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Host-based markers for the early stage of infection by the grapevine trunk pathogen Neofusicoccum parvum

ABSTRACT: In the vineyard, symptoms of the grapevine trunk disease Botryosphaeria dieback do not appear until 1 or 2 years after the causal fungus Neofusicoccum parvum establishes a necrotic canker in the permanent woody structure of the vine. There are preventative management practices, but growers tend to wait until symptoms are widespread, at which point prevention has limited efficacy. Toward development of an early detection tool, we examined the leaves of inoculated vs. non-inoculated plants for differential gene expression via RNASeq. Stems were examined to monitor spread of the infection, and its spatial and temporal relationship to anatomical changes via light microscopy and high resolution computed tomography (HRCT). The early stage of infection occurred prior to 2 months post-inoculation (MPI), at which point spread of the pathogen beyond the inoculation site was greatest. This incubation period was also characterized by the largest stem lesions, the highest levels of fungal colonization and xylem vessels fully-occluded by gels, and the lowest starch content of xylem fibers and rays. Prior to 2 MPI, RNASeq and validative qPCR analysis identified eight candidate markers, which are transcriptionally activated by infection, but not by wounding alone. Our best marker genes, a dehydrin, a BURP domain containing protein and an abscissic acid-induced wheat plasma membrane polypeptide 19 (AWPM-19), identified the pathogen’s presence with high specificity. Screening of genome-wide expression data revealed that this signature is not affected by many abiotic and biotic stresses.

ORGANISM(S): Vitis vinifera

PROVIDER: GSE58653 | GEO | 2015/03/24

SECONDARY ACCESSION(S): PRJNA253108

REPOSITORIES: GEO

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Host-based markers for the early stage of infection by the grapevine trunk pathogen Neofusicoccum parvum

Project description:In the vineyard, symptoms of the grapevine trunk disease Botryosphaeria dieback do not appear until 1 or 2 years after the causal fungus Neofusicoccum parvum establishes a necrotic canker in the permanent woody structure of the vine. There are preventative management practices, but growers tend to wait until symptoms are widespread, at which point prevention has limited efficacy. Toward development of an early detection tool, we examined the leaves of inoculated vs. non-inoculated plants for differential gene expression via RNASeq. Stems were examined to monitor spread of the infection, and its spatial and temporal relationship to anatomical changes via light microscopy and high resolution computed tomography (HRCT). The early stage of infection occurred prior to 2 months post-inoculation (MPI), at which point spread of the pathogen beyond the inoculation site was greatest. This incubation period was also characterized by the largest stem lesions, the highest levels of fungal colonization and xylem vessels fully-occluded by gels, and the lowest starch content of xylem fibers and rays. Prior to 2 MPI, RNASeq and validative qPCR analysis identified eight candidate markers, which are transcriptionally activated by infection, but not by wounding alone. Our best marker genes, a dehydrin, a BURP domain containing protein and an abscissic acid-induced wheat plasma membrane polypeptide 19 (AWPM-19), identified the pathogenâ??s presence with high specificity. Screening of genome-wide expression data revealed that this signature is not affected by many abiotic and biotic stresses. Asymptomatic leaf mRNA profiles of Neofusicoccum parvum infected wounded plants (IW) and non-infected wounded plants (NIW) at two different time points post inoculation (0 MPI, 0.5-1.5 MPI) were generated by deep sequencing in biological triplicates using Illumina HiSeq2500 technology.

2015-03-24 | E-GEOD-58653 | biostudies-arrayexpress

Aggressive Potato virus YNTN (PVYNTN) and mild PVYN isolates trigger different specific responses in susceptible potato

Project description:Potato virus YNTN (PVYNTN) is one of the most devastating potato virus causing great losses in the potato production industry. PVYNTN induces severe symptoms on inoculated leaves and a disease known as potato tuber necrosis ringspot disease (PTNRD) develops on tubers. Closely related PVYN isolate induces only mild symptoms on inoculated potato leaves and no symptoms on tubers. The early response of sensitive potato cvs. Igor and Nadine to inoculation with PVYNTN and PVYN was analysed allowing identification of genes involved in severe symptoms induction. Microarray and quantitative-PCR analysis was carried out to identify differentially expressed genes after inoculation with both virus isolates. Two distinct groups of genes were shown to have a role in severe symptoms development – one group of genes related to energy production and a second group of genes connected with virus spread. Earlier accumulation of sugars and decrease in photosynthesis was observed in leaves inoculated with aggressive PVYNTN isolate than in leaves inoculated with milder PVYN isolate. PVYNTN isolate was shown not to activate differential expression of antioxidant metabolism and pectinmethylesterase inhibitor (PMEI) leading to a delay in plant response and on the other hand it limited callose deposition enabling faster virus spread through the plant.

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Project description:Potato virus YNTN (PVYNTN) is one of the most devastating potato virus causing great losses in the potato production industry. PVYNTN induces severe symptoms on inoculated leaves and a disease known as potato tuber necrosis ringspot disease (PTNRD) develops on tubers. Closely related PVYN isolate induces only mild symptoms on inoculated potato leaves and no symptoms on tubers. The early response of sensitive potato cvs. Igor and Nadine to inoculation with PVYNTN and PVYN was analysed allowing identification of genes involved in severe symptoms induction. Microarray and quantitative-PCR analysis was carried out to identify differentially expressed genes after inoculation with both virus isolates. Two distinct groups of genes were shown to have a role in severe symptoms development â one group of genes related to energy production and a second group of genes connected with virus spread. Earlier accumulation of sugars and decrease in photosynthesis was observed in leaves inoculated with aggressive PVYNTN isolate than in leaves inoculated with milder PVYN isolate. PVYNTN isolate was shown not to activate differential expression of antioxidant metabolism and pectinmethylesterase inhibitor (PMEI) leading to a delay in plant response and on the other hand it limited callose deposition enabling faster virus spread through the plant. Each microarray was hybridized with PVYNTN inoculated sample and PVYN inoculated sample from the same biological replicate. Three biological replicates were analyzed.

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2013-11-16 | E-GEOD-52428 | biostudies-arrayexpress

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Project description:Potato virus YNTN (PVYNTN), causing potato tuber ring necrosis disease, dramatically lowers the quantity and the quality of the potato yield all over the world. The cultivar Igor is one of the most susceptible cultivars, developing severe disease symptoms on plants as well as on tubers. Finding genes differentially expressed in the early response to infection, when the host response is more defense- than infection- related, could improve our understanding of the potato - PVYNTN interaction. Differential gene expression in early response of potato cv. Igor plants to PVYNTN infection was studied using potato TIGR cDNA-microarrays. Expression was compared between mock inoculated and virus infected plants 12 hours after inoculation, in four biological replicates. Keywords: direct comparison Each microarray was hybridized with a virus inoculated sample and mock inoculated sample from the same biological replicate. Four biological replicates were analyzed.

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