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.
Project description:To extend our understanding of systemic necrosis in susceptible potato tubers infected with the necrotic strain of Potato virus Y (PVYNTN) gene expression was compared between healthy and infected non-necrotic and necrotic (both non-necrotic and necrotic tissue) potato tubers.
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.
Project description:Potato yellow vein virus (PYVV) was detected by RT-PCR in potatoes grown in the Central Colombian highlands, north of Bogotá (~3000 mt height). At this altitude viral whitefly vectors are largely absent, but infection persists because of the use of uncertified tubers. Plants with typical PYVV-induced yellowing symptoms, as well as with atypical yellowing or non-symptomatic were sampled at three separate geographical locations. And five of them were subjected to Next Generation Sequencing (NGS) of their small RNA (sRNA) populations. Contigs to any virus were assembled, and complete or almost complete sequences of four PYVV isolates were thus re-constructed, all from symptomatic plants. Three viral isolates infected plants singly, while the fourth one co-infected the plant together with a potyvirus (potato virus Y, PVY). Relative proportions of sRNAs to each of the three viral genomic RNAs were assessed and found to remain comparable between the four infections. Genomic regions were identified as hotspots to sRNA formation, or as regions that induced poorly sRNAs. Furthermore, PYVV titers in the mixed vs. the single infections were found to be remained comparable, indicating absence of synergistic/antagonistic effect of the potyvirus on the accumulation of PYVV.
Project description:Objectives: Our work focuses on the responses of Solanaceous plants to viruses that cause economically important diseases in tree fruits. Using mock inoculated leaf tissue as a reference, we plan to compare the gene expression profiles of Nicotiana Benthamiana plants infected with one of three viruses; Plum Pox Potyvirus (PPV), Tomato Ringspot Nepovirus (ToRSV), and Prunus Nectrotic Ringspot Nepovirus (PNRSV). Our goals are as follows: (1) Identify genes that are induced/repressed in response to individual viruses. (2) Identify genes that are induced/repressed in response to all 3 viruses. (3) Compare results to existing potato array data to look for similarities in responses to other pathogens. Experimental Design: Nicotiana benthamiana plants were inoculated with one of three viruses: PPV, ToRSV, or PNRSV. 3 week old plants were inoculated by rubbing virus infected plant sap onto leaves dusted with carborundum. Control plants were mock inoculated using sap from healthy plants. All plants were maintained in a growth chamber at 22C for 18 days. 8 plants were inoculated with each virus or mock inoculated. This experiment was repeated twice. 4 biological replicates derived from 2 virus infected plants from each replica experiment (4 plants) are to be used for hybridizations. RNA from all mock inoculated plants was similarly pooled to create 4 biological replicates. Each replicate control will serve as a universal reference sample that is to be hybridized pair wise with each of the three virus infected samples. RNA extraction: After 18 days, un-inoculated leaves displaying clear symptoms were harvested and immediately frozen in liquid N2. Total RNA was purified using Trizol according to TIGRs listed protocol. RNA was subsequently treated with Turbo DNA-free RNase (Ambion cat#1907). Finally, total RNA was further purified on RNeasy columns (Qiagen) according to manufacturer’s instructions and quantified using a Nanodrop spectrophotometer. Keywords: Reference design 23 hybs total
Project description:Nicotiana benthamiana is an important model plant for plant-microbe interaction studies. We compared the proteomes of ribosomes purified from healthy N. benthamiana plants and plants that were infected with two plant pathogens potato virus A (PVA genus Potyvirus) and A. tumefaciens bacteria. Ribosomes were affinity purified from transgenic leaves that expressed FLAG-tagged RPL18B (RPL: ribosome protein large subunit) of A. thaliana. Control purifications were made from non-transgenic plants that were infected with PVA. Our riboproteome revealed ~6600 r-protein hits representing 424 distinct r-proteins that were members from 71 of the expected 81 r-protein families.
Project description:Viral fitness correlates with the magnitude and direction of the perturbation induced in the host's transcriptome: the tobacco etch potyvirus - tobacco case study.
Project description:Objectives: Our work focuses on the responses of Solanaceous plants to viruses that cause economically important diseases in tree fruits. Using mock inoculated leaf tissue as a reference, we plan to compare the gene expression profiles of Nicotiana Benthamiana plants infected with one of three viruses; Plum Pox Potyvirus (PPV), Tomato Ringspot Nepovirus (ToRSV), and Prunus Nectrotic Ringspot Nepovirus (PNRSV). Our goals are as follows: (1) Identify genes that are induced/repressed in response to individual viruses. (2) Identify genes that are induced/repressed in response to all 3 viruses. (3) Compare results to existing potato array data to look for similarities in responses to other pathogens. Experimental Design: Nicotiana benthamiana plants were inoculated with one of three viruses: PPV, ToRSV, or PNRSV. 3 week old plants were inoculated by rubbing virus infected plant sap onto leaves dusted with carborundum. Control plants were mock inoculated using sap from healthy plants. All plants were maintained in a growth chamber at 22C for 18 days. 8 plants were inoculated with each virus or mock inoculated. This experiment was repeated twice. 4 biological replicates derived from 2 virus infected plants from each replica experiment (4 plants) are to be used for hybridizations. RNA from all mock inoculated plants was similarly pooled to create 4 biological replicates. Each replicate control will serve as a universal reference sample that is to be hybridized pair wise with each of the three virus infected samples. RNA extraction: After 18 days, un-inoculated leaves displaying clear symptoms were harvested and immediately frozen in liquid N2. Total RNA was purified using Trizol according to TIGRs listed protocol. RNA was subsequently treated with Turbo DNA-free RNase (Ambion cat#1907). Finally, total RNA was further purified on RNeasy columns (Qiagen) according to manufacturer’s instructions and quantified using a Nanodrop spectrophotometer. Keywords: Reference design
Project description:Potato genotypes from a diploid potato population were divided in two groups based on their response to Potato virus A (PVA). Plants exhibiting hypersensitive response were compared to plants exhibiting non-necrotic response (i.e. blocking virus movement without cell death).<br>The comparisons were made before inoculation and 12 and 24 hours post-inoculation.<br>