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Analysis of transcriptional response to mutated Potyvirus Potato virus A that induces necrotic symptoms.

ABSTRACT: Mutations introduced to a highly variable region in HCpro reduced HCpro-HIP2 interactions and virulence of PVA, and induced novel types of necrotic symptoms in the systemically infected leaves of Nicotiana species. Microarray-based analysis of gene expression revealed induction of large numbers of defence-related genes including jasmonic acid and ethylene inducible pathways in the leaves systemically infected by the mutated PVA.

ORGANISM(S): Nicotiana tabacum

PROVIDER: GSE42833 | GEO | 2013/03/23

SECONDARY ACCESSION(S): PRJNA183700

REPOSITORIES: GEO

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Analysis of transcriptional response to mutated Potyvirus Potato virus A that induces necrotic symptoms.

Project description:Mutations introduced to a highly variable region in HCpro reduced HCpro-HIP2 interactions and virulence of PVA, and induced novel types of necrotic symptoms in the systemically infected leaves of Nicotiana species. Microarray-based analysis of gene expression revealed induction of large numbers of defence-related genes including jasmonic acid and ethylene inducible pathways in the leaves systemically infected by the mutated PVA. Three treaments: infection with wild type PVA (WT), infection with mutated PVA (MUT), and mock-inoculation (Mock). Six plants were treated for each treatment. Experiment was repeated three times. WT and MUT samples from all three experiments were hybridised on microarray. Mock samples from experiments 1 and 2 were hybridised as healthy controls.

2013-03-23 | E-GEOD-42833 | biostudies-arrayexpress

Microarray transcription profiling of healthy and PVYNTN infected potato tuber tissues (necrotic and non-necrotic)

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Local and systemic gene expression in potato infected with PVY

Project description:The goal of these studies is analysis of gene expression profiles in potato plants infected with potato virus Y (necrotic strain - PVYN). A prototypic plant pathogen interaction system used to study disease resistance is tobacco (Nicotiana tabacum) and tobacco mosaic virus (TMV). In tobacco, the resistance response comprises two phenomena, known as the hypersensitive response (HR) and systemic acquired resistance (SAR). HR is typically characterized by a local necrosis surrounding the site of viral entry leading to restriction of pathogen replication and spread. SAR is the enhanced ability of the plant to resist a secondary challenge by the same or a different pathogen inoculated elsewhere on the plant. The identity of the signal(s) that induces HR and SAR is still poorly understood. Plants of S. tuberosum cv. Rywal, resistant to all known (N, 0, NTN) PVY strains, were grown in growth chambers using 16 h period of light (22°C) and 8 h of darkness (18°C). For all experiments, 6-week-old plants were used. For inoculation with PVY, carborundum-dusted leaves were rubbed with water (mock control) or PVYN solution (1 µg ml-1). Two to four leaves were inoculated on each plant and harvested together at 1, 3, and 6 days post inoculation (dpi). Separately, noninoculated leaves were harvested at the same time points (upper leaves). The first time point (1 dpi) has been chosen to study the early stage of plant response before necrotic lesion formation. At 3 dpi necrotic lesions start to appear, and at 6 dpi necroses are fully developed which correlates with the maximum of salicylic acid increase in virus inoculated leaves. Keywords: Direct comparison

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Potato virus Y HCPro suppression of antiviral silencing in Nicotiana benthamiana plants correlates with its ability to bind in vivo to small RNAs of 21 and 22 nucleotides in length of viral sequence

Project description:We have investigated short and small RNAs (sRNAs) that were bound to a biologically active hexahistidine-tagged Potato virus Y (PVY) HCPro suppressor of silencing, expressed from a heterologous virus vector in Nicotiana benthamiana plants, and purified under non-denaturing conditions. We found that RNAs in purified preparations were differentially enriched in sRNAs of 21 and to a much lesser extent of 22 nucleotides (nt) in length and of viral sequence (vsRNAs) when compared to those found in a control plant protein background bound to the nickel resin in the absence of HCPro, or in a purified alanine substitution HCPro mutant (HCPro mutB) control that lacked suppressor of silencing activity. In both controls, sRNAs were composed almost entirely by molecules of plant sequence, indicating that the resin-bound protein background had no affinity for vsRNAs, and also that HCPro mutB failed to bind to vsRNAs. Therefore, PVY HCPro suppressor activity correlated with its ability to bind to 21 and 22 nt vsRNAs. HCPro constituted at least 54 % of the total protein content in purified preparations and we were able to calculate its contribution to the 21 and the 22 nt pool of sRNAs present in the purified samples and its binding strength relative to the background. We also found that in the 21 nt vsRNAs of the HCPro preparation 5´ terminal adenines were overrepresented relative to the controls, but not in vsRNAs of other sizes or of plant sequence.

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2006-03-23 | E-MEXP-290 | biostudies-arrayexpress

Effect of two potato viruses, PLRV and PVY, on potato gene expression in Russet Burbank

Project description:Potato plants (cv. Russet Burbank) were grown as in vitro plantlets, and transplanted into 12-cm pots in Sunshine Mix #2 (peat/perlite mix) supplemented with slow release fertilizer (Osmocoat) in the greenhouse. The experiment was conducted in the summer of 2004 at Moscow, ID. Plantings were staggered to facilitate collection of volatiles and leaf samples during a short harvest window for all times of all treatments. Infectious Green Peach Aphids were raised on PLRV-infected potato plants; test plants were infected by placing 10 infectious aphids on a leaf in a clip cage for 48 h. The control plants for PLRV were similarly treated with aphids that had been raised on uninfected potato plants. A local isolate of PVYO was inoculated mechanically using purified virus (5 ìg ml-1) in buffer (50 mM Na2HPO4, 20 mM Na2SO3 pH 7.0) lightly rubbed onto a leaf previously dusted with carborundum. Control plants for PVY were treated in the same manner using buffer without PVY. The RNA samples were made from systemically affected leaves, excluding the actual leaves inoculated by aphids or that received the mechanical infection. Leaf volatiles were concurrently collected from equivalent plants; aliquots of frozen leaf samples were sent for metabolite analysis prior to RNA extraction. Leaf harvests were made 1, 3, 7, 14, and 28 d post-inoculation. Keywords: Direct comparison

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LC-MSMS of RIBOPROTEOMES OF HEALTHY, POTATO VIRUS A- AND AGROBACTERIUM -INFECTED NICOTIANA BENTHAMIANA PLANTS

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Prunus necrotic ringspot virus

Project description:Prunus necrotic ringspot virus Variation

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