Comparison of resistance to potyviruses within Solanaceae: infection of potatoes with tobacco etch potyvirus and peppers with potato A and Y potyviruses

The reaction of several cultivated potato varieties (Solarium tuberosum L.) to three strains of tobacco etch potyvirus (TEV-F, TEV-Mex21 and TEV-ATCC) and the reaction of several pepper lines (Capsicum annuum L. and C. chinense L.) to two strains of potato Y potyvirus (PVY^O and PVY^N) and one strain of potato A potyvirus (PVA-M) was tested. The potato varieties included in this study carried resistance genes against PVY, PVA and potato V potyvirus, but all were susceptible to TEV and developed mottle and mosaic symptoms. TEV was readily transmitted by mechanical inoculation from tobacco and potato to potato, whereas transmission from pepper to potato occurred infrequently. TEV was transmitted through potato tubers, and from pepper to potato plants by aphids. Lack of detectable systemic infection following graft-inoculation indicated extreme resistance to PVY^O and PVA in several pepper lines. No pepper line was systemically infected with PVY^N following mechanical inoculation (graft-inoculation was not carried out with PVY^N). The development of necrotic lesions following mechanical and graft-inoculation indicated hypersensitive response to PVY^O in several pepper lines which resembled the resistance responses to these potyvirus strains in potato. Results of this study together with previous work indicate that C. annuum cv. Avelar is resistant to four potyviruses [PVY, PVA, pepper mottle potyvirus (PepMoV) and some isolates of TEV]; C. annuum cv. Criollo de Morelos and C. chinense PI 152225 and PI 159236 are resistant to three potyviruses (PVY, PepMoV and PVA; and PVY, PepMoV and TEV, respectively); C. annuum 9093–1 and 92016–1 are resistant to PVY and PepMoV; and C. annuum cv. Jupiter and C. annuum cv. RNaky are resistant to PVY^N and PVA.     

Genetic variability of Potato virus Y (PVY) and Tobacco etch virus (TEV) from naturally infected pepper fields in the Hatay region of Turkey

The genetic structure of Potato virus Y (PVY) and Tobacco etch virus (TEV) (Potyvirus) populations was investigated in pepper fields in two regions in Turkey. The diversity of PVY and TEV populations according to coat protein (CP) and VPg coding regions showed some similarity. All the isolates built a monophyletic group due to a single introduction event or multiple introductions of genetically similar isolates. All the isolates of both viruses showed evidence to the diversification for a long time. Based on VPg and CP sequences, all PVY isolates corresponded to clade C1. Turkish potyvirus isolates were only able to break the pvr2¹ resistance allele and therefore belonged to pathotype (0,?1). The Pvr4 dominant gene was found to be efficient and durable against PVY but not at all efficient against TEV. Consequently, the pvr2² resistance allele, efficient resistance against PVY and TEV pathotype (0,?1) isolates, would be the most suitable strategy to control potyviruses.     

Trichome density of main stem is tightly linked to PepMoV resistance in chili pepper (Capsicum annuum L.)

A relationship between pepper trichome and pepper mottle virus (PepMoV) resistance was examined. In an intraspecific F(2) mapping population from the cross between Capsicum annuum CM334 (trichome-bearing and PepMoV resistant) and Chilsungcho (glabrous and PepMoV susceptible), major QTLs for both traits were identified by composite interval mapping in linkage group (LG) 24 corresponding a telomere region on pepper chromosome 10. Ptel1 of putative trichome enhancing locus was a common major QTL for trichome density on the main stem and calyx. Ptel1 apart from HpmsE031 at a 1.03?cM interval was specifically associated to the trichome density on the main stem, whereas Ptel2 near m104 marker on LG2 was specific for the calyx trichome. Epistatic analysis indicated that Ptel1 engaged in controlling the trichome density by mutual interactions with the organ-specific QTLs. For PepMoV resistance, two QTLs (Pep1 and Pep2) were identified on the LG 24. Pep1 was located with Ptel1 in the R-gene cluster (RGC) for potyvirus resistance including Pvr4 with broad spectrum resistance to potyviruses. Pep1 flanking TG420 marker seemed to be the major factors determining correlation with PepMoV resistance. These results indicate that the level of trichome density on pepper main stem can be used as a morphological marker for Pvr4 in pepper breeding.     

Identification and comparative mapping of a dominant potyvirus resistance gene cluster in Capsicum

The dominant gene Pvr7 from Capsicum chinense Jacq. ’PI159236’ confers resistance to the pepper mottle potyvirus (PepMoV) Florida (V1182) strain. This gene is tightly linked to the dominant potyvirus resistance gene Pvr4 with observed recombination frequencies of 0.012 to 0.016. A cleaved amplified polymorphic sequence (CAPS) marker linked to Pvr4 was used to localize Pvr4 and, by extension, Pvr7, to linkage group 10 on an interspecific map of pepper. Our results indicated that Pvr4, Pvr7, and Tsw, a gene conferring resistance to tomato spotted wilt virus, comprise the first identified cluster of dominant disease resistance genes in Capsicum L. This position does not correspond to the locations of dominant potyvirus resistance genes in potato or to the positions of any other mapped solanaceous resistance genes or resistance gene homologues. Received: 20 September 1999 / Accepted: 21 March 2000     

Development of a CAPS marker for the Pvr4 locus: a tool for pyramiding potyvirus resistance genes in pepper.

The Pvr4 resistance gene in pepper confers a complete resistance to the three pathotypes of potato virus Y (PVY) and to pepper mottle virus (PepMoV). In order to use this gene in a marker-assisted selection (MAS) program and to permit the pyramiding of several potyvirus resistance genes in the same cultivar, tightly linked amplified fragment length polymorphism (AFLP) markers were obtained by the bulked segregant analysis method. Eight linked AFLP markers were mapped in an interval from 2.1 +/- 0.8 to 13.8 +/- 2.9 cM around this locus. The closest codominant AFLP marker was converted into a codominant CAPS (cleaved amplified polymorphic sequence) marker using data from the alignment of the two allele sequences. We have further characterized the relevance of the CAPS marker for MAS programs in different pepper breeding lines.     

A new chrysanthemum potyvirus: molecular evidence

Abstract

A number of viruses are known to infect chrysanthemum plants, however in the present study a previously unknown potyvirus was detected using techniques such as ELISA, RT-PCR and hybridization. The ELISA-positive samples were amplified using a potyvirus group-specific primer which gave an amplification of ～850 bp. The amplified product was cloned and sequenced, and shows 72 – 73% homology with known potyviruses that infect chrysanthemums such as Potato virus Y potyvirus, Soyabean mosaic virus and Turnip mosaic potyvirus when compared to the sequence available in the database. However, present potyvirus isolates show 93% homology with Chilli veinal mottle virus and Pepper vein banding virus. The results were further confirmed by Northern hybridization. This is the first report of a potyvirus similar to Chilli veinal mottle virus, and Pepper vein banding virus infecting chrysanthemums.     

Development of RAPD and SCAR markers linked to the Pvr4 locus for resistance to PVY in pepper ( Capsicum annuum L.)

Potato Virus Y (PVY) is the only potyvirus infecting pepper ( Capsicum annuum L.) in Europe. Currently, the development of pepper varieties resistant to PVY seems to be the most-efficient method to control PVY damage. Among the sources of resistance, a monogenic dominant gene Pvr4 confers resistance against all known PVY pathotypes. In this work, bulked segregant analysis (BSA) was used to search for randomly amplified polymorphic DNA (RAPD) markers linked to the Pvr4 gene, using segregating progenies obtained by crossing a homozygous resistant ('Serrano Criollo de Morelos-334') with a homozygous susceptible ('Yolo Wonder') cultivar. Eight hundred decamer primers were screened to identify one RAPD marker (UBC19(1432)) linked in repulsion phase to Pvr4. This marker was converted into a dominant sequence characterised amplified region (SCAR) marker (SCUBC19(1423)). This marker was mapped into a dense Capsicum genetic map in a region where several genes for resistance to different diseases are located. This marker can be useful to identify PVY-resistant genotypes in segregating progenies of pepper in marker-assisted selection (MAS) breeding programs.     

Relocation of the NIb Gene in the Tobacco Etch Potyvirus Genome

Potyviruses express most of their proteins from a long open reading frame that is translated into a large polyprotein processed by three viral proteases. To understand the constraints on potyvirus genome organization, we relocated the viral RNA-dependent RNA polymerase (NIb) cistron to all possible intercistronic positions of the Tobacco etch virus (TEV) polyprotein. Only viruses with NIb at the amino terminus of the polyprotein or in between P1 and HC-Pro were viable in tobacco plants.     

Soybean RNA interference lines silenced for eIF4E show broad potyvirus resistance

Soybean mosaic virus (SMV), a potyvirus, is the most prevalent and destructive viral pathogen in soybean-planting regions of China. Moreover, other potyviruses, including bean common mosaic virus (BCMV) and watermelon mosaic virus (WMV), also threaten soybean farming. The eukaryotic translation initiation factor 4E (eIF4E) plays a critical role in controlling resistance/susceptibility to potyviruses in plants. In the present study, much higher SMV-induced eIF4E1 expression levels were detected in a susceptible soybean cultivar when compared with a resistant cultivar, suggesting the involvement of eIF4E1 in the response to SMV by the susceptible cultivar. Yeast two-hybrid and bimolecular fluorescence complementation assays showed that soybean eIF4E1 interacted with SMV VPg in the nucleus and with SMV NIa-Pro/NIb in the cytoplasm, revealing the involvement of VPg, NIa-Pro, and NIb in SMV infection and multiplication. Furthermore, transgenic soybeans silenced for eIF4E were produced using an RNA interference approach. Through monitoring for viral symptoms and viral titers, robust and broad-spectrum resistance was confirmed against five SMV strains (SC3/7/15/18 and SMV-R), BCMV, and WMV in the transgenic plants. Our findings represent fresh insights for investigating the mechanism underlying eIF4E-mediated resistance in soybean and also suggest an effective alternative for breeding soybean with broad-spectrum viral resistance.     

An Ry-mediated resistance response in potato requires the intact active site of the NIa proteinase from potato virus Y

Ry confers extreme resistance to all strains of potato virus Y (PVY). To identify the elicitor of the Ry-mediated resistance against PVY in potato, we expressed each of the PVY-encoded proteins in leaves of PVY-resistant (Ry) and -susceptible (ry) plants. For most of the proteins tested, there was no evident response. However, when the NIa proteinase was expressed in leaves of Ry plants, there was a hypersensitive response (HR). Proteinase active site mutants failed to induce the Ry-mediated response. The HR was also induced by the NIa proteinase from pepper mottle virus (PepMoV), which has the same cleavage specificity as the PVY enzyme, but not by the tobacco etch virus (TEV) or the potato virus A (PVA) proteinases that cleave different peptide motifs. Based on these results, we propose that Ry-mediated resistance requires the intact active site of the NIa proteinase. Although the structure of the active proteinase could have elicitor activity, it is possible that this proteinase releases an elicitor by cleavage of a host-encoded protein. Alternatively, the proteinase could inactivate a negative regulator of the Ry-mediated resistance response.     

A survey of mixed Begomovirus infection in solanaceae and fabaceae at different altitudes in East Java,Indonesia

A multiplex primer set was developed to detect four Begomoviruses in East Java, Indonesia, i.e. Tomato leaf curl New Delhi virus (ToLCNDV), Tomato yellow leaf curl Kanchanaburi virus (TYLCKaV), Pepper yellow leaf curl Indonesia virus (PepYLCIV) and Mungbean yellow mosaic India virus (MYMIV). Survey at different altitudes found that begomoviruses infecting pepper, tomato and long bean were more variable, while in eggplant and string bean were more uniform. As a single virus, TYLCKaV infected eggplant, and sometimes tomato and pepper; PepYLCIV infected pepper, tomato and long bean; ToLCNDV only infected long bean and tomato at low frequency; and MYMIV infected beans. Mixed infection occurred more frequently in the low altitude areas. Subsequent examination indicated that Cucumber mosaic virus (CMV) and potyviruses were also responsible for diseased fabaceous. Our data suggest a relationship between altitudes and virus species occurrence. However, which viral species infects a crop is mainly influenced by the crop rather than by altitude.     

Die Bedeutung elektrophoretischer Proteinanalysen bei phytopathogenen Pilzen als Hilfsmittel zu ihrer taxonomischen Charakterisierung

Sunflower is one of the most important oilseed crops, which is grown in many countries. Increasing demand to use sunflower oil has expanded the under cultivation area throughout the world. Sunflower viruses have been reported as one of the pathogens that reduce the quantity and the quality of this product in a number of countries. In our research to study the occurrence and distribution of viruses in sunflower fields of Iran, 562 samples were collected from different fields in Kerman and Isfahan provinces during growing seasons from 2009 to 2011. Potato virus Y (PVY), Cucumber mosaic virus (CMV) and Tomato spotted wilt virus (TSWV) were detected by double antibody sandwich enzyme-linked immunosorbent assay. Also, numbers of samples were positive for infection to potyviruses (except PVY) in antigen-coated-plate enzyme-linked immunosorbent assay test using potyviruses general antiserum. Infected plants had symptoms like mosaic, yellowing, deformation, necrotic and chlorotic lesions and mottling on leaves and stunting. The infection rates of potyviruses, PVY, CMV and TSWV in Isfahan province were 33, 22, 4.18 and 3.25% of collected samples, respectively. The corresponding rates for samples from Kerman province were 15, 5, 0.8 and 0.4% of collected samples. According to these results, viral infection in Isfahan province was more than surveyed in Kerman province during the mentioned period of three years. Furthermore, generally there was a decrease in the percentage of viral infection during this three growing seasons in both provinces. This is the first report of the detection of CMV and TSWV in sunflower fields of Isfahan and Kerman and first report of PVY in Isfahan.     

Novel resistances to four potyviruses in tuber-bearing potato species, and temperature-sensitive expression of hypersensitive resistance to potato virus Y

Novel potyvirus resistance specificities were found in eight tested wild potato species (clones): hypersensitive resistance (HR) to potato Y potyvirus (PVY) strain groups PVY^O in Solanum megistacrolobum and S. polyadenium and PVY^N in S. stoloniferum; HR to potato V potyvirus (PW) in S. maglia, S. polyadenium, S. stoloniferum, S. sparsipilum and S. sucrense, HR to potato A potyvirus (PVA) strain group 1 in S. sucrense, and extreme resistance (ER) to PVA in S. polyadenium. S. commersonii and S. stoloniferum expressed HR to tobacco etch potyvirus (TEV) which has not been reported previously in potato species. The studied clone of S. stoloniferum expressed HR to all potyviruses and potyvirus strains tested. The clone of S. stoloniferum (2n = 48; nuclear DNA content (2C) = 3.6 pg) and S. chacoense (2n = 24; 2C=1.9 pg) were crossed and one hybrid (2n = 36; 2C = 2.9 pg) was obtained. The hybrid expressed HR to all tested potyviruses except PVA, which indicated that HR to PVA was controlled by a gene which is different from the genes (or gene) controlling HR to PVY^O, PVY^N, PVV and TEV in S. stoloniferum. On the other hand, S. chacoense and the hybrid expressed ER to cucumber mosaic cucumovirus (CMV), whereas S. stoloniferum was susceptible to CMV. All tested wild species and the six tested potato cultivars (S. tuberosum subsp. tuberosum) expressed HR to PVV. Expression of HR following infection with PVY^N induced systemic acquired resistance (SAR) in S. chacoense. HR to PVY^N in S. sparsipilum and S. sucrense and to PVY^O in potato cv. Pito was efficiently expressed at lower temperatures (16/18°C) indicated by the development of distinct necrotic lesions and/or vein necrosis in inoculated leaves, whereas the HR was rendered less effective at higher temperatures (19/24°C) which was indicated by the development of systemic infection with leaf-drop and mosaic symptoms.     

The recessive potyvirus resistance gene <Emphasis Type="Italic">pot-1</Emphasis> is the tomato orthologue of the pepper <Emphasis Type="Italic">pvr2-eIF4E</Emphasis> gene

The translation initiation factor 4E (eIF4E) has been implicated in naturally occurring resistance to Potato virus Y (PVY) determined by the pvr2 locus in pepper (Capsicum annuum). Here, the molecular basis of the recessive resistance to PVY and Tobacco etch virus (TEV) controlled by the pot-1 locus in tomato (Lycopersicon esculentum; now Solanum lycopersicum) was investigated. On the basis of genetic mapping data that indicated that pot-1 and pvr2 are located in syntenic regions of the tomato and pepper genomes, the possible involvement of eIF4E in pot-1-mediated resistance was assessed. Genetic mapping of members of the eIF4E multigenic family in tomato introgression lines revealed that an eIF4E locus indeed maps in the same genomic region as pot-1. By comparing eIF4E coding sequences between resistant and susceptible Lycopersicon genotypes, a small number of polymorphisms that co-segregate with the pot-1 locus were identified, suggesting that this gene could be involved in resistance to potyviruses. Functional complementation experiments using Potato virus X-mediated transient expression of eIF4E from a susceptible genotype in a resistant pepper genotype confirmed that a small number of amino acid substitutions in the eIF4E protein indeed account for resistance/susceptibility to both the PVY and TEV, and consequently that pot-1 and pvr2 are orthologues. Taken together, these results support the role of this eIF4E gene as a key component of recessive resistance to potyviruses, and validate the comparative genomic approach for the molecular characterization of recessive resistance genes.     

Comparative studies of compatible and incompatible pepper–<Emphasis Type="Italic">Tobamovirus</Emphasis> interactions and the evaluation of effects of 24-epibrassinolide

The aim of study was to gain a deeper knowledge about local and systemic changes in photosynthetic processes and sugar production of pepper infected by Obuda pepper virus (ObPV) and Pepper mild mottle virus (PMMoV). PSII efficiency, reflectance, and gas exchange were measured 48 and/or 72 h after inoculation (hpi). Sugar accumulation was checked 72 hpi and 20 d after inoculation (as a systemic response). Inoculation of leaves with ObPV led to appearance of hypersensitive necrotic lesions (incompatible interaction), while PMMoV caused no visible symptoms (compatible interaction). ObPV (but not PMMoV) lowered F_v/F_m (from 0.827 to 0.148 at 72 hpi). Net photosynthesis decreased in ObPV-infected leaves. In ObPV-inoculated leaves, the accumulation of glucose, fructose, and glucose-6-phosphate was accompanied with lowered sucrose, maltoheptose, nystose, and trehalose contents. PMMoV inoculation increased the contents of glucose, maltose, and raffinose in the inoculated leaves, while glucose-6-phosphate accummulated in upper leaves.     

Both common and specific genetic factors are involved in polygenic resistance of pepper to several potyviruses

Absolute resistance to potato virus Y pathotype 0 (PVY 0), potyvirus E and chili veinal mottle virus (CVMV) and a partial resistance to potato virus Y pathotype 1,2 (PVY 1,2) were found in an Indian pepper line, Perennial. In the doubled haploid (DH) progeny from the F₁ of a cross Perennial by Yolo Wonder, resistance to CVMV was confered by two independent genes, one with a clear dominant effect. Resistance to PVY and potyvirus E was quantitatively expressed and controlled by several recessive genetic factors. Genetic analysis showed that fewer resistance factors were necessary to explain resistance to PVY (0) and potyvirus E than resistance to PVY(1,2). Genetic correlations between resistances to the different potyviruses in the DH progeny showed that most of genetic factors involved in PVY(0) resistance appear to be also involved in potyvirus E resistance, and some of these polyvalent factors may be also involved in PVY(1,2) resistance but, in this case, additional specific genes were necessary. One of the two CVMV resistance genes seems to be implicated in potyvirus E resistance. Thus, the polygenic resistance of Perennial to these potyviruses was due both to polyvalent genetic factors, i.e. factors that apparently interact with several viruses, and strain-specific genetic factors.     

Inhibition of Pepper mild mottle virus with Commercial Cellulases

Pepper mild mottle virus (PMMoV) is one of the most important pathogens of pepper crops (Capsicum annuum L.) worldwide. We have found that commercial cellulases from Trichoderma reesei and Trichoderma viride strongly inhibit PMMoV infection of plants. When purified PMMoV was mixed with the cellulases, it greatly lost the capacity to induce local lesions on leaves of Nicotiana glutinosa. Pretreatment of sweet pepper leaves (C. annuum L.) with cellulase solutions before PMMoV inoculation greatly reduced the number of infected plants. These effects were superior to the effects of known viral inhibitors such as skim milk or Lentinula edodes culture filtrates. Although the mechanism of inhibition remains unknown, this is the first report of antiviral activity ascribed to commercially available cellulases.