Virus Diseases of Pepper (Capsicum annuum L.) in Ethiopia

Viruses occurring in hot pepper (Capsicum annuum L.) in Ethiopia during 1984–1986 were identified and their symptomatology, host range, serology, morphology and transmission described. Potyviruses were found to cause severe infections of pepper wherever the crop was observed. Disease incidence and yield loss in most surveyed areas were estimated at 40–100 % and 15–50 %, respectively. Besides potato Y and pepper veinal mottle viruses, a new potyvirus with distinctive properties was found widely distributed in Shewa and Welega Provinces. The virus, tentatively designated Ethiopian pepper mottle virus (EPMV), was shown to have an unusually narrow experimental host range and filamentous particles about 700–750 nm in length, and to be nonpersis-tently transmitted by aphids but not transmissible through pepper seeds. Cucumber mosaic virus was detected in some locations, but appeared to be of minor significance.     

Studies on plant growth-regulating substances LV. The Plant growth-promoting properties of some β-aryl-propionic acids

Severe diseases of pepper (Capsicum annuum), tomato (Lycopersicon esculentum), eggplant (Solanum melongena) and tomato eggplant (Solanum integrifolium) in West Africa were induced by pepper veinal mottle virus (PVMV). Five selected virus isolates were serologically similar and readily transmissible by aphids in the non-persistent manner, but they differed in host range and/or symptoms induced in some susceptible species. One isolate from eggplant failed to infect pepper, Chenopodium quinoa and C. amaranticolor, and induced only local infections in tomato. An isolate from tomato failed to infect eggplant, and an isolate from tomato eggplant induced severe stunting in Physalis floridana. The type strain, like the isolate from tomato, failed to infect Nicotiana tabacum systemically, but each caused severe systemic leaf and stem necrosis in tomato. None of the isolates infected S. melongena cv. Long Purple, suggesting that PVMV might be controlled in this and perhaps other crop species by the use of immune or tolerant cultivars. All five isolates were serologically related to potato virus Y and some to six of 12 other potyviruses.     

The potential use of a viral coat protein gene as a transgene screening marker and multiple virus resistance of pepper plants coexpressing coat proteins of cucumber mosaic virus and tomato mosaic virus

Transgenic pepper plants coexpressing coat proteins (CPs) of cucumber mosaic virus (CMV-Kor) and tomato mosaic virus (ToMV) were produced by Agrobacterium-mediated transformation. To facilitate selection for positive transformants in transgenic peppers carrying an L gene, we developed a simple and effective screening procedure using hypersensitive response upon ToMV challenge inoculation. In this procedure, positive transformants could be clearly differentiated from the nontransformed plants. Transgenic pepper plants expressing the CP genes of both viruses were tested for resistance against CMV-Kor and pepper mild mottle virus (PMMV). In most transgenic plants, viral propagation was substantially retarded when compared to the nontransgenic plants. These experiments demonstrate that our transgenic pepper plants might be a useful marker system for the transgene screening and useful for classical breeding programs of developing virus resistant hot pepper plants.     

Detection of Tobacco mosaic virus and Tomato mosaic virus in pepper and tomato by multiplex RT-PCR

Aims: To develop a highly sensitive and rapid protocol for simultaneous detection and differentiation of Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) in pepper and tomato. In this study, we use the multiplex PCR technique to detect dual infection of these two viruses. Methods and Results: A multiplex RT–PCR method consisting of one‐tube reaction with two primer pairs targeted to replicase genes was developed to simultaneously detect TMV and ToMV in seed samples of pepper and tomato. Specific primers were designed from conserved regions of each of the virus genomes, and their specificity was confirmed by sequencing PCR products. RT–PCR detected up to 10^?6 dilution of total RNA extracted from infected leaves. Multiplex RT–PCR revealed the presence of both TMV and ToMV in three of 18 seed samples of tomato and one of 18 seed samples of pepper. Conclusions: The multiplex PCR assay was a cost effective, quick diagnostic technique, which was helpful in differentiating TMV and ToMV accurately. Significance and Impact of the Study: The multiplex PCR assay described in this study is a valuable tool for plant pathology and basic research studies. This method may facilitate better recognition and distinction of TMV and ToMV in both pepper and tomato.     

Disease intensity of some tomato viroses in Serbia

In this paper we present the data on the disease intensity of the tomato plants grown in glass and plastic-houses, and in the open field. The infection was caused by the following viruses: Tomato mosaic virus (ToMV), Tobacco mosaic virus (TMV), Tomato spotted wilt virus (TSWV), Alfalfa mosaic virus (AMV), Potato virus X (PVX), Potato virus Y (PVY), Tomato black ring virus (TBRV), Tomato ringspot virus (ToRSV), Tomato aspermy virus (TAV), and Cucumber mosaic virus (CMV). These viruses represented most frequent tomato pathogens in Serbia. According to the obtained results, it could be concluded that 92.94% of the tested tomato plants grown in glass and plastic-houses, and 89.82% grown in the open field were infected by one of the above viruses. Most of the plant samples were infected by two or more viruses. The most frequent viruses — tomato pathogens in Serbia were ToMV, PVY and TMV.     

Detection of tobacco mosaic virus and tomato mosaic virus in pepper seeds by enzyme linked immunosorbent assay (ELISA)

Pepper seed samples were tested for the infection of tobacco mosaic virus (TMV) and tomato mosaic virus (ToMV) by enzyme linked immunosorbent assay (ELISA). Out of 26 pepper seed samples tested, 17 were infected with TMV and ToMV in ELISA. About 34.7% of pepper seed samples were found to be healthy. Infections of TMV or ToMV were recorded to be 61.53% and 11.5%, respectively of the total tested seed samples.     

Nucleotide sequence of Chinese rape mosaic virus (oilseed rape mosaic virus), a crucifer tobamovirus infectious on Arabidopsis thaliana

The complete nucleotide sequence of Chinese rape mosaic virus has been determined. The virus is a member of the tobamovirus genus of plant virus and is able to infect Arabidopsis thaliana (L.) Heynh systemically. The analysis of the sequence shows a gene array that seems to be characteristic of crucifer tobamoviruses and which is slightly different from the one most frequently found in tobamoviruses. Based on gene organization and on comparisons of sequence homologies between members of the tobamoviruses, a clustering of crucifer tobamoviruses is proposed that groups the presently known crucifer tobamovirus into two viruses with two strains each. A name change of Chinese rape mosaic virus to oilseed rape mosaic virus is proposed.Abbreviations 2-ME 2-mercaptoethanol - EDTA ethylenediaminetetraacetic acid - SDS sodium dodecyl sulfate - UTR untranslated region - MP movement protein - CP capsid protein - CRMV Chinese rape mosaic virus - TVCV turnip vein clearing virus - PaMMV paprika mild mottle virus - PMMV-I pepper mild mottle virus (Italian isolate) - PMMV-S pepper mild mottle virus (Spanish isolate) - ToMV tomato mosaic virus - TMV tobacco mosaic virus - TMGMV tobacco mild green mosaic virus - ORSV odontoglossum ringspot virus - SHMV sunn hemp mosaic virus - CGMMV cucumber green mottle mosaic virus - ORMV oilseed rape mosaic virus     

Beetle, contact and potato true seed transmission of Andean potato latent virus

A species of the flea beetle (Epitrix sp.) transmitted Andean potato latent virus (APLV) from Datura stramonium to D. stramonium, Nicandra physalodes and potato, and from N. physalodes to N. physalodes and D. stramonium. Although the acquisition and inoculation feeding times used were varied, this transmission always occurred at low efficiency. By contrast, APLV was very readily transmitted in tests in which infected plants of D. stramonium, N. physalodes, Nicotiana bigelovii and potato were brushed against healthy plants of one or more of these species. A low level of seed transmission of APLV occurred in potato.     

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.     

Systemic Infection of Tobacco by Pepper Veinal Mottle Potyvirus (PVMV) Depends on the Presence of Potato Virus Y (PVY)

In single inoculations, both PVY and PVMV replicated in inoculated leaves of Nicotiana tabacum cv. ‘Xanthi nc’ plants, but only PVY infected the tobacco plants systemically, whereas PVMV caused localized infection. A mixed infection by the PVY-To72 and PVMV-type strains was experimentally realized in ‘Xanthi nc’ plants. In the presence of PVY, PVMV migrated systemically into the upper leaves of the tobacco plant, as was proved by back inoculation. It would appear that in tobacco, PVY acts as a “helper” virus, providing PVMV with the necessary component factor for migration. In extracts from the co–infected leaves. Immune Electron Microscopy (IEM) revealed phenotypic mixed particles which contained a mixture of coat proteins of PVY and PVMV. The role of the structural and functional interactions between the two viruses, which enable PVMV to migrate systemically in tobacco plants, is discussed.     

Incidence and epidemiology of Pepper veinal mottle virus (PVMV), a Potyvirus disease of pepper

There have been various reports about the devastating effect of Pepper veinal mottle virus (PVMV), a Potyvirus disease of pepper in Nigeria contributing to its low yield and reduced fruit quality leading to great economic loss. Different strains of the virus have been identified and characterised over the years and the disease incidence, severity and aphid vector distribution across agro-ecological zones studied. Different cultural management techniques have been tried and found to be effective with varying degree of success, and these included the use of organic manures, intercropping with tall companion crops, time and date of planting and the use of tolerant/resistant varieties. Integrated pest management techniques for PVMV disease have been found to be very effective.     

Serodiagnosis of Pepper Veinal Mottle Virus in West Africa using Specific Monoclonal Antibodies in DAS-ELISA

In order to develop a reliable diagnostic test for pepper veinal mottle virus (PVMV), a panel of monoclonal antibodies (mAbs), raised to a Nigerian isolate (PVMW-Ni), was evaluated by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) on infected pepper samples from seven West African countries. PVMV from different geographical locations was serologically very homogeneous and no significant epitope differences were detected even with a very sensitive double-mAb sandwich ELISA. This ELISA format was optimized with mAb 6C12 and its application in a diagnostic test resulted in the identification of PVMV in Burkina Faso, Togo and Sénégal where it had not been previously reported.     

Monitoring of co-infection virus and virus-like naturally in sweet pepper plant

Eight sweet pepper plant samples showing viral and viral like symptoms were collected from open field and used for detecting viral infections through biological, serological and biochemical methods. DAS-ELISA, DBIA and TPIA have relative effectiveness for detecting parenchymal viruses (CMV, TMV and PVY) and vascular virus (TYLCV), and the DAS-ELISA and TPIA are found more efficient (87.5%) than DBIA (78.1%). The examined leaf samples were found co-infected with different mixed types of viruses including (CMV, TMV, PVY and TYLCV), (CMV, PVY and TYLCV), (TMV, PVY and TYLCV) and (TMV and TYLCV) that enhanced different degrees of severe external symptoms. There are 2 out of 8 samples infected with Phytoplasma sp. by Diene’s stain and PCR using generated 16S rDNA gene primer with expected amplicon size of 680?bp. The co-infections with various viruses and phytoplasma has 12.5% frequency that reduced the levels of protein content, peroxidase and polyphenol oxidase activity quantitatively and qualitatively in 2 samples in comparison with other mixed categories. The sweet pepper plant can be considered as a reservoir for parenchymal and vascular viruses and Phytoplasma sp. due to the synergistic and antagonistic effects causing unusual and unpredictable biological and epidemiological, viral and viral-like via host biochemical effects.     

Interactions between pepper ringspot virus and cylindrical inclusions of two potyviruses

Immunoelectron microscopy showed that cylindrical inclusions (CI) of the potyvirus potato virus Y (PVY) bound in addition to their homologous virions those of a co-infecting rod-shaped virus, pepper ringspot virus (PRV), in infected Nicotiana benthamiana leaf cells. The latter virus does not code for cylindrical inclusions and is cl assified as a Tobravirus. Virions of PRV were scattered throughout the cell cytoplasm and not associated with mitochondria in PVY + PRV double infections. Binding of PRV to mitochondria was disrupted in PVY + PRV infected cells. In double infections with a second potyvirus, tobacco etch virus (TEV), and PRV in N. benthamiana cells, TEV-CI bound homologous TEV virions but did not bind PRV. In contrast to PVY + PRV infections, virions of PRV attached end-on to mitochondrial limiting membranes in PRV-only and in TEV + PRV double infections. The results are interpreted to mean that there are differences in the PRV virion binding sites of PVY-CI and TEV-CI. In previous reports, potyviral CI have been nondiscriminating in binding virions or capsid proteins of other co-infecting rod-shaped viruses.     

Concurrent evolution of resistance and tolerance to potato virus Y in Capsicum annuum revealed by genome-wide association

We performed a genome-wide association study of pepper (Capsicum annuum) tolerance to potato virus Y (PVY). For 254 pepper accessions, we estimated the tolerance to PVY as the coefficient of regression of the fresh weight (or height) of PVY-infected and mock-inoculated plants against within-plant virus load. Small (strongly negative) coefficients of regression indicate low tolerance because plant biomass or growth decreases sharply as virus load increases. The tolerance level varied largely, with some pepper accessions showing no symptoms or fairly mild mosaics, whereas about half (48%) of the accessions showed necrotic symptoms. We found two adjacent single-nucleotide polymorphisms (SNPs) at one extremity of chromosome 9 that were significantly associated with tolerance to PVY. Similarly, in three biparental pepper progenies, we showed that the induction of necrosis on PVY systemic infection segregated as a monogenic trait determined by a locus on chromosome 9. Our results also demonstrate the existence of a negative correlation between resistance and tolerance among the cultivated pepper accessions at both the phenotypic and genetic levels. By comparing the distributions of the tolerance-associated SNP alleles and previously identified PVY resistance-associated SNP alleles, we showed that cultivated pepper accessions possess favourable alleles for both resistance and tolerance less frequently than expected under random associations, while the minority of wild pepper accessions frequently combined resistance and tolerance alleles. This divergent evolution of PVY resistance and tolerance could be related to pepper domestication or farmer's selection.     

Pepper veinal mottle virus-a new member of the potato virus Y group from peppers (Capsicum annuum L. and C. frutescens L.) in Ghana

Pepper veinal mottle virus (PVMV), a previously undescribed virus widespread in Capsicum annuum and C. frutescens in the Eastern Region of Ghana, is acquired and inoculated in 2 min feeding periods by aphids (Myzus persicae and Aphis gossypii); it is transmissible by inoculation of sap to eleven of fifteen Solanaceae and to five of forty-six other species within three of seventeen other families. The virus was propagated in Nicotiana clevelandii and Petunia hybrida, and assayed in Chenopodium quinoa, C. amaranticolor and C. murale. Sap from Capsicum annuum was infective after dilution to 10^-3 but not 10^-4, after 10 min at 55 but not 60^oC, and after 7 but not 8 days at 25^oC. Lyophilized sap from P. hybrida was infective after 6 years in vacuo. Yields of 10–25 mg of virus per kg of leaf tissue were consistently obtained from P. hybrida or N. clevelandii by extracting systemically infected leaves in 0.5 M borate (pH 7.8) containing 0.2% mercaptoethanol and chloroform, followed by repeated precipitation with 50 g polyethylene glycol (M.W. 6000) per l, several cycles of differential centrifugation and centrifugation in sucrose density-gradient columns. Virus preparations had ultraviolet absorption spectra typical of a nucleoprotein containing c. 6% nuclei acid (A 260/280 = 1.25; A 260/246 = 1.27) and contained numerous unaggregated and unbroken filamentous particles c. 770 times 12 nm which sedimented as a single component with a sedimentation coefficient (s^o_20,w) of 155 S. PVMV contained RNA (moles %: G = 24, A = 23, C = 27, U = 26), and a single protein species with a molecular weight of 32000–33000 daltons. PVMV was not serologically related to potato virus Y (three strains), or to twelve other morphologically similar viruses, and seems to be a distinct member of the potato virus Y group. The cryptogram of PVMV is R/(I):*/(6):E/E:S/Ap.     

Accumulation of potato virus Y is enhanced in Solatium brevidens also infected with tobacco mosaic virus or potato spindle tuber viroid

The accumulation of potato virus Y?(PVY?) and potato leaf roll virus (PLRV) was studied in plants of Solanum brevidens co-infected with each of six viruses or a viroid. Virus could not be detected by ELISA in plants of S. brevidens infected solely with PVY. However, accumulation of PVY was increased c. 1000-fold in plants doubly infected with tobacco mosaic virus or potato spindle tuber viroid (PSTVd). PVY titres in doubly infected plants of S. brevidens were between 1% and 0.1% of those found in the PVY-susceptible interspecific Solanum hybrid DTO-33. Double infections of 5. brevidens by PVY and alfalfa mosaic virus or potato viruses M, S, T or X did not significantly enhance PVY accumulation. Accumulation of PLRV was not enhanced in plants co-infected with any of the six viruses or PSTVd.     

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.     

Serological comparison of an Australian isolate of capsicum mosaic virus with capsicum tobamovirus isolates from Europe and America

A New South Wales isolate (Ca) of capsicum mosaic virus was tested against antisera to it and capsicum tobamovirus isolates from the Netherlands (P8, P11), USA (SL), Argentina (FO) and Sicily (PM). The comparison demonstrated that the four viruses Ca, P8, PM and SL are closely related to each other, forming a series of decreasing relationship to Ca in the above order. FO was related to these but insufficiently to be considered part of the group, and P11 was only slightly related to the others. The literature on serology of tobamoviruses in Capsicum spp. was collated and it is suggested that isolates from Sicily (pepper mild mottle), Australia (capsicum mosaic), The Netherlands (P8, P14) and USA (SL) be considered as strains of a virus distinct from both tobacco mosaic and tomato mosaic viruses and that these isolates all be referred to in future as strains of pepper mild mottle virus.