454 Genome sequencing of Pseudoperonospora cubensis reveals effector proteins with a QXLR translocation motif.
Pseudoperonospora cubensis is a biotrophic oomycete pathogen that causes downy mildew of cucurbits, a devastating foliar disease threatening cucurbit production worldwide. We sequenced P. cubensis genomic DNA using 454 pyrosequencing and obtained random genomic sequences covering approximately 14% of the genome, thus providing the first set of useful genomic sequence information for P. cubensis. Using bioinformatics approaches, we identified 32 putative RXLR effector proteins. Interestingly, we also identified 29 secreted peptides with high similarity to RXLR effectors at the N-terminal translocation domain, yet containing an R-to-Q substitution in the first residue of the translocation motif. Among these, a family of QXLR-containing proteins, designated as PcQNE, was confirmed to have a functional signal peptide and was further characterized as being localized in the plant nucleus. Internalization of secreted PcQNE into plant cells requires the QXLR-EER motif. This family has a large number of near-identical copies within the P. cubensis genome, is under diversifying selection at the C-terminal domain, and is upregulated during infection of plants, all of which are common characteristics of characterized oomycete effectors. Taken together, the data suggest that PcQNE are bona fide effector proteins with a QXLR translocation motif, and QXLR effectors are prevalent in P. cubensis. Furthermore, the massive duplication of PcQNE suggests that they might play pivotal roles in pathogen fitness and pathogenicity.
Department of Plant Pathology, Michigan State University, East Lansing, MI 48824, USA.
Miaoying
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Tian
Joe
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Win
Elizabeth
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Savory
Alyssa
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Burkhardt
Michael
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Held
Federica
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Brandizzi
Brad
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D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019295:Computational Biology; D018553:Cucumis sativus / Q000382:microbiology; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016681:Genome, Fungal; D008969:Molecular Sequence Data; D009868:Oomycetes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D021382:Protein Sorting Signals; D021381:Protein Transport; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000379:methods*; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000378:metabolism* / Q000382:microbiology
D004271:DNA, Fungal; D005656:Fungal Proteins; D021382:Protein Sorting Signals