我国南方花生发生一种由蕃茄斑萎病毒引起的新病害

我们于1984和1985年6月上、中旬,在广州市郊、县,湛江市郊以及广西南宁市郊、县,北海市郊和合蒲县等花生产区,调查花生病毒病时,除了发现花生轻斑驳病毒病外,还发现一种新的病毒病害。其症状特征是:病株顶端叶片上出现很多褪绿黄斑或环斑,有的环斑变     

Occurrence and distribution of lettuce mosaic disease in Tehran province from Iran

Lettuce mosaic virus (LMV) Cucumber mosaic virus (CMV) and Tomato spotted wilt virus (TSWV) were identified in fields of Tehran province. In this study 452 leaf samples were collected from the fields throughout the Tehran province during 2002 and 2003. Distribution of Lettuce mosaic virus (LMV), Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV)and Arabis mosaic virus (ArMV) was determined with DAS-ELISA. Percentage of single Infection to LMV. CMV or TSWV was 20.58, 15.93 and 9.96% respectively. Also 15.28% of samples were co- infected with LMV+CMV, 8.19% with LMV+TSMV and 7.74% with CMV+TSWV. 4.65% of samples were Infected to all of these three viruses. LMV was found in 48.69%, CMV in 43.59% and TSWV in 30.54% of samples totally. Therefore LMV is major dominant agent of lettuce mosaic disease in Tehran province. This is the first report of occurrence of TSWV on lettuce in Iran and first report of CMV and LMV in Tehran province.     

Weed species in tomato production and their role as alternate hosts of Tomato spotted wilt virus and its vector Frankliniella occidentalis

Tomato spotted wilt virus (TSWV) is an important plant virus that infects a wide range of hosts including weeds making its management difficult. A survey was undertaken to establish the occurrence of weed species in tomato production systems in Kenya and their role as hosts of TSWV and its vectors. Selected weed species were further evaluated for their reaction to TSWV, transmission efficiency by Frankliniella occidentalis and ability to support thrips reproduction. Of the 43 weed species identified in the field, 29 species had been reported as hosts of TSWV, two were non‐hosts and 11 had no record of their status. Among the more common species, Amaranthus hybridus, Solanum nigrum, Tagetes minuta and Datura stramonium were susceptible to the virus and supported high levels of thrips reproduction. The TSWV could not be transmitted to Galinsoga parviflora and Sonchus oleraceus by F. occidentalis despite them being highly susceptible in mechanical transmission tests. There was a significant correlation between feeding damage and number of larvae of F. occidentalis on different weeds. Occurrence of weeds that support thrips reproduction and are good hosts of TSWV is a clear indicator of their role in epidemiology and the importance of their management for disease control.     

Selection of tomato plants resistant to a local Polish isolate of tomato spotted wilt virus (TSWV)

We found that the Sw-5 gene confers resistance to one of the Polish isolates of tomato spotted wilt virus (TSWV). A series of tomato breeding accessions was analysed along with standards of resistance and susceptibility to TSWV. The presence of the Sw-5 gene was determined using the available PCR marker. Subsequently plants from these accessions were grown in the presence of the TSWV isolate from Poland. Some of them developed severe symptoms of the TSWV disease. Expression of the virus proteins was also assayed in tissues of the investigated plants. We found general agreement between either lack or presence of the disease symptoms, virus proteins and resistance gene. Some observed discrepancies of these data are also discussed. Our results indicate that marker-assisted selection can be used for breeding of the TSWV-resistant tomato in Poland.     

A multi-generation analysis of the stability of transgenic virus resistance in doubled-haploid tobacco lines

Plants can be genetically engineered for virus resistance by transformation with a viral gene. We transformed tobacco with the tomato spotted wilt virus (TSWV) nucleocapsid gene from the Hawaiian L isolate in order to obtain TSWV resistant breeding lines. Doubled-haploid lines were produced from primary transgenic plants that were selected for resistance to the virus. Several of these lines showed very high levels of resistance and were symptomless after inoculation with the Hawaiian L isolate of TSWV. The accumulation of only low levels of full-length transgene RNA and protein observed in these lines is consistent with an RNA-mediated mechanism of resistance. The lines that were highly resistant to the Hawaiian L isolate of TSWV were also found to be highly resistant to several other isolates of TSWV, while lines that were only moderately resistant to the Hawaiian L isolate were often susceptible to the other isolates. The highly resistant lines were advanced over several generations by self-pollination. Although these lines were fully homozygous, several lines lost resistance in later generations, indicating that the resistance was unstable. Selection for resistance in these unstable lines did not prevent the occurrence of susceptible progeny in subsequent generations. Therefore, testing over several generations is required to determine the stability of resistance when breeding crops with transgenic virus resistance.     

Selection, biological properties and fitness of resistance-breaking strains of Tomato spotted wilt virus in pepper

In glasshouse tests, infective sap from plants infected with 17 different isolates of Tomato spotted wilt virus (TSWV) from four Australian states was inoculated to three Capsicum chinense accessions (PI 152225, PI 159236 and C00943) carrying single genes that confer hypersensitive resistance to TSWV. The normal response to inoculation was development of necrotic (hypersensitive) local lesions in inoculated leaves without systemic invasion, but 3/1386 infected plants also developed systemic susceptible reactions in addition to hypersensitive ones. Similarly when two isolates were inoculated to C. chinense backcross progeny plants, 1/72 developed systemic susceptible reactions in addition to localised hypersensitive ones. Using cultures from the four plants with susceptible reactions and following three to five further cycles of serial subculture in TSWV‐resistant C. chinense plants, four isolates were obtained that gave systemic susceptible type reactions in the three TSWV‐resistant accessions, and in TSWV‐resistant cultivated pepper (C. annuum). When three of these isolates were inoculated to tomato (Lycopersicon esculentum) breeding lines with single gene resistance to TSWV, resistance was not overcome. Similarly, none of the four isolates overcame partial resistance to TSWV in Lactuca virosa. When TSWV isolates were inoculated to tomato breeding lines carrying partial resistance from L. chilense, systemic infection developed which was sometimes followed by ‘recovery’. After four successive cycles of serial passage in susceptible cultivated pepper of a mixed culture of a resistance‐breaking isolate with the non resistance‐breaking isolate from which it came, the resistance‐breaking isolate remained competitive as both were still found. However, when the same resistance‐ breaking isolate was cultured alone, evidence of partial reversion to wild‐type behaviour was eventually obtained after five but not four cycles of long term serial subculture in susceptible pepper, as by then the culture had become a mixture of both types of strain. This work suggests that resistance‐breaking strains of TSWV that overcome single gene hypersensitive resistance in pepper are relatively stable. The findings have important implications for situations where resistant pepper cultivars are deployed widely in the field without taking other control measures as part of an integrated TSWV management strategy.     

Tomato spotted wilt virus (TSWV) in Greece: its incidence following the expansion of Frankliniella occidentalis, and characterisation of isolates collected from various hosts

Leaf samples were collected from plants with tospovirus‐like symptoms from various hosts in different regions of Greece where Thrips tabaci, Frankliniella occidentalis or both vectors occur. The viruses infecting these plants were identified with polyclonal antibodies raised against the N proteins of Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus (INSV) by ELISA. All samples tested positive for TSWV, but not for INSV. ELISA of thirty three isolates, using monoclonal antibodies against the N protein of TSWV, revealed the existence of five epitopes on the N protein. RT‐PCR tests on a few randomly‐selected isolates, using a pair of universal primers, a pair of primers specific for the L gene and a pair of primers specific for the N gene, as well as sequence analysis of a part of the S gene of one isolate, confirmed the authenticity of the virus isolated as TSWV. Host range studies showed differences in susceptibility, especially among species belonging to the Leguminosae and Cucurbitaceae. The species Beloporone guttata and Coleus sp. are reported for the first time as hosts of the virus, whereas Solanum melongena, Celosia cristata, Dianthus chinensis, Stephanotis floribunda and Catharanthus roseus were identified as new hosts of TSWV in Greece.     

Effectiveness of tomato-spotted wilt virus management tactics

Tomato-spotted wilt (TSW) is caused by the tomato-spotted wilt virus (TSWV) and is a major disease affecting the production of tomato and pepper in the Southeastern United States. Before initiating a multistate, regional project addressing this issue, a survey was conducted in North Carolina, South Carolina, Georgia, and Florida to assess the need for improved management of TSWV in these crops. We investigated farmer's stated effectiveness of four common TSWV management tactics (reflective mulch, resistant cultivar, imidacloprid, and Actiguard) in pepper and tomato production using logistic regression. We found that the odds that farmers were satisfied with the use of reflective mulch alone in controlling TSWV is 9-1, about one and a half times the amount obtained from using Actiguard alone. Moreover, the odds that farmers were satisfied with a practice that combines reflective mulch and Actiguard was far greater than that obtained from using each of the technique separately. We found some similarities between farmer's stated effectiveness and revealed effectiveness from experiments.     

Transmission of Tomato spotted wilt virus isolates able and unable to overcome tomato or pepper resistance by its vector Frankliniella occidentalis

Tomato spotted wilt virus (TSWV) causes serious diseases of many economically important crops. Disease control has been achieved by breeding tomato and pepper cultivars with the resistance genes Sw‐5 and Tsw, respectively. However, TSWV isolates overcoming these genetic resistances have appeared in several countries. To evaluate the risk of spread of these resistance‐breaking isolates, we tested their ability of transmission by the main vector of TSWV, the thrips Frankliniella occidentalis. We compared the transmission rate by thrips of six TSWV isolates of different biotype (able or unable to overcome this resistance in pepper and tomato), and with divergent genotype (A and B). Our results indicate that the transmission rate was related to the amount of virus accumulated in thrips but not to virus accumulation in the source plants on which thrips acquired the virus. No correlation was found between transmission efficiency by thrips and the genotype or between transmission efficiency and the ability of overcoming both resistances. This result suggests that resistance‐breaking isolates have the same potential to be transmitted as the isolates unable to infect resistant tomato and pepper cultivars.     

A Survey of Tospoviruses in Bulgaria

A survey of tospoviruses present in Bulgaria was conducted during three vegetation periods. A total of 258 different virus isolates were obtained from tobacco, tomato, weeds and ornamentals in the open fields and from glasshouses. Virus isolates were identified by inoculation to test plants and serology as Tomato spotted wilt virus (TSWV). No indications were obtained that other Tospovirus species were present in Bulgaria. In addition the transmission ability and efficiency of several Bulgarian populations of Thrips tabaci (Lind.) and Frankliniella occidentalis (Perg.) was determined. The experimental results indicated that Bulgarian TSWV isolates are transmitted with high efficiency by F. occidentalis and that only arrhenotokous and not thelytokouse populations of T. tabaci could transmit TSWV.     

Patterns of spread of Tomato spotted wilt virus in field crops of lettuce and pepper: spatial dynamics and validation of control measures

Patterns of spread of Tomato spotted wilt virus (TSWV) were examined in lettuce and pepper plantings into which thrips vectors spread the virus from external virus sources. These plantings were: 1) seven separate field trials into which TSWV ‘infector’ plants of tomato were introduced alongside or near to plantings of lettuce or pepper, and 2) three commercial lettuce plantings into which spread from nearby external infection sources was occurring naturally. The vector thrips species were Frankliniella occidentalis, F. schnitzel and Thrips tabaci, at least two of which were always present. Spatial data for plants with TSWV infection collected at different stages in the growing period were assessed by plotting gradients of infection, and using Spatial Analysis by Distance IndicEs (SADIE) and maps of spatial pattern. Despite the persistent nature of TSWV transmission by thrips vectors, in both lettuce and pepper plantings there was a steep decline in TSWV incidence with distance from external infection sources that were alongside them. The extent of clustering increased over time and was greatest closest to the source. The relationship between percentage infection and assessment date suggested that spread was predominantly monocyclic with only limited polycyclic spread. Development of isolated clusters of infected plants distant from TSWV sources within both crops was consistent with only limited polycyclic spread. Spread to lettuce was greater downwind than upwind of virus source, with magnitude and proximity of source determining the amount of spread. When 15 m wide fallow or non-host (cabbage) barriers separated TSWV sources from lettuce plantings, spread was slower and there was much less clustering with the latter. In commercial lettuce plantings, spread was favoured by TSWV movement within successive side-by-side plantings. The spatial data from the diverse scenarios examined enabled recommendations to be made over ‘safe’ planting distances between external infection sources of different magnitudes and susceptible crops that were short-lived (e.g. lettuce) or long-lived (e.g. pepper). They also helped validate the inclusion of isolation and ‘safe’ planting distances, planting upwind, prompt removal of virus sources, avoidance of side-by-side plantings, and deploying intervening non-host barrier crops as control measures within an integrated disease management strategy for TSWV in field vegetable crops.     

Tactics for management of thrips (Thysanoptera: Thripidae) and tomato spotted wilt virus in tomato

Four studies were conducted in Georgia during spring 1999, 2000, 2001, and 2002 to evaluate various management tactics for reducing thrips and thrips-vectored tomato spotted wilt virus (TSWV) in tomato and their interactions relative to fruit yield. Populations of thrips vectors of TSWV, Frankliniella occidentalis (Pergande) and Frankliniella fusca (Hinds), were determined using flower and sticky trap samples. The management practices evaluated were host plant resistance, insecticide treatments, and silver or metallic reflective mulch. Averaged over all tests, the TSWV-resistant tomato 'BHN444' on silver mulch treatment had the largest effect in terms of reducing thrips and spotted wilt and increasing marketable yield. Of the insecticide treatments tested, the imidacloprid soil treatment followed by early applications of a thrips-effective foliar insecticide treatment provided significant increase in yield over other treatments. Tomato yield was negatively correlated with the number of F. fusca and percentage of TSWV incidence. F. occidentalis per blossom was positively correlated with percentage of TSWV incidence, but not with yield. No significant interactions were observed between cultivar reflective mulch main plot treatments and insecticide subplot treatments; thus, treatment seemed to be additive in reducing the economic impact of thrips-vectored TSWV. Control tactics that manage thrips early in the growing season significantly increased tomato yield in years when the incidence of TSWV was high (>17%).     

Seasonal dispersal patterns of Frankliniella fusca (Thysanoptera: Thripidae) and tomato spotted wilt virus occurrence in central and eastern North Carolina

The seasonal abundance and temporal pattern of Frankliniella fusca Hinds dispersal were monitored from 1996 to 2000 at 12 locations in central and eastern North Carolina. The predominant vector species of tomato spotted wilt virus (TSWV) captured across all locations was F. fusca (98%). The temporal patterns of F. fusca dispersal observed during spring seasons varied among locations in all years except 2000. Regression analysis estimated that times of first flight in the spring seasons varied among locations, whereas flight duration intervals were similar. Temporal patterns of F. fusca captured varied significantly between aerial traps placed 0.1 and 1.0 m above the soil surface. Fewer total thrips were captured at 0.1 m, although thrips dispersal occurred earlier and over a greater time interval compared with 1.0-m traps. Temporal patterns of TSWV occurrence differed among locations in the spring seasons of 1999 and 2000, whereas patterns of virus occurrence were similar during the fall seasons. Patterns of F. filsca dispersal and subsequent TSWV occurrence were synchronous at locations in 1999 and 2000 where the greatest number of TSWV lesions was recorded. Knowledge of the temporal patterns of F. fiasca dispersal and TSWV occurrence may be a useful indicator for describing the time when susceptible crops are at highest risk of TSWV infection.     

Mitigation of Tomato Spotted Wilt Virus Infection and Symptom Expression by Water Stress

The effects of water stress on the infection and systemic movement of tomato spotted wilt virus (TSWV) in vegetative tomato plants are examined. Two groups of plants: water stressed (water potential ?1.31 MPa) and well-watered ones (water potential ?0.65 MPa), were mechanically inoculated on the third expanded leaf from the top. Each group was then divided into a well-watered and a water stressed one. TSWV inoculation was effective both on water-stressed and well-watered plants. In all groups, infection was first detected in roots, then in shoot apices and subsequently in the rest of the leaves. Systemic virus infection was somewhat slower in the water stressed plants. Post inoculation stress treatments had a significant effect in reducing infection as well as in attenuating TSWV symptoms. Results from the present work suggest that water stress may be effective in controlling virus concentration and systemic infection symptoms in tomato plants and could thus be used as a tool to study the regulation of development of TSWV infection in this system.     

A novel tomato spotted wilt virus isolate encoding a noncanonical NSm C118F substitution associated with Sw-5 tomato gene resistance breaking

The nonstructural protein NSm of tomato spotted wilt virus (TSWV) has been identified as the avirulence determinant of the tomato single dominant Sw-5 resistance gene. Although Sw-5 effectiveness has been shown for most TSWV isolates, the emergence of resistance-breaking (RB) isolates has been observed. It is strongly associated with two point mutations (C118Y or T120N) in the NSm viral protein. TSWV-like symptoms were observed in tomato crop cultivars (+Sw-5) in the Baja California peninsula, Mexico, and molecular methods confirmed the presence of TSWV. Sequence analysis of the NSm 118–120 motif and three-dimensional protein modelling exhibited a noncanonical C118F substitution in seven isolates, suggesting that this substitution could emulate the C118Y-related RB phenotype. Furthermore, phylogenetic and molecular analysis of the full-length genome (TSWV-MX) revealed its reassortment-related evolution and confirmed that putative RB-related features are restricted to the NSm protein. Biological and mutational NSm 118 residue assays in tomato (+Sw-5) confirmed the RB nature of TSWV-MX isolate, and the F118 residue plays a critical role in the RB phenotype. The discovery of a novel TSWV-RB Mexican isolate with the presence of C118F substitution highlights a not previously described viral adaptation in the genus Orthotospovirus, and hence, the necessity of further crop monitoring to alert the establishment of novel RB isolates in cultivated tomatoes.     

Relationships between Tospovirus Incidence and Thrips Populations on Tomato in Mendoza, Argentina

The objectives of this work were to estimate the capability of local populations of thrips as vectors of groundnut ringspot virus (GRSV) and tomato spotted wilt virus (TSWV), and to determine the species composition of vectors in tomato crops. Transmission assays were performed. Incidence of tospoviruses was estimated in commercial crops. Random samples of flowers were taken from tomato for identification of thrips. Of the five species of thrips tested, Frankliniella gemina (first record), F. occidentalis and F. schultzei transmitted GRSV and TSWV. F. schultzei was a significantly more efficient vector of GRSV than F. occidentalis under controlled assay conditions. The thrips were identified on flowers from six surveyed tomato crops. F. occidentalis was the most frequently identified species (43.0%), followed by F. schultzei (35.6%) and Thrips tabaci (10.1%). The incidence of tospoviruses was low (1.1–2.8%) in crops planted during August–September and greater (9.5–61.1%) in crops planted in December. GRSV was prevalent (85%) over TSWV (11%).     

Variation in thrips species composition in field crops and implications for tomato spotted wilt epidemiology in North Carolina

Thrips were surveyed in tomato spotted wilt-susceptible crops in five areas across North Carolina. Tomato, pepper, and tobacco plants in commercial fields were sampled and 30 species of thrips were collected over a 3-year period. The most common species overall was Frankliniella tritici (Fitch). The most common thrips species that are known to vector Tomato Spotted Wilt Virus (TSWV) were F. fusca (Hinds), and F. occidentalis (Pergande). Relatively low numbers of Thrips tabaci Lindeman, another reported vector, were collected. The spatial and temporal occurrence of vectors varied with sampling method, crop species, region of North Carolina, and localized areas within each region. In a laboratory experiment, no difference was detected between the ability of F. fusca and F. occidentalis to acquire and transmit a local isolate of TSWV. Based on vector efficiency and occurrence, F. fusca is considered the most important vector of TSWV in tobacco, whereas both F. fusca and F. occidentalis are important vectors of TSWV in tomato and pepper.     

Phylogenetic analysis based on full genome sequencing of Italian tomato spotted wilt virus isolates identified in “Roggianese” sweet pepper and chilli pepper

A study aimed at defining population structure of Italian tomato spotted wilt virus (TSWV) isolates was performed. Full genome sequencing of six TSWV isolates found in two Italian regions (two from Latium: Lazio 17 and Tarquinia; and four from Calabria: PepCal 10, 12, 22 and 24) were assembled. Identity percentages in nucleotide sequence among these TSWV isolates are here provided. The six full length genome sequences were compared with other two Italian isolates (p105 and p202/3WT) already fully sequenced, as well as full TSWV genomes that could be retrieved from GenBank. Phylogenetic analysis, performed in concatenated sequences and for each gene of each genome segment (L, M and S), confirmed the presence of two clades, namely A-like and D-like. In particular, the phylogenetic tree based on segment L grouped all the newly sequenced TSWV isolates in D-like clade. In the M segment phylogenetic tree, all our TSWV isolates shifted in the A-like clade. Isolates separation was not correlated to their geographical origin in phylogenetic study of distinct ORFs encoded by the RNA S segment. In fact, in nucleocapsid-encoding phylogenetic tree, PepCal 10 and 22 grouped in an A-like clade with p105, PepCal 12 and 24 in a D-like clade with p202/3WT, whereas Lazio 17 and Tarquinia in a third well distinct group. NSs tree displayed only PepCal 10 with p105 in A-like clade, whereas PepCal 12, 22, 24 with p202/3WT in D-like subclade; and isolates from Latium grouped a separated clade adjacent to D-like isolates. Additional analysis on putative reassortment events showed that TSWV Calabrian isolates likely originated from a reassortment event in M RNA and other in S RNA with p105 as major parent. Recombination events were detected in isolates from Latium in L and S RNAs with Chinese isolates as putative major parent.     

Selection of resistance breaking strains of tomato spotted wilt tospovirus

In glasshouse tests, sap from plants infected with 15 different isolates of tomato spotted wilt tospovirus (TSWV) from three Australian states was inoculated to nine genotypes of tomato carrying TSWV resistance gene Sw-5 or one of its alleles. A further two resistant tomato genotypes were inoculated with four isolates each. The normal response in resistant genotypes was development of necrotic local lesions in inoculated leaves without systemic invasion, but 22/752 plants also developed systemic reactions in addition to local hypersensitive ones. Using cultures from two of these systemically infected plants and following four cycles of subculture in TSWV resistant tomato plants, two isolates were obtained that gave susceptible type systemic reactions but no necrotic spots in inoculated leaves of resistant tomatoes. When these two isolates, Da_WA-1d and To_TAS-1d, were maintained by repeated subculture for 10 successive cycles in Nicotiana glutinosa or a susceptible tomato genotype, they still induced susceptible type systemic reactions when inoculated to resistant tomato plants. They were therefore stable resistance breaking isolates as regards overcoming gene Sw-5. When resistance-breaking isolate Da_WA^-1ld multiplied together with original isolate Da_WA-l in susceptible tomato, it was fully competitive with the original isolate. However, when Da_WA-ld and To_TAS-ld were inoculated to TSWV resistant Lycopersicon peruvianum lines PI 128660R and PI 128660S and to TSWV resistant Capsicum chinense lines PI 152225, PI 159236 and AVRDC CO0943, they failed to overcome the resistance, producing only necrotic local lesions without systemic infection. Thus, although the ease of selection, stability and competitive ability of resistance breaking isolates of TSWV is cause for concern, L. peruvianum and C. chinense lines are available which are effective against them. The effectiveness of the resistance to TSWV in nine tomato genotypes was examined in a field experiment. Spread was substantial in the susceptible control genotype infecting 42% of plants. Resistance was ineffective in cv. Bronze Rebel, 26% of plants developing infection. In contrast, it held up well in the other eight resistant genotypes with only 1–3 or no plants of each becoming infected. Accumulated numbers of Thrips tabaci, Frankliniella occidentalis and F. schultzei were closely correlated with TSWV spread.