Project description:Purpose and strategy: Grapevine fanleaf virus (GFLV) causes variable symptoms in most vineyards worldwide. To better understand GFLV-grapevine interactions in relation to symptom development, field and greenhouse trials were conducted with a grapevine genotype that exhibits distinct symptoms in response to a severe and a mild strain of GFLV. Results: After validation of the infection status of the experimental vines by high throughput sequencing, the transcriptomic and metabolomic profiles in plants infected with the two viral strains were tested and compared by RNA-Seq and LC-MS, respectively, in the differentiating grapevine genotype. In vines infected with the severe GFLV strain, 1,023 genes, among which some are implicated in the regulation of the hypersensitive-type response, were specifically de-regulated, and a higher accumulation of resveratrol and phytohormones was observed. Interestingly, some experimental vines restricted the virus to the rootstock and remained symptom-less. Our results suggest that GFLV induces a strain- and cultivar-specific defense reaction similar to a hypersensitive reaction. This type of defense leads to a severe stunting phenotype in some grapevines whereas others are resistant. This work is the first evidence of a hypersensitive-like reaction in grapevine during virus infection. Conclusion: Our results suggest that GFLV induces a strain- and cultivar-specific defense reaction similar to a hypersensitive reaction. This type of defense leads to a severe stunting phenotype in some grapevines whereas others are resistant. This work is the first evidence of a hypersensitive-like reaction in grapevine during virus infection.
Project description:Nicotiana benthamiana was infected with several strains of grapevine fanleaf virus (GFLV). Apical tissue was collected 4, 7, and 12 days after inoculation, with identical samples for shotgun proteomics and transcriptomics analysis. Five leaf discs per leaf were collected a pooled by three plants into a single tube at each time point. Five biological replicates represent each treatment at each time point for a total of 75 samples. Two samples were lost between sample processing and data acquisition. The analysis methods between proteomics and transcriptomics were then cross-analyzed for host genes responsible for phenotypic differences upon infection.
Project description:Transcriptome sequencing from Nicotiana benthamiana leaves non-infected and infected with Turnip mosaic virus at 6 days post inoculation.
Project description:Micrarray analysis was used to identify gene expression changes associated with disease development and virus movement in N.benthamina plants induced by infection with the SACMV 1-Plex , 385K array Nicotiana benthamiana (NimbleGen design name: 110121_N_benthamiana_60mer_exp) was used in this study to monitor changes in gene expression levels in SACMV- infected leaf tissue. Three biological replicates were used for infected leaf tissue and one pooled mock-inoculated sample was used as a control/reference.
Project description:Transcriptomes of wild-type Nicotiana benthamiana plants inoculated with plum pox virus (PPV) or the P1Pro clone, a PPV deletion mutant that lacks the self-cleavage inhibitory domain of the P1 leader protease; in addition, N. benthamiana nahG-expressing plants inoculated with P1Pro were analyzed to identify genes whose expression is altered by P1Pro infections but does not depend on salicylic acid signaling.
Project description:Hypersensitive response-related programmed cell death (PCD) has been extensively analyzed in various plant–virus interactions. However, little is known about changes in gene expression associated with cell death caused by compatible viruses. The synergistic interaction of Potato virus X (PVX) with Plum pox virus (PPV) results in increased symptoms that lead to systemic necrosis (SN) in Nicotiana benthamiana. Here, we performed three transcriptome comparisons in response to i) a PVX recombinant virus expressing the helper component-proteinase (HC-Pro) gene from PPV that leads to SN, ii) a systemic incompatible interaction conferred by the Tobacco mosaic virus (TMV)-resistance gene N (SHR), and iii) the depletion of the PBE subunit of the proteasome that leads to PCD by virus induced gene silencing (VIGS Prot), at early and late stages of infection. Our analysis indicated that the SN response was clustered with SHR by the similarity of their overall gene expression profiles. However, the expression profiles of defence-related and hormone-responsive genes in response to SN were more closely related to the response to VIGS Prot than to that elicited by SHR. This suggests the potential contribution of proteasome dysfunction to the increase in pathogenicity observed in PVX-potyvirus infections We compare the gene expression profiles of Nicotiana benthamiana plants infected with either necrosis-inducing viruses or non-necrosis-inducing viruses, as follow: PVX/HCWT-infected plants versus plants infected with a PVX recombinant virus expressing a PPV HC-Pro mutant (PVX/HCLH) that was unable to induce the SN response (SN comparison), at 7 and 11 days postinoculation (dpi), (ii) TRV:NbPBE-silenced plants versus plants infected with the TRV empty vector (VIGS Prot comparison), at 4 and 8 days after infiltration (dpa), and (iii) TMV-GFP-infected, N-transgenic plants versus wild-type plants infected with TMV-GFP (SHR comparison), at 24 and 72 hours after temperature shift (hts). Per time and treatment, three independent biological replicates were used to monitor differences in gene expression between treatments
