Occurrence, Distribution and Relative Importance of Viruses Infecting Hot Pepper and Tomato in the Major Growing Areas of Ethiopia

Hot pepper and tomato fields in the main growing areas in the Rift Valley and the west of Ethiopia were surveyed for virus infections in 1994. A total of 286 samples from hot pepper and 222 samples from tomato plants and associated Datura stramonium L. and Nicandra physalodes Gaertn. weeds with symptoms suggestive of virus infections were collected and analysed using electron microscopy, serology and test plant reactions. Potato virus Y (PVY), Ethiopian pepper mottle virus (EPMV), pepper veinal mottle virus (PVMV) and tomato mosaic virus (ToMV) were detected in hot pepper samples while tomato samples were shown to be infected with tomato mild mottle virus (TMMV), PVY and ToMV. The most widespread and predominant viruses which also occurred frequently in mixed infections were PVY and EPMV in hot pepper and PVY and TMMV in tomato. TMMV was also found in many samples of D. stramonium and N. physalodes. ToMV was identified in only few samples from both crops in the Rift Valley by its characteristic particle morphology, serological properties and symptomatology. PVMV was found in hot pepper samples only from western Ethiopia, but no natural infection of tomato with this virus was revealed. This is the first report on the natural occurrence of TMMV in tomato, D. stramonium and N. physalodes, as well as of ToMV in hot pepper and tomato in Ethiopia.     

Molecular characterisation and relative incidence of bean‐ and soybean‐infecting begomoviruses in northwestern Argentina

Recent studies identified three begomoviruses infecting soybean and bean crops in northwestern (NW) Argentina, bean golden mosaic virus (BGMV), a virus with high capsid protein identity with Sida mottle virus (SiMoV) and a possible new viral species (isolate A). Analysis of complete DNA‐A sequences confirmed that isolate A represents a new viral species for which the authors propose the name soybean blistering mosaic virus (SbBMV), whereas the SiMoV‐like virus is actually an isolate of tomato yellow spot virus (ToYSV). Molecular hybridisation‐based detection of the three begomoviruses was accomplished using a general probe obtained by mixing full‐length DNA‐A clones of the three begomoviruses and specific probes comprising part of the common region of each viral genome. These probes were used to test samples collected in NW Argentina from 2004 through 2007. Fifty‐three percent of the bean samples were infected with BGMV, 11.5% with ToYSV and 9% with SbBMV. For soybean samples, 33% were infected with SbBMV and 18% with ToYSV. BGMV was not detected in soybean. ToYSV was also detected in the wild species Abutilon theophrasti.     

Peanut yellow spot virus: A distinct tospovirus species based on serology and nucleic acid hybridisation

Nucleocapsids of peanut yellow spot virus (PYSV), purified from peanut (= groundnut) plant tissue, contained a protein with a molecular mass of 29 kDa. In ELISA and immuno-blot analysis the virus did not react with tomato spotted wilt virus (TSWV), Impatiens necrotic spot virus (INSV) and peanut bud necrosis virus (PBNV) antisera. PYSV contained three RNA species, a large (L) RNA (c.8900 nucleotides), a medium (M) RNA (c.4800 nucleotides) and a small (S) RNA (c.3000 nucleotides), similar to other tospoviruses. In addition, a fourth RNA species of approximately 1800 nucleotides was also present in purified preparations. Hybridisation analysis under high stringency conditions revealed no hybridisation between PYSV RNAs and cDNA probes representing the nucleocapsid (N) gene, the glycoprotein (GP) gene and the 3' half of the RNA polymerase gene of PBNV. PYSV genomic RNAs also failed to hybridise with cDNA probes from the GP genes of TSWV and INSV. In reciprocal tests, the cDNA clones of PYSV S and M RNAs did not hybridise with any of the PBNV RNAs. Based on the absence of serological relationships between PYSV and PBNV, TSWV and INSV and lack of nucleotide homology based on hybridisation studies between the PYSV RNAs and cDNA clones from PBNV, TSWV and INSV, PYSV should be considered as a distinct species of the genus Tospovirus under a new serogroup, putatively designated ‘V’.     

Integrated biological control of bacterial speck and spot of tomato under field conditions using foliar biological control agents and plant growth-promoting rhizobacteria

Integration of foliar bacterial biological control agents and plant growth promoting rhizobacteria (PGPR) was investigated to determine whether biological control of bacterial speck of tomato, caused by Pseudomonas syringae pv. tomato, and bacterial spot of tomato, caused by Xanthomonas campestris pv. vesicatoria and Xanthomonas vesicatoria, could be improved. Three foliar biological control agents and two selected PGPR strains were employed in pairwise combinations. The foliar biological control agents had previously demonstrated moderate control of bacterial speck or bacterial spot when applied as foliar sprays. The PGPR strains were selected in this study based on their capacity to induce resistance against bacterial speck when applied as seed and soil treatments in the greenhouse. Field trials were conducted in Alabama, Florida, and California for evaluation of the efficacy in control of bacterial speck and in Alabama and Florida for control of bacterial spot. The foliar biological control agent P. syringae strain Cit7 was the most effective of the three foliar biological control agents, providing significant suppression of bacterial speck in all field trials and bacterial spot in two out of three field trials. When applied as a seed treatment and soil drench, PGPR strain Pseudomonas fluorescens 89B-61 significantly reduced foliar severity of bacterial speck in the field trial in California and in three of six disease ratings in the field trials in Alabama. PGPR strains 89B-61 and Bacillus pumilus SE34 both provided significant suppression of bacterial spot in the two field trials conducted in Alabama. Combined use of foliar biological control agent Cit7 and PGPR strain 89B-61 provided significant control of bacterial speck and spot of tomato in each trial. In one field trial, control was enhanced significantly with combined biological control agents compared to single agent inoculations. These results suggest that some PGPR strains may induce plant resistance under field conditions, providing effective suppression of bacterial speck and spot of tomato, and that there may be some benefit to the integration of rhizosphere-applied PGPR and foliar-applied biological control agents.     

Constitutive Expression of Interferon-Induced Human MxA Protein in Transgenic Tobacco Plants Does not Confer Resistance to a Variety of RNA Viruses

MxA is a key component in the interferon-induced antiviral defense in humans. After viral infections, MxA is rapidly induced and accumulates in the cytoplasm. The multiplication of many RNA viruses,including all bunyaviruses tested so far, is inhibited by MxA. These findings prompted us to express MxA in plants in an attempt to create resistance to tospoviruses. Here, we report the generation of transgenic tobacco plants that constitutively express MxA under the control of the 35S cauliflower mosaic virus promotor. Northern and western blot analysis confirmed the expression of MxA in several transgenic plant lines. MxA expression had no obvious detrimental effects on plant growth and fertility. However, challenge experiments with tomato spotted wilt virus, tomato chlorotic spot virus, and groundnut ringspot virus revealed no increased resistance of MxA-transgenic tobacco plants to tospovirus infections. Neither was the multiplicationof tobacco mosaic virus, cucumber mosaic virus and potato virus Y inhibited in MxA-transgenic plants. The results indicate that the expression of human MxA alone does not enhance virus resistance in planta.     

A comparison of the effectiveness of the four British virus vector species of Longidorus and Xiphinema

The frequency with which the four virus-vector species of longidoroid nematodes occurring in Britain transmitted their associated plant viruses were compared in a series of experiments using a standard procedure. In these tests Xiphinema diversicaudatum proved an effective vector of British isolates of arabis mosaic virus and strawberry latent ringspot virus and Longidorus attenuatus of an isolate of tomato black ring virus from England. In comparison, isolates of raspberry ringspot virus and tomato blackring virus from Scotland and of raspberry ringspot virus from England were transmitted much less readily by their respective vectors, L. elongatus and L. macrosoma. These differences in ability to transmit virus were not related to differences in feeding access on the virus source- or bait-plants, in the extent to which virus was retained within the nematode feeding apparatus or in the frequency with which virus was recovered from Longidorus in concurrent slash tests. Three Scottish isolates of raspberry ringspot and tomato black ring viruses were transmitted equally infrequently by two populations of L. elongatus and the frequency with which virus was transmitted was not greatly increased when the species of source or bait plants was changed.     

Transgenic tomato plants expressing the <Emphasis Type="Italic">Arabidopsis NPR1</Emphasis> gene display enhanced resistance to a spectrum of fungal and bacterial diseases

Development of effective disease-resistance to a broad-range of pathogens in crops usually requires tremendous resources and effort when traditional breeding approaches are taken. Genetic engineering of disease-resistance in crops has become popular and valuable in terms of cost and efficacy. Due to long-lasting and broad-spectrum of effectiveness against pathogens, employment of systemic acquired resistance (SAR) for the genetic engineering of crop disease-resistance is of particular interest. In this report, we explored the potential of using SAR-related genes for the genetic engineering of enhanced resistance to multiple diseases in tomato. The Arabidopsis NPR1 (nonexpresser of PR genes) gene was introduced into a tomato cultivar, which possesses heat-tolerance and resistance to tomato mosaic virus (ToMV). The transgenic lines expressing NPR1 were normal as regards overall morphology and horticultural traits for at least four generations. Disease screens against eight important tropical diseases revealed that, in addition to the innate ToMV-resistance, the tested transgenic lines conferred significant level of enhanced resistance to bacterial wilt (BW) and Fusarium wilt (FW), and moderate degree of enhanced resistance to gray leaf spot (GLS) and bacterial spot (BS). Transgenic lines that accumulated higher levels of NPR1 proteins exhibited higher levels and a broader spectrum of enhanced resistance to the diseases, and enhanced disease-resistance was stably inherited. The spectrum and degree of these NPR1-transgenic lines are more significant compared to that of transgenic tomatoes reported to date. These transgenic lines may be further explored as future tomato stocks, aiming at building up resistance to a broader spectrum of diseases.     

Biological control of bacterial spot of tomato under field conditions at several locations in North America

Following the relatively successful biological control of bacterial speck of tomato under field conditions at several locations (Phytopathology 92 (2002) 1284), similar selection and testing strategies were employed in an effort to isolate an effective biological control agent for bacterial spot of tomato. Fifty potential biological control agents were isolated from tomato foliage in Alabama (AL) and Florida (FL) and tested under greenhouse conditions in AL for the ability to reduce the foliar severity of bacterial spot of tomato (Lycopersicon esculentum), which is caused by either Xanthomonas campestris pv. vesicatoria or Xanthomonas vesicatoria. Three pseudomonads that provided protection against bacterial speck also were included in the tests. The strains which were most efficacious (i.e., high mean percentage reduction) and consistent (i.e., low standard deviation) in reducing bacterial spot severity in repeated greenhouse experiments were selected for field experiments conducted over the period 1996–1998. Among these strains were Cellulomonas turbata BT1, which provided the highest mean reduction in disease severity [45.2% (SD = 21.0)], and Pseudomonas syringae Cit7 [36.4% (SD = 12.2)], which was the most consistent. Field experiments were conducted in Shorter, AL; Bradenton and Sanford, FL; Clinton, North Carolina; Wooster, Ohio; and London, Ontario, Canada. The highest mean reductions in severity of bacterial spot on foliage, averaged across all locations, were provided by P. syringae Cit7 [28.9% (SD = 11.6)] and Pseudomonas putida B56 [23.1% (SD = 7.4)]. The efficacy and consistency of P. syringae Cit7 against bacterial spot were very similar to those achieved against bacterial speck [28.3% (SD = 12.7)] (Phytopathology 92 (2002) 1284). Unfortunately, neither the bacterial strains nor the standard copper bactericides consistently reduced disease incidence on fruit.     

HEAT-THERAPY OF VIRUS-INFECTED PLANTS

Virus-free plants were produced from parents systemically infected with the following five viruses: tomato bushy stunt, carnation ring spot, cucumber mosaic, tomato aspermy and Abutilon variegation. The leaves formed while the infected plants were kept at 36°C. were free from symptoms, and test plants inoculated from these remained uninfected. When cuttings were taken from the infected plants at the end of the treatment most grew into healthy plants. The treated plants themselves usually developed symptoms after varying lengths of time at 20°C, but some that before treatment were infected with tomato aspermy, cucumber mosaic or Abutilon variegation viruses, remained permanently healthy.
The same method failed to cure plants infected with tomato spotted wilt, potato virus X and tobacco mosaic virus, although it decreased their virus content. Heat-therapy seems not to be correlated with the thermal inactivation end point of the virus in vitro.     

Induction of defence-related enzymes in tomato (Solanum lycopersicum) plants treated with Bacillus subtilis CBR05 against Xanthomonas campestris pv. vesicatoria

Bacterial spot disease caused by Xanthomonas campestris pv. vesicatoria is one of the most important destructive diseases of tomato in many parts of the agricultural world. Therefore, the present study aims to determine the effects of Bacillus subtilis CBR05 inoculation on bacterial spot disease severity and the induction of defence-related enzymes response in tomato. Tomato leaves were evaluated to determine the activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and polyphenol oxidase (PPO)) and the content of malondialdehyde (MDA). A reduction in bacterial spot severity was observed in plants inoculated with B. subtilis, compared with those of uninoculated controls. A significant increase in SOD, CAT, POD, and PPO activities was observed in plants treated with B. subtilis after 24?h inoculation compared with non-inoculated pathogen control and mock-inoculated controls. Moreover, the MDA content was induced by pathogen infection, and its amount in B. subtilis inoculated plants was significantly lower than that in pathogen control. Our results suggest that early increases in antioxidant enzymes and the reduction in MDA content with B. subtilis inoculation may play a pivotal role in mitigating oxidative stress, thereby induced systemic resistance against bacterial spot disease in tomato.     

Induced systemic resistance to tomato leaf curl virus and increased yield in tomato by plant growth promoting rhizobacteria under field conditions

The talc-based formulation of two Pseudomonas fluorescens strains (Pf1 and VPT10) and its mixture (with and without chitin) were tested against tomato leaf curl virus in tomato under greenhouse and field conditions. The mean percentage of tomato leaf curl virus infected plants were significantly lower (25%) with less symptom severity and delayed symptom expression up to nine additional days in Pseudomonas with chitin (VPT10 + chitin) treated tomato plants compared to non-bacterised control plants upon challenge inoculation with tomato leaf curl virus. Tomato leaf curl virus was partially purified and antiserum was developed. Using the antiserum the tomato leaf curl virus was detected in symptomatic leaves and in whitefly vector through direct antigen coating enzyme linked immunosorbent assay which revealed the low virus titre in Pseudomonas treated plants (VPT10 + chitin) and insect vector compared to untreated tomato plants. The results indicate the potentiality of plant growth promoting rhizobacteria strains and talc-powder formulations in the effective management of this tomato leaf curl virus in tomato under field conditions.     

Natural infection of sugar beet with tomato bushy stunt virus

A virus transmissible toChenopodium quinoa was isolated from leaves of sugar beet showing large chlorotic ring spots and line pattern. The virus was serologically unrelated to tobacco necrosis virus and tomato black ring virus or to its beet ringspot strain either. A positive result was obtained with antiserum against tomato bushy stunt virus. Reactions of herbaceous indicators and properties of the virus in crude sap were in accordance with the serological diagnosis. A survey of natural hosts of tomato bushy stunt virus demonstrated recently by the authors is given.     

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.     

Hyaluronic acid of Streptococcus sp. as a potent elicitor for induction of systemic resistance against plant diseases

The potential of hyaluronic acid (HA) in inducing systemic resistance to cucumber, tomato and pepper was tested in planta. In the study, HA was found to be a potent agent for suppressing disease caused by Cucumber Mosaic Virus (CMV) (in pepper), Pseudomonas syringae pv. tomato (tomato speck disease), Xanthomonas axonopodis pv. vesicatoria (tomato spot disease), Pseudomonas syringae pv. lachrymans (cucumber angular leaf spot), and Colletotrichum orbiculare (cucumber anthracnose). Disease control was obtained with spraying, injection and drenching of plants with HA. HA did not exhibit direct antimicrobial action against the pathogens tested. Studies carried out in transgenic tobacco indicated that defense genes PR 1a and PDF 1.2 were activated upon treatment with HA, demonstrating salicylic acid (SA) and jasmonic acid (JA) pathways getting activated during defense. Further work is warranted to evaluate the use of HA-mediated disease suppression in crop plants.     

Effects of virus genotype and temperature on seed transmission of nepoviruses

Transmission of different nepoviruses through chickweed (Stellaria media) seed was differently affected by ambient temperature during seed production. Raspberry ringspot and tomato black ring (Scottish isolate) viruses were similarly and frequently transmitted at 14 , 18 and 22 ^oC, whereas arabis mosaic virus was transmitted most frequently at 14 ^oC, and strawberry latent ringspot and tomato black ring (German isolate) viruses at 22 ^oC. When infected by seed-borne nepoviruses, seedlings of S. media and other species were symptomless at 15–25 ^oC, and the viruses were therefore detected by inoculating sap to Chenopodium quinoa indicator plants. However, typical symptoms of arabis mosaic and tomato black ring viruses were induced by growing Nicotiana tabacum, N. clevelandii and C. quinoa seedlings infected with seed-borne virus at 33–37 ^oC during the third and fourth weeks after sowing, preceded and followed by periods at 15–25 ^oC. The proportion of N. tabacum seedlings developing symptoms was the same as that of untreated seedlings yielding sap-transmissible virus. Seed transmissibility of pseudo-recombinant isolates of raspberry ringspot and tomato black ring viruses, containing RNA-i from one virus strain and RNA-2 from another strain, depended greatly on the transmissibility of the strain contributing RNA-i. The source of RNA-2 had an additional but smaller influence. The satellite RNA (RNA-3) of tomato black ring virus was seed-transmitted in S. media and its occurrence in cultures did not affect the frequency of transmission of the virus. Results of testing the infectivity of extracts of seed from infected mother plants suggested that failure of seed transmission reflected failure to become established in the seed, not subsequent inactivation. Whereas seed transmissibility of raspberry ringspot virus is primarily dependent on information carried in RNA-i, transmissibility by nematode vectors, another property of major ecological importance, is determined by RNA-2. In the field, selection pressures presumably can act independently on the two parts of the genome but evidence was also obtained of selection for mutual compatibility of RNA-i and RNA-2.     

Temporal Dynamics of Tomato Severe Rugose Virus and Bemisia tabaci in Tomato Fields in São Paulo,Brazil

Temporal progress of a begomovirus disease in tomato fields and the abundance of its whitefly vector, Bemisia tabaci biotype B, were evaluated during three consecutive tomato plantings in the municipality of Sumaré, state of São Paulo, Brazil, in 2006 and 2007. The incidence of symptomatic plants and the number of adult whiteflies were weekly monitored on experimental plots randomly chosen in tomato commercial fields. Tomato severe rugose virus (ToSRV) was identified as the causal agent of the disease, and its relationships with other Brazilian begomoviruses was confirmed by partial and complete nucleotide sequencing of the viral genome. The disease temporal progress was analysed by fitting different models to disease incidence. The monomolecular model showed the best fit, which is consistent with a predominant role of primary spread in the epidemiology of ToSRV. A higher number of adult whiteflies were observed at the borders of the plots, also suggesting primary spread of ToSRV from external sources of inoculum, which might be represented by weeds and volunteer tomato‐infected plants. In Brazil, since 2004, there is a legislative measure that mandates, for some regions of processing tomato plantings, a 2‐month crop‐free period during the year. Based on our results, we suggest the extension of this measure to all tomato‐producing regions, including fresh market tomato. We also suggest that growers emphasize the elimination of old plants from harvested fields that can serve as virus reservoirs several weeks prior to new plantings and weeds nearby the fields to limit the primary spread of ToSRV.     

The contribution of tomato and alternative host plants to tomato leaf curl virus inoculum pressure in different areas of South India

Indian tomato leaf curl virus (ToLCV) (Geminiviridae: Sub-group III) was detected both in field-collected and laboratory-reared B. tabaci using a triple-antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA). ToLCV was detected in six of the 10 group samples of field collected B. tabaci. ToLCV was also identified in 13 weed species commonly found in Karnataka, both by symptom expression and TAS-ELISA. ToLCV from c. 61% of infected plants was transmitted successfully to tomato by B. tabaci. Tomato plots were planted at three locations on the University of Agricultural Sciences Campus, Bangalore. Indian tomato leaf curl virus disease (ToLCVD) incidence increased most rapidly when the tomato plot was situated adjacent to an older ToLCVD-infected tomato field. When the plots were positioned in a dryland or a wetland area, at least 500 m away from any infected tomato fields, the ToLCVD incidence increased less rapidly, although in all sites it was 100% by 11 wk after transplanting. The numbers of B. tabaci caught on yellow traps in all sites increased during weeks 1–3 after transplanting and thereafter remained at between 10–15 adults trap^-1 24 h^_1. Adult numbers recorded on tomato plants by direct counts remained approximately constant at 2–4 adults plant“”¹. Tomato fields were planted in three taluks (administrative areas) of Karnataka, that had different current and previous histories of tomato production. ToLCVD incidence increased most and least rapidly, respectively, in Kolar taluk where tomato is grown continuously and Doddaballapur tuluk where tomato was grown in the area for the first time. In Malur tuluk, where tomato was grown discontinuously (once a year), the incidence of ToLCVD increased at an intermediate rate. Weed host-plant species growing near the experimental sites had averages of between 1.5–10.0 B. tabaci nymphs per plant, whereas the tomato plants had only 0.3 nymphs per plant. The percentage parasitism of B. tabaci nymphs on tomato and weed species, respectively, was 0.7% and 2–6%. Nymphs and pupae were parasitised by an Encarsia sp. and Eretmocerus mundus Mercet. The relevance and implications of these findings for the epidemiology and management of ToLCVD in Karnataka State, South India is discussed.     

Detection of three whitefly-transmitted geminiviruses occurring in Europe by tests with heterologous monoclonal antibodies

Selected monoclonal antibodies (MAbs), prepared to particles of African cassava mosaic or Indian cassava mosaic geminiviruses, detected three geminiviruses that occur in Europe: abutilon mosaic virus in Abutilon pictum ‘Thompsonii’, tobacco leaf curl virus in Lonicera japonica var. aureo-reticulata and tomato yellow leaf curl virus in Lycopersicon esculentum. All three viruses were detected in indirect ELISA by MAbs SCR 17 and SCR 20 but they were differentiated by their reactions with SCR 18 and SCR 23. Tobacco leaf curl virus was detected only when reducing agents were included in the leaf extraction medium. Inclusion of sodium sulphite slightly improved detection of tomato yellow leaf curl virus but reducing agents were not needed for detection of abutilon mosaic virus.