Our goal was to target silencing of the Plum pox virus coat protein (PPV CP) gene independently expressed in plants. Clone C-2 is a transgenic plum expressing CP. We introduced and verified, in planta, the effects of the inverse repeat of CP sequence split by a hairpin (IRSH) that was characterized in the HoneySweet plum. The IRSH construct was driven by two CaMV35S promoter sequences flanking the CP sequence and had been introduced into C1738 plum. To determine if this structure was enough to induce silencing, cross-hybridization was made with the C1738 clone and the CP expressing but PPV-susceptible C2 clone. In total, 4 out of 63 clones were silenced. While introduction of the IRSH is reduced due to the heterozygous character in C1738 plum, the silencing induced by the IRSH PPV CP is robust. Extensive studies, in greenhouse containment, demonstrated that the genetic resource of C1738 clone can silence the CP production. In addition, these were verified through the virus transgene pyramiding in the BO70146 BlueByrd cv. plum that successfully produced resistant BlueByrd BO70146 × C1738 (HybC1738) hybrid plums.
- MeSH
- Biotechnology methods MeSH
- Genetic Engineering methods MeSH
- Disease Resistance * MeSH
- Prunus genetics virology MeSH
- Transgenes MeSH
- Gene Silencing * MeSH
- Capsid Proteins genetics metabolism MeSH
- Plum Pox Virus genetics pathogenicity MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Plants of the genus Pleione, originating from hobby growers in the Netherlands and in the Czech Republic, were conspicuous for viral infection, showing symptoms of leaf mosaic or flower breaking. Using Sanger and high throughput sequencing, the full genome sequence of a novel potyvirus was obtained from sequencing data. The genome sequence was annotated and compared to the genome of other potyviruses. The virus was experimentally transmitted by aphids into Pleione and Chenopodium quinoa plants. The name Pleione flower breaking virus (PlFBV) was suggested for the new virus. The presence of the virus was confirmed using RT-PCR, with newly designed primers targeting this new species. The incidence of the virus was contrasted between both countries and might have been influenced by the growth conditions and the exposure of the plants to aphids.
- MeSH
- Molecular Sequence Annotation MeSH
- Chenopodium quinoa virology MeSH
- Insect Vectors MeSH
- Incidence MeSH
- Aphids MeSH
- Plant Diseases virology MeSH
- Orchidaceae virology MeSH
- Reverse Transcriptase Polymerase Chain Reaction MeSH
- Potyvirus classification genetics isolation & purification MeSH
- Disease Transmission, Infectious MeSH
- Sequence Analysis, DNA MeSH
- Whole Genome Sequencing MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
- Netherlands MeSH
Potato virus Y (PVY) is among the most economically important plant pathogens. Using cryoelectron microscopy, we determined the near-atomic structure of PVY's flexuous virions, revealing a previously unknown lumenal interplay between extended carboxyl-terminal regions of the coat protein units and viral RNA. RNA-coat protein interactions are crucial for the helical configuration and stability of the virion, as revealed by the unique near-atomic structure of RNA-free virus-like particles. The structures offer the first evidence for plasticity of the coat protein's amino- and carboxyl-terminal regions. Together with mutational analysis and in planta experiments, we show their crucial role in PVY infectivity and explain the ability of the coat protein to perform multiple biological tasks. Moreover, the high modularity of PVY virus-like particles suggests their potential as a new molecular scaffold for nanobiotechnological applications.
- MeSH
- Capsid chemistry metabolism MeSH
- Protein Conformation * MeSH
- Models, Molecular * MeSH
- Plant Diseases virology MeSH
- Potyvirus physiology ultrastructure MeSH
- RNA, Viral chemistry metabolism MeSH
- Amino Acid Sequence MeSH
- Protein Binding MeSH
- Binding Sites MeSH
- Virion MeSH
- Capsid Proteins chemistry metabolism MeSH
- Structure-Activity Relationship MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Plum pox virus (PPV, family Potyviridae) is one of the most important viral pathogens of Prunus spp. causing considerable damage to stone-fruit industry worldwide. Among the PPV strains identified so far, only PPV-C, PPV-CR, and PPV-CV are able to infect cherries under natural conditions. Herein, we evaluated the pathogenic potential of two viral isolates in herbaceous host Nicotiana benthamiana. Significantly higher accumulation of PPV capsid protein in tobacco leaves infected with PPV-CR (RU-30sc isolate) was detected in contrast to PPV-C (BY-101 isolate). This result correlated well with the symptoms observed in the infected plants. To further explore the host response upon viral infection at the molecular level, a comprehensive proteomic profiling was performed. Using reverse-phase ultra-high-performance liquid chromatography followed by label-free mass spectrometry quantification, we identified 38 unique plant proteins as significantly altered due to the infection. Notably, the abundances of photosynthesis-related proteins, mainly from the Calvin-Benson cycle, were found more aggressively affected in plants infected with PPV-CR isolate than those of PPV-C. This observation was accompanied by a significant reduction in the amount of photosynthetic pigments extracted from the leaves of PPV-CR infected plants. Shifts in the abundance of proteins that are involved in stimulation of photosynthetic capacity, modification of amino acid, and carbohydrate metabolism may affect plant growth and initiate energy formation via gluconeogenesis in PPV infected N. benthamiana. Furthermore, we suggest that the higher accumulation of H2O2 in PPV-CR infected leaves plays a crucial role in plant defense and development by activating the glutathione synthesis.
- MeSH
- Chlorophyll biosynthesis MeSH
- Chromatography, Reverse-Phase MeSH
- Energy Metabolism genetics MeSH
- Photosynthesis genetics MeSH
- Genotype MeSH
- Glutathione biosynthesis MeSH
- Mass Spectrometry MeSH
- Host-Pathogen Interactions genetics MeSH
- Carotenoids biosynthesis MeSH
- Plant Leaves genetics metabolism virology MeSH
- Plant Diseases genetics virology MeSH
- Oxidation-Reduction MeSH
- Hydrogen Peroxide metabolism MeSH
- Heat-Shock Proteins classification genetics metabolism MeSH
- Prunus avium virology MeSH
- Gene Expression Regulation, Plant * MeSH
- Plant Proteins classification genetics metabolism MeSH
- Prunus domestica virology MeSH
- Nicotiana genetics metabolism virology MeSH
- Plum Pox Virus classification genetics growth & development pathogenicity MeSH
- Chromatography, High Pressure Liquid MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Pea seed-borne mosaic virus (PSbMV) significantly reduces yields in a broad spectra of legumes. The eukaryotic translation initiation factor has been shown to confer resistance to this pathogen, thus implying that translation and proteome dynamics play a role in resistance. This study presents the results of a proteome-wide analysis of Pisum sativum L. response to PSbMV infection. LC-MS profiling of two contrasting pea cultivars, resistant (B99) and susceptible (Raman) to PSbMV infection, detected >2300 proteins, 116 of which responded to PSbMV ten and/or twenty days post-inoculation. These differentially abundant proteins are involved in number of processes that have previously been reported in the plant-pathogen response, including protein and amino acid metabolism, stress signaling, redox homeostasis, carbohydrate metabolism, and lipid metabolism. We complemented our proteome-wide analysis work with targeted analyses of free amino acids and selected small molecules, fatty acid profiling, and enzyme activity assays. Data from these additional experiments support our findings and validate the biological relevance of the observed proteome changes. We found surprising similarities in the resistant and susceptible cultivars, which implies that a seemingly unaffected plant, with no detectable levels of PSbMV, actively suppresses viral replication. BIOLOGICAL SIGNIFICANCE: Plant resistance to PSbMV is connected to translation initiation factors, yet the processes involved are still poorly understood at the proteome level. To the best of our knowledge, this is the first survey of the global proteomic response to PSbMV in plants. The combination of label-free LC-MS profiling and two contrasting cultivars (resistant and susceptible) provided highly sensitive snapshots of protein abundance in response to PSbMV infection. PSbMV is a member of the largest family of plant viruses and our results are in accordance with previously characterized potyvirus-responsive proteomes. Hence, the results of this study can further extend our knowledge about these pathogens. We also show that even though no viral replication is detected in the PSbMV-resistant cultivar B99, it is still significantly affected by PSbMV inoculation.
- MeSH
- Chromatography, Liquid MeSH
- Eukaryotic Initiation Factors MeSH
- Pisum sativum virology MeSH
- Host-Pathogen Interactions * MeSH
- Plant Diseases virology MeSH
- Disease Resistance genetics MeSH
- Potyvirus pathogenicity MeSH
- Proteomics methods MeSH
- Plant Viruses MeSH
- Tandem Mass Spectrometry MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Plum pox virus (PPV), the agent responsible for sharka disease, is the most important viral pathogen of stone fruit trees. The fruits of these fruit species are widely used in the processing industry, thus being economically very attractive. This viral disease significantly reduces the vitality of the fruit trees and the quantity and quality of fruits. The present review describes recent methods used for the identification and characterization of economically important Plum pox virus. Understanding the diversity of plant viruses is an essential step to design efficient management strategies to eliminate economical losses.
BACKGROUND: The eukaryotic translation initiation factor 4E was shown to be involved in resistance against several potyviruses in plants, including pea. We combined our knowledge of pea germplasm diversity with that of the eIF4E gene to identify novel genetic diversity. METHODOLOGY/PRINCIPAL FINDINGS: Germplasm of 2803 pea accessions was screened for eIF4E intron 3 length polymorphism, resulting in the detection of four eIF4E(A-B-C-S) variants, whose distribution was geographically structured. The eIF4E(A) variant conferring resistance to the P1 PSbMV pathotype was found in 53 accessions (1.9%), of which 15 were landraces from India, Afghanistan, Nepal, and 7 were from Ethiopia. A newly discovered variant, eIF4E(B), was present in 328 accessions (11.7%) from Ethiopia (29%), Afghanistan (23%), India (20%), Israel (25%) and China (39%). The eIF4E(C) variant was detected in 91 accessions (3.2% of total) from India (20%), Afghanistan (33%), the Iberian Peninsula (22%) and the Balkans (9.3%). The eIF4E(S) variant for susceptibility predominated as the wild type. Sequencing of 73 samples, identified 34 alleles at the whole gene, 26 at cDNA and 19 protein variants, respectively. Fifteen alleles were virologically tested and 9 alleles (eIF4E(A-1-2-3-4-5-6-7), eIF4E(B-1), eIF4E(C-2)) conferred resistance to the P1 PSbMV pathotype. CONCLUSIONS/SIGNIFICANCE: This work identified novel eIF4E alleles within geographically structured pea germplasm and indicated their independent evolution from the susceptible eIF4E(S1) allele. Despite high variation present in wild Pisum accessions, none of them possessed resistance alleles, supporting a hypothesis of distinct mode of evolution of resistance in wild as opposed to crop species. The Highlands of Central Asia, the northern regions of the Indian subcontinent, Eastern Africa and China were identified as important centers of pea diversity that correspond with the diversity of the pathogen. The series of alleles identified in this study provides the basis to study the co-evolution of potyviruses and the pea host.
- MeSH
- Alleles MeSH
- Eukaryotic Initiation Factor-4E genetics physiology MeSH
- Pisum sativum genetics metabolism virology MeSH
- Plant Diseases virology MeSH
- Potyvirus pathogenicity MeSH
- Plant Proteins genetics physiology MeSH
- Geography MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
Members the Potyviridae family belong to a group of plant viruses that are causing devastating plant diseases with a significant impact on agronomy and economics. Plum pox virus (PPV), as a causative agent of sharka disease, is widely discussed. The understanding of the molecular biology of potyviruses including PPV and the function of individual proteins as products of genome expression are quite necessary for the proposal the new antiviral strategies. This review brings to view the members of Potyviridae family with respect to plum pox virus. The genome of potyviruses is discussed with respect to protein products of its expression and their function. Plum pox virus distribution, genome organization, transmission and biochemical changes in infected plants are introduced. In addition, techniques used in PPV detection are accentuated and discussed, especially with respect to new modern techniques of nucleic acids isolation, based on the nanotechnological approach. Finally, perspectives on the future of possibilities for nanotechnology application in PPV determination/identification are outlined.
- MeSH
- Genome, Viral MeSH
- Plant Diseases virology MeSH
- Plum Pox Virus physiology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
The complete nucleotide sequence of the ssRNA genome of a lupine potyvirus (LP) isolate was determined. It comprised 10,113 nucleotides excluding the poly(A) tail. Phylogenetic analysis of CP protein sequences identified pepper veinal mottle virus, narcissus yellow stripe virus, and chili veinal mottle virus as the closest relatives, sharing coat protein amino acid sequence identities of only about 64% with the LP isolate. Thus, LP can be regarded as a member of a newly described potyvirus species, for which the name Lupine mosaic virus (LuMV) is proposed.
- MeSH
- Phylogeny MeSH
- Genome, Viral MeSH
- Lupinus virology MeSH
- Plant Diseases virology MeSH
- Potyvirus genetics isolation & purification MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
Eight isolates of the Pea seed-borne mosaic virus (PSbMV) from the Czech Republic were studied regarding their biological and molecular characteristics. Molecular characterization using RT-PCR was done on the 5'(Nter)NIb-CP-UTR3' region amplified using universal CPUP/P9502 primer pair and the newly designed PSB8812/PSB944, and PSB8800/PSB9440 primer pairs, respectively. Sequential and phylogenetic analysis of CP-UTR3' region from all isolates showed that the available Czech and GenBank PSbMV isolates were distributed into 4 clusters in agreement with their diversification and according to their biological characteristics (i.e. pathotype). The molecular data were confirmed by biological testing on different pea cultivars. The Czech isolates were distributed into two pathotypes, the P-1 (7 isolates) and P-4 (1 isolate).
- MeSH
- Phylogeny MeSH
- Genetic Variation MeSH
- Pisum sativum virology MeSH
- Molecular Sequence Data MeSH
- Plant Diseases virology MeSH
- Potyvirus chemistry classification genetics isolation & purification MeSH
- Amino Acid Sequence MeSH
- Sequence Alignment MeSH
- Viral Proteins chemistry genetics MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
