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Proteomic analysis of early stage infection in the tobacco-Tobacco mosaic virus (TMV)

ABSTRACT: The early events of virus infection is one of the more poorly understood areas of plant virology and studies on the effect of virus on the host proteome at very early stages of infection are lacking. In the present study, we analysed the proteome on the early stage changes at 15 minutes post inoculation in the tobacco-TMV pathosystem with and without exogenous application of dsRNA p126

INSTRUMENT(S): Q Exactive

ORGANISM(S): Nicotiana Tabacum (common Tobacco)

TISSUE(S): Leaf

SUBMITTER: Sebastien Carpentier

LAB HEAD: sebastien carpentier

PROVIDER: PXD022517 | Pride | 2021-05-12

REPOSITORIES: pride

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			Action	DRS
	20160324_naga_sample_0h_01.raw	Raw
	20160324_naga_sample_0h_02.raw	Raw
	20160324_naga_sample_0h_03.raw	Raw
	20160324_naga_sample_0h_04.raw	Raw
	20160324_naga_sample_0h_05.raw	Raw

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Project description:Here we performed a transcriptomic study on complete symptom development process of CMV-infected Nicotiana tabacum using Solexa/Illumina's high-throughput digital gene expression (DGE) system. 12 DGE libraries (from six virus-infected samples and six corresponding mock-inoculated samples) were constructed, and the gene expression variations between the virus-infected sample and the mock-inoculated sample in each symptom stage were compared. Thousands of differentially expressed genes were obtained by the comparison, and KEGG pathway analysis of these genes suggested that many biological processes (such as phtosynthesis, pigment metabolism and plant-pathogen interaction) were related to the symptom development. The authenticity of the DGE data was further confirmed by analyzing real-time RT-PCR using several random-selected genes. We sequenced a cDNA library constructed from mixture of total RNA from six virus-infected samples and six mock-inoculated samples to get gene information for tobacco leaves in different symptom stages, including vein clearing, mosaic, severe chlorosis, partial recovery, total recovery and re-mosaic, and 95,916 Unigenes were obtained Tobacco leaves were inoculated by CMV-infected leaf homogenate and healthy leaf homogenate, respectivley. Six time points with different symptom stage were selected, and one virus-infect sample and one mock-inoculated sample were collected at each time. In order to average out variation of different plants, five leaves from five different plants were mixed to prepare every RNA sample. Twelve individual tag libraries of samples (six infected samples and six mock-inoculated samples) were constructed in parallel. For the gene expression analysis, the twelve samples were grouped into six groups, and each group contained a virus-infected sample and a mock-inoculated sample collected at the same time. In each group, the DGE data of virus-infected sample were compared to that of mock-inoculated sample to obtain the gene expression variations. Illumina sequencing of transcripts from virus-infected and mock-inoculated samples to get gene information for tobacco leaves in different symptom stages. In order to get more gene information, the systemically infected leaves and mock-inoculated leaves were harvested at six time points and five leaves from five different plants were collected at each time point. The RNA-Seq analysis provided gene information for mock-inoculated and virus-infected tobacco leaves in different symptom stages.

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Proteomic analysis of early stage infection in the tobacco-Tobacco mosaic virus (TMV)

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