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Quantitative proteomics of Phytophthora infestans life stages

ABSTRACT: Samples were isolated from six different P. infestans life stages (hyphae, sporangia, zoospores, cysts, germinated cysts and appressoria).

INSTRUMENT(S): LTQ Orbitrap XL ETD

ORGANISM(S): Phytophthora Infestans

SUBMITTER: Svante Resjo

LAB HEAD: Svante Resjö

PROVIDER: PXD002446 | Pride | 2017-09-26

REPOSITORIES: Pride

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			Action	DRS
	120113_Svante_01_01x.mgf	Mgf
	120113_Svante_01_01x.mzML.gz	Mzml
	120113_Svante_01_01x.raw	Raw
	120113_Svante_01_01x_rerun.mgf	Mgf
	120113_Svante_01_01x_rerun.mzML.gz	Mzml

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Publications

Proteomic Analysis of Phytophthora infestans Reveals the Importance of Cell Wall Proteins in Pathogenicity.

Resjö Svante S Brus Maja M Ali Ashfaq A Meijer Harold J G HJG Sandin Marianne M Govers Francine F Levander Fredrik F Grenville-Briggs Laura L Andreasson Erik E

Molecular & cellular proteomics : MCP 20170921 11

The oomycete Phytophthora infestans is the most harmful pathogen of potato. It causes the disease late blight, which generates increased yearly costs of up to one billion euro in the EU alone and is difficult to control. We have performed a large-scale quantitative proteomics study of six P. infestans life stages with the aim to identify proteins that change in abundance during development, with a focus on preinfectious life stages. Over 10 000 peptides from 2061 proteins were anal ...[more]

PMID: 28935716

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Profiling the asexual development transcriptome of the late blight pathogen Phytophthora infestans

Project description:Much of the pathogenic success of Phytophthora infestans, the potato and tomato late blight agent, relies on its ability to generate from mycelia large amounts of sporangia, which release zoospores that encyst and form infection structures. To better understand these critical stages, Affymetrix GeneChips based on 15,650 unigenes were designed and used to profile the life cycle, through an analysis of RNA from hyphae, sporangia, cleaving sporangia, motile zoospores, and germinated zoospore cysts. Keywords: Developmental study Gene expression in non-sporulating hyphae, asexual sporangia, sporangia undergoing cleavage, swimming zoospores, and germinated cysts containing appressoria were characterized using isolate 88069. Several strategies controlled for experimental error. Firstly, two replicate hybridizations were used for each tissue type. Secondly, RNAs for the two replicates were prepared by research groups in North America and/or Europe. Thirdly, RNA for each hybridization was pooled from two or three independent cultures made by that group. Finally, before using an RNA sample in an experiment the expression of selected well-characterized genes was measured to ensure that the tissue was properly prepared.

2008-03-14 | E-GEOD-9623 | biostudies-arrayexpress

Profiling the asexual development transcriptome of the late blight pathogen Phytophthora infestans

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Phytophthora have distinct endogenous small RNA populations that include short interfering and microRNAs: Phytophthora infestans small RNA

Project description:Deep sequencing of small RNAs from three Phytophthora species, P. infestans, P. ramorum and P. sojae, was done to systematically analyze small RNA-generating components of Phytophthora genomes. We found that each species produces two distinct small RNA populations that are predominantly 21- or 25-nucleotides long. We present evidence that 25-nucleotide small RNAs are short-interfering RNAs that silence repetitive genetic elements. In contrast, 21-nucleotide small RNAs are associated with inverted repeats, including a novel microRNA family, and may function at the post-transcriptional level. Phytophthora infestans mycelium small RNAs were sequenced and aligned to the P. infestans genome for analysis.

2013-10-21 | E-GEOD-50030 | biostudies-arrayexpress

phytophthora infestans LC/MS/MS

Project description:The oomycete potato blight pathogen Phytophthora infestans secretes a diverse set of proteins to manipulate host plant immunity. However, there is limited knowledge about the role(s) of majority of these secreted proteins. Here we subjected extracellular proteins from in vitro cultured hyphae to LC/MS/MS to investigate the cargo of protein secretion pathways in P. infestans, in particular the conventional secretory pathway. Like the apoplastic effector EPIC1 (a cysteine protease inhibitor), we show that secretion of cell wall degrading enzymes and pathogen associated molecular pattern (PAMP)-like proteins is inhibited by brefeldin A (BFA) in vitro and in planta, demonstrating that these proteins are secreted by the conventional, Golgi-mediated pathway. For comparison, an RXLR (Arg-any amino acid-Leu-Arg) effector, Pi22926, was shown to be unconventionally secreted at haustoria, pathogen structures that penetrate host plant cells, and delivered into infected cells to suppress host defences, consistent with previous findings. We further characterised a pectinesterase (PE), a cell wall degrading enzyme, and INF4, a PAMP-like protein. Both were highly expressed early in infection, enhanced P. infestans colonization, and secreted at haustoria. Notably, in P. infestans transformed lines, PE-mRFP accumulated around the base of haustoria while INF4-mRFP was found to envelop the whole haustorium. We propose that the haustorium is a major site for both non-conventional and conventional secretion, and cell wall degrading enzymes and PAMP proteins are secreted at these sites to facilitate pathogenicity in P. infestans.

2018-09-03 | PXD009802 | Pride

Profiling the secretome and extracellular proteome of the potato late blight pathogen Phytophthora infestans

Project description:Oomycetes are filamentous organisms that cause notorious diseases, several of which with high economic impact. Well known is Phytophthora infestans, the causal agent of potato late blight. Previously, in silico analyses of the genome and transcriptome of P. infestans resulted in the annotation of a large number of genes encoding proteins with an N-terminal signal peptide. This set is collectively referred to as the secretome and comprises proteins involved in, for example, cell wall growth and modification, proteolytic processes and the promotion of successful invasion of plant cells. So far, proteomic profiling in oomycetes was primarily focussed on subcellular, intracellular or cell wall fractions; the extracellular proteome has not been studied systematically. Here we present the first comprehensive characterization of the in vivo secretome and extracellular proteome of P. infestans. We have used mass spectrometry to analyse P. infestans proteins present in seven different growth media with mycelial cultures and this resulted in the consistent identification of over two hundred proteins. Gene ontology classification pinpointed proteins involved in cell wall modifications, pathogenesis, defense responses and proteolytic processes. Moreover, we found members of the RXLR and CRN effector families as well as several proteins lacking an obvious signal peptide. The latter were confirmed to be bona fide extracellular proteins and this suggests that, similar to other organisms, oomycetes exploit non-conventional secretion mechanisms to transfer certain proteins to the extracellular environment.

2016-07-09 | PXD000802 | Pride

Phytophthora infestans miRNA translocation facilitates infection in potato

Project description:Phytophthora spp. encode large sets of effector proteins and distinct populations of small RNAs (sRNAs). Reports suggest that pathogen-derived sRNAs can modulate the expression of plant defense genes. The experiments reported here were designed to shed light on impact of sRNAs in the potato-P. infestans interaction. We used the Argonaute or Ago1 from P. infestans tagged with GFP transformed into the 88069 strain to infect potato cv. Bintje plants. Collected leaf materials were used in co-immunoprecipitation experiments together with P. infestans harboring GFP (control GFP) and P. infestans mycelia grown on media (control mycelia). These three materials were sequenced at a Ion Proton platform. The reads length of 8-38 nt were adaptor-trimmed and mapped to the P. infestans genome and the Solanom tuberosum genome v4.04. Both P. infestans-associated and potato derived sRNAs were identified.

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ChIP-seq of Phytophthora infestans T30-4 mycelium

Project description:Phytophthora infestans, the causal agent of potato late blight, is a devastating plant disease that leads to Irish potato famine and threatens world-wide food security. Despite the genome of P. infestans has provided fundamental resource for studying the aggressiveness of this pandemic pathogen, the epigenomes remain poorly understood. Here, utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS), we demonstrate post-translational modifications (PTM) at P. infestans core histone H3. The PTMs not only include these prevalent modifications in eukaryotes, and also some novel marks, such as H3K53me2 and H3K122me3. We focused on the trimethylations of H3K4, H3K9 and H3K27 and H3K36, and profiled P. infestans epigenomes employing Native Chromatin Immunoprecipitation followed by sequencing (N-ChIP-seq). In parallel, we mapped P. infestans chromomatin accessibility by Assay for Transposase-Accessible Chromatin with high-throughput sequencing (ATAC-seq). We found that adaptive genomic compartments display significantly higher levels of H3K9me3 and H3K27me3, and are generally in condense chromatin. Interestingly, we observed that genes encoding virulence factors, such as effectors, are enriched in open chromatin regions that barely have the four histone modifications. With a combination of genomic, epigenomic, transcriptomic strategies, our study illustrates the epigenetic states in P. infestans, which will help to study genomic functions and regulations in this pathogen.

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Phytophthora infestans

Project description:Phytophthora infestans UK3928

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