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Tomato xylem proteome composition following infection by Fusarium oxysporum

ABSTRACT: The influence of during colonization by Fusarium oxysporum f. sp. Lycopersici secreted effector proteins on the proteome of the xylem sap of tomato plants was investigated using a label-free quantitative proteomics approach. A comparison was made between plants inoculated with either a mock control, a non-effector knockout control, Fusarium oxysporum Fol007 wildtype and four Fol007 single effector protein knockout strains. Specific effects on the relative abundance of certain proteins of the xylem sap occurred for the different knockout strains next to a core set of 24 differentially accumulated proteins which may provide insights into the mechanisms of promoting infection for each of the tested effector proteins.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Solanum Lycopersicum Fusarium Oxysporum F. Sp. Lycopersici

TISSUE(S): Xylem

SUBMITTER: Sjef Boeren

LAB HEAD: Sjef Boeren

PROVIDER: PXD003010 | Pride | 2015-10-26

REPOSITORIES: Pride

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			Action	DRS
	FT10_infected_d65.RAW	Raw
	FT11_infected_d65.RAW	Raw
	FT12_infected_d65.RAW	Raw
	FT13_infected_with_07.RAW	Raw
	FT14_infected_with_07.RAW	Raw

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Publications

The effector repertoire of Fusarium oxysporum determines the tomato xylem proteome composition following infection.

Gawehns Fleur F Ma Lisong L Bruning Oskar O Houterman Petra M PM Boeren Sjef S Cornelissen Ben J C BJ Rep Martijn M Takken Frank L W FL

Frontiers in plant science 20151104

Plant pathogens secrete small proteins, of which some are effectors that promote infection. During colonization of the tomato xylem vessels the fungus Fusarium oxysporum f.sp. lycopersici (Fol) secretes small proteins that are referred to as SIX (Secreted In Xylem) proteins. Of these, Six1 (Avr3), Six3 (Avr2), Six5, and Six6 are required for full virulence, denoting them as effectors. To investigate their activities in the plant, the xylem sap proteome of plants inoculated with Fol wild-type or ...[more]

PMID: 26583031

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Changes in the Proteome of Xylem Sap in Brassica oleracea in Response to Fusarium oxysporum Stress

Project description:Fusarium oxysporum f. sp. conlutinans (Foc) is a serious root-invading and xylem-colonizing fungus that causes yellowing in Brassica oleracea. To comprehensively understand the interaction between F. oxysporum and B. oleracea, composition of the xylem sap proteome of the non-infected and Foc-infected plants was investigated in both resistant and susceptible cultivars using liquid chromatography-tandem mass spectrometry (LC-MS/MS) after in-solution digestion of xylem sap proteins. Whole genome sequencing of Foc was carried out and generated a predicted Foc protein database. The predicted Foc protein database was then combined with the public B. oleracea and B. rapa protein databases downloaded from Uniprot and used for protein identification. About 200 plant proteins were identified in the xylem sap of susceptible and resistant plants. Comparison between the non-infected and Foc-infected samples revealed that Foc infection causes changes to the protein composition in B. oleracea xylem sap where repressed proteins accounted for a greater proportion than those of induced in both the susceptible and resistant reactions. The analysis on the proteins with concentration change >=2 fold indicated a large portion of up- and down-regulated proteins were those acting on carbohydrates. Proteins with leucine-rich repeats and legume lectin domains were mainly induced in both resistant and susceptible system, so was the case of thaumatins. Twenty-five Foc proteins were identified in the infected xylem sap and ten of them were cysteine-containing secreted small proteins that are good candidates for virulence and/or avirulence effectors. The findings of differential response of protein contents in the xylem sap between the non-infected and Foc-infected samples as well as the Foc candidate effectors secreted in xylem provide valuable insights into B. oleracea-Foc interactions.

2016-01-26 | PXD003378 | Pride

Tomato xylem sap proteome composition following endophyte- and R-gene mediated resistance towards Fusarium oxysporum f.sp. lycopersici inoculation.

Project description:Xylem sap proteome studies on susceptible or resistant tomato (Solanum lycopersicum) inoculated with endophytic and/or pathogenic strains of Fusarium oxysporum f.sp. lycopersici were conducted to get insights into the molecular differences between endophyte- and R-gene-mediated resistance (EMR and RMR). The EMR and RMR proteomes were compared to each other and to the mock control. Interestingly, specific PR-5 isoforms were found to exclusively accumulate during endophyte or genetic resistance, providing excellent markers to distinguish both resistance types at the molecular level.

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Analyses of the Xylem Sap Proteomes Identified Candidate Fusarium virguliforme Proteinacious Toxins

Project description:Sudden death syndrome (SDS) is caused by the ascomycete fungus, Fusarium virguliforme. The pathogen secretes one or more toxins into the infected soybean roots to cause foliar SDS. This study investigated if the xylem sap of F. virguliforme-infected soybean plants contains secreted F. virguliforme-proteins, some of which could be involved in foliar SDS development.

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Microarray expression analysis of transgenic Arabidopsis thaliana

Project description:Comparison of expression differences between Col-0 Arabidopsis thaliana and transgenic plants in the same background carrying three different Fusarium oxysporum effector genes

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Multi‐omics analysis of xylem sap uncovers dynamic modulation of poplar defenses by ammonium and nitrate

Project description:Xylem sap is the major transport route for nutrients from roots to shoots. In the present study, we investigated how variations in nitrogen (N) nutrition affected the metabolome and proteome of xylem sap and the growth of the xylem endophyte Brennaria salicis, and we also report transcriptional re-wiring of leaf defenses in poplar (Populus × canescens). We supplied poplars with high, intermediate or low concentrations of ammonium or nitrate. We identified 288 unique proteins in xylem sap. Approximately 85% of the xylem sap proteins were shared among ammonium- and nitrate-supplied plants. The number of proteins increased with increasing N supply but the major functional categories (catabolic processes, cell wall-related enzymes, defense) were unaffected. Ammonium nutrition caused higher abundances of amino acids and carbohydrates, whereas nitrate caused higher malate levels in xylem sap. Pipecolic acid and N-hydroxy-pipecolic acid increased, whereas salicylic acid and jasmonoyl-isoleucine decreased, with increasing N nutrition. Untargeted metabolome analyses revealed 2179 features in xylem sap, of which 863 were differentially affected by N treatments. We identified 124 metabolites, mainly from specialized metabolism of the groups of salicinoids, phenylpropanoids, phenolics, flavonoids, and benzoates. Their abundances increased with decreasing N, except coumarins. Brennaria salicis growth was reduced in nutrient-supplemented xylem sap of low- and high- NO3 - -fed plants compared to that of NH4 + -fed plants. The drastic changes in xylem sap composition caused massive changes in the transcriptional landscape of leaves and recruited defenses related to systemic acquired and induced systemic resistance. Our study uncovers unexpected complexity and variability of xylem composition with consequences for plant defenses.

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Expression profiling of Fusarium oxysporum infected wild-type Arabidopsis and a 35S Fusarium SIX4 overexpression line.

2016-05-31 | GSE75928 | GEO

Expression profiling of Fusarium oxysporum infected wild-type Arabidopsis and a 35S Fusarium SIX4 overexpression line.

Project description:Fungal effectors play important roles in inciting disease development on host plants. We identified an effector (Secreted in Xylem4, SIX4) in an Arabidopsis infecting isolate (Fo5176) of the root-infecting fungal pathogen Fusarium oxysporum and demonstrated this effector is required for full virulence. To explore the role of Fo5176_SIX4 we use whole transcriptome profiling of root tissues from plants overexpressing this effector (35sSIX4) versus wild-type (Col-0) plants after F. oxysporum infection. We grew both WT and 35sSIX4 plants for four weeks in soil. After four weeks the plants were infected with Fusarium oxyporum isolate Fo5176, trays covered with a plastic dome and incubated at 28C. There were four independent replicates of each treatment and each replicate contained root tissue from 20 plants. Each replicate (8 in total) was harvested 4 days post inoculation and the resulting RNA was used for hybridization to an Affymetrix ATH1 chip.

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A new strategy to identify the long-distance mobile peptides from xylem sap.

Project description:Total RNAs were extracted from internodes and roots of 18 day-after-germination soybean plants, and the expression levels of the genes that encode xylem sap associated peptides or proteins were analyzed.

2015-10-23 | E-MTAB-5052 | biostudies-arrayexpress

Xylem sap shotgun proteomics from Mn-excess tomato plants

Project description:The aim of this work was to study the effects of Mn excess on the protein profile of the xylem sap of tomato (Solanum lycopersicum), with the aim of elucidating plant response mechanisms to these nutritional stresses. Tomato was chosen as a model plant because the tomato genome has been published and this plant species has adequate root pressure and turgid stems that permit xylem sap sampling in sufficient amounts by de-topping. The high- throughput shotgun analysis has permitted to identify and quantitate a large number of low abundance proteins in the tomato xylem sap.

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