putative secreted proteins and six genes induced during the infection process. These genes are candidates for further studies to determine their functions in wheat-Pst interactions.

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Section: Discussionsupporting

confidence: 81%

“…The frequency of redundant ESTs was shown in Figure 1. The average G+C content of these unique sequences was 43.11%, which was similar to the G+C content of ESTs in P. graminis and Pst germinated urediniospores [17,19]. The sequences were deposited in the NCBI dbEST sequence database (Accession numbers GH737012 - GH738498).…”

Section: Resultsmentioning

confidence: 67%

Generation and analysis of expression sequence tags from haustoria of the wheat stripe rust fungus Puccinia striiformis f. sp. Tritici

et al. 2009

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“…The transcript 00302 is an ortholog of the M. oryzae P-type ATPase gene ( pde1 ), required for the development of penetration hyphae and the proliferation of the fungus (Balhadère and Talbot, 2001) and was detected in the three H. vastatrix libraries at relatively constant expression levels. Orthologs of this gene were also identified in other rusts species (Broeker et al, 2006; Jakupović et al, 2006; Yin et al, 2009; Duplessis et al, 2011a), with elevated expression values in P. pachyrhizi appressoria (Stone et al, 2012). Among members of the Voltage-gated K + Channel β subunit family are two transcripts (00184 and 00427) identified in the three H. vastatrix libraries, and one (04218) only identified in the H library but at higher Ra values.…”

Section: Resultsmentioning

confidence: 84%

Overview of the functional virulent genome of the coffee leaf rust pathogen Hemileia vastatrix with an emphasis on early stages of infection

et al. 2014

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Hemileia vastatrix is the causal agent of coffee leaf rust, the most important disease of coffee Arabica. In this work, a 454-pyrosequencing transcriptome analysis of H. vastatrix germinating urediniospores (gU) and appressoria (Ap) was performed and compared to previously published in planta haustoria-rich (H) data. A total of 9234 transcripts were identified and annotated. Ca. 50% of these transcripts showed no significant homology to international databases. Only 784 sequences were shared by the three conditions, and 75% were exclusive of either gU (2146), Ap (1479) or H (3270). Relative transcript abundance and RT-qPCR analyses for a selection of genes indicated a particularly active metabolism, translational activity and production of new structures in the appressoria and intense signaling, transport, secretory activity and cellular multiplication in the germinating urediniospores, suggesting the onset of a plant-fungus dialogue as early as at the germ tube stage. Gene expression related to the production of carbohydrate-active enzymes and accumulation of glycerol in germinating urediniospores and appressoria suggests that combined lytic and physical mechanisms are involved in appressoria-mediated penetration. Besides contributing to the characterization of molecular processes leading to appressoria-mediated infection by rust fungi, these results point toward the identification of new H. vastatrix candidate virulence factors, with 516 genes predicted to encode secreted proteins.

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“…Low-level representation of fungal genes may be because they harvested infected tissues at early time points. Suppression subtractive hybridization was also applied to identify genes expressed during the pathogen's interaction with its host [20,21]. Broeker et al [21] identified 81 Puccinia graminis f. sp.…”

Section: Introductionmentioning

confidence: 99%

“…Suppression subtractive hybridization was also applied to identify genes expressed during the pathogen's interaction with its host [20,21]. Broeker et al [21] identified 81 Puccinia graminis f. sp. tritici genes by sequencing 454 random clones from rust-infected wheat plants subtracted from those of healthy leaves.…”

Section: Introductionmentioning

confidence: 99%

Identification and analysis of in planta expressed genes of Magnaporthe oryzae

Park

et al. 2010

BMC Genomics

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BackgroundInfection of plants by pathogens and the subsequent disease development involves substantial changes in the biochemistry and physiology of both partners. Analysis of genes that are expressed during these interactions represents a powerful strategy to obtain insights into the molecular events underlying these changes. We have employed expressed sequence tag (EST) analysis to identify rice genes involved in defense responses against infection by the blast fungus Magnaporthe oryzae and fungal genes involved in infectious growth within the host during a compatible interaction.ResultsA cDNA library was constructed with RNA from rice leaves (Oryza sativa cv. Hwacheong) infected with M. oryzae strain KJ201. To enrich for fungal genes, subtraction library using PCR-based suppression subtractive hybridization was constructed with RNA from infected rice leaves as a tester and that from uninfected rice leaves as the driver. A total of 4,148 clones from two libraries were sequenced to generate 2,302 non-redundant ESTs. Of these, 712 and 1,562 ESTs could be identified to encode fungal and rice genes, respectively. To predict gene function, Gene Ontology (GO) analysis was applied, with 31% and 32% of rice and fungal ESTs being assigned to GO terms, respectively. One hundred uniESTs were found to be specific to fungal infection EST. More than 80 full-length fungal cDNA sequences were used to validate ab initio annotated gene model of M. oryzae genome sequence.ConclusionThis study shows the power of ESTs to refine genome annotation and functional characterization. Results of this work have advanced our understanding of the molecular mechanisms underpinning fungal-plant interactions and formed the basis for new hypothesis.

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An EST library from Puccinia graminis f. sp. tritici reveals genes potentially involved in fungal differentiation

Cited by 27 publications

References 30 publications

Generation and analysis of expression sequence tags from haustoria of the wheat stripe rust fungus Puccinia striiformis f. sp. Tritici

Generation and analysis of expression sequence tags from haustoria of the wheat stripe rust fungus Puccinia striiformis f. sp. Tritici

Overview of the functional virulent genome of the coffee leaf rust pathogen Hemileia vastatrix with an emphasis on early stages of infection

Identification and analysis of in planta expressed genes of Magnaporthe oryzae

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