西花蓟马传播番茄斑萎病毒研究进展

西花蓟马Frankliniella occidentalis是世界性重要检疫性害虫之一,不仅直接取食危害作物而且传播病毒,从而造成极为严重的经济损失。由于西花蓟马在我国具有广泛的适生范围,随其入侵我国并随之传播的番茄斑萎病毒(Tomato spotted wilt virus)已在我国不同地域发现,对经济作物已形成严重威胁。本文综述了西花蓟马对番茄斑萎病毒的获取、携带和传播扩散过程及其病毒在蓟马体内的循环过程和机理,总结了影响西花蓟马传播番茄斑萎病毒效率的因素,并评述了西花蓟马-病毒-植物这一互作系统及其对西花蓟马生长发育适合度的影响,以期为我国西花蓟马传播番茄斑萎病毒的基础研究和防控提供理论依据与指导。     

Characterization of a vector-non-transmissible isolate of Tomato spotted wilt virus

The characteristics of a thrips‐non‐transmissible isolate of Tomato spotted wilt virus (TSWV), designated TSWV‐M, were compared with those of a thrips‐transmissible isolate, designated TSWV‐T. TSWV‐M showed a narrower host range than TSWV‐T. Adult thrips failed to transmit TSWV‐M, although the vector acquired the virus during the larval stages. TSWV‐M was detected by RT‐PCR in adult thrips bodies, but not in thrips heads, suggesting that loss of thrips transmissibility was the result of the absence of virus in adult thrips salivary glands. Whereas N (nucleoprotein), NSs (non‐structural protein) and G_C (the C‐terminal portion of the glycoprotein precursor protein) were present in similar amounts in leaf tissue from TSWV‐M‐ or TSWV‐T‐infected plants, G_N (the N‐terminal portion of the glycoprotein precursor protein) was present at much lower amounts in TSWV‐M‐ than in TSWV‐T‐infected plants. SDS‐PAGE and immunoblotting analysis of TSWV‐M and TSWV‐T virion preparations with G_N‐ and G_C‐specific antibodies revealed similar amounts of the G_N and G_C glycoproteins in TSWV‐T virions, but lower amounts of G_N than G_C in TSWV‐M virions. This resulted in a statistically significant reduction in the G_N/G_C ratio in TSWV‐M virions. In affinoblots, the G_C and G_N glycoproteins of TSWV‐M exhibited weak binding with lectins showing affinity for N‐linked oligosaccharide structures. Sequence analysis of M RNA (medium segment of the TSMV genome) revealed no deletions or frameshift mutations in the G_N/G_C precursor of TSWV‐M. However, five amino acid changes were detected in the G_N/G_C precursor. A single, relatively conservative amino acid substitution (V→I) was observed in the NSm protein. Sequence analysis of S RNA (small portion of the TSMV genome) revealed a large intergenic region with no changes in the N protein and with three amino acid changes in the NSs protein.     

Ornamental plants and thrips populations associated with tomato spotted wilt virus in Greece

A survey was conducted in order to record the ornamental plants that are hosts of tomato spotted wilt virus (TSWV) and impatiens necrotic spot virus (INSV) in Greece. Polyclonal antibodies prepared against the N protein of a Greek isolate of TSWV fromGerbera jamesonii (GR-34) were used. Leaf samples were taken from plants showing typical symptoms of tospovirus infection such as chlorotic and necrotic rings on the leaves and malformation and necrosis of the flowers. The samples were tested by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using polyclonal antibodies to the N proteins of TSWV and INSV (NL-07). ELIS A-positive samples were mechanically transmitted to plants ofPetunia hybrida, Nicotiana rustica andN. benthamiana to confirm infection. Although none of the samples was found infected with INSV, TSWV presence was recorded in 42 botanical species that belong to 40 genera in 27 families. Among them the speciesBeloperone guttata, Coleus barbatus, Impatiens petersiana andLilium auratum are reported for the first time as hosts of TSWV, whereasBegonia sp.,Catharanthus roseus Celosia cristata, Dianthus chinensis, Fuchsia hybrida andStephanotis floribunda are found as new hosts of the virus in Greece. Thrips collected from TSWV-infected plants were in most cases identified asFrankliniella occidentalis, except from plants ofDendranthema sp. andDianthus caryophyllus whereThrips tabaci individuals were also identified. Different percentages of transmitters were noticed when the thrips populations collected from TSWV-infected ornamental hosts were tested for transmission of TSWV.     

北京辣椒和番茄上番茄斑萎病毒的鉴定

北京大兴区某蔬菜种植基地的辣椒和番茄发现疑似病毒感染,造成较大经济损失.经症状分析和RT-PCR检测,证实其感染番茄斑萎病毒(tomato spotted wilt virus,TSWV).基于TSWV序列特征对该地区TSWV进化及来源进行分析,结果显示其与北京顺义区已经报道TSWV毒株进化关系较近,但较国内其他地区株...     

Purification and serological analysis of tomato spotted wilt virus

A modified procedure for the purification of TSWV is described which consists of differential centrifugation, treatment with antiserum against sap from healthyN. rustica and density-gradient centrifugation in sucrose gradients. Centrifugation in the zonal-rotor proved efficient for processing large amounts of infected leaves.Antisera to purified TSWV were produced in rabbits. These sera reacted with healthyN. rustica sap. This reaction could be abolished by absorption with either healthyN. rustica sap or polysaccharides purified fromN. rustica. Unabsorbed sera reacted with the major TSWV structural proteins following their electrophoretic separation in polyacrylamide-gels. The positions of the precipitin lines corresponded with those of the structural proteins visualized by staining. The intensity of these lines was greatly diminished after absorption of the antisera with either healthyN. rustica sap or polysaccharides. It was concluded that the reaction of TSWV antiserum with the structural proteins is partly due to host-derived polysaccharides covalently bound to the virus proteins.Samenvatting Het bronsvlekkenvirus van de tomaat (TSWV) werd gezuiverd met een procedure die bestond uit een cyclus van differentiële centrifugering, behandeling van de virus-suspensie met antiserum tegen bestanddelen van gezonde planten (N. rustica) en dichtheidsgradiëntcentrifugering (Fig. 1 en Fig. 3). De zone rotor kon worden gebruikt wanneer grote hoeveelheden bladmateriaal werden gebruikt (Fig. 2).Twee antisera, die elk een verschillende activiteit tegen TSWV (Tabel 1, Fig. 4) hadden, werden bereid. Deze sera reageerden ook met sap en polysacchariden uitN. rustica (Fig. 6). Door verzadiging met polysacchariden werd de reactie tegen TSWV aanmerkelijk verzwakt. Analyse van de eiwitbandjes die na polyacrylamidegel-elektroforese waren gevormd, toonde aan dat de activiteit van de sera tegen de eiwitcomponenten van TSWV was gericht (Fig. 5) en dat deze activiteit na verzadiging met polysacchariden zwakker was. Uit deze reacties kan geconcludeerd worden dat polysacchariden van de waardplant aan de structurele eiwitten van TSWV zijn gebonden. Niet geabsorbeerd antiserum geeft met TSWV drie precipitatielijnen (Fig. 4), maar na absorptie met compleet virus trad nog slechts één precipitatielijntje op (Fig. 7). Mogelijk wordt dit lijntje gevormd door het nucleocapside-eiwit dat door de dissociatie van het virus vrij komt. Sera die tegen normale bestanddelen vanN. rustica zijn bereid, vormen geen zichtbare reactie met compleet virus; wel wordt er een lijntje gevormd indien het virus met guanidine-HCl wordt gedissociëerd (Fig. 8). Deze waarneming kan verklaren dat deze antisera kunnen worden gebruikt bij de zuivering van TSWV ondanks het feit dat dit virus polysacchariden uit de waardplant draagt.     

由美国进境生菜种子上番茄斑萎病毒的检测鉴定

采用DAS-ELISA检测方法,从美国进境的生菜种子中筛选出携带有番茄斑萎病毒(Tomato spotted wilt virus,TSWV)的疑似阳性样品,通过建立的RT-PCR方法对酶联免疫吸附测定阳性样品进行复核鉴定,并进一步测序。测序结果与GenBank中已有的番茄斑萎病毒分离物KT160282、AY744474、GU369726、KP330470的序列同源性达到96%。结果表明,TSWV可通过种子携带入境。     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001     

Assessing the susceptibility of chrysanthemum cultivars to tomato spotted wilt virus

Three methods were compared to assess the susceptibility of vegetatively propagated chrysanthemum to tomato spotted wilt tospovirus (TSWV): mechanical and thrips-mediated inoculation of whole plants, and a leaf-disc assay. As symptom expression was often poor or even absent, TSWV infections and subsequent susceptibility to TSWV were determined by ELISA. All 15 chrysanthemum cultivars tested were susceptible to TSWV, irrespective of their degree of vector resistance (based on feeding-scar damage rates). Thrips-mediated inoculation using different numbers of thrips revealed that 100% infection was obtained when plants were challenged by six thrips per plant, whereas 80 and over 50%, respectively, of the plants became infected when inoculated by a single male or female thrips. However, false negatives were scored even after intensive sampling because of erratic, cultivar-specific and time-dependent virus distribution after inoculation in the plants. Labour-intensive samplings and long incubation periods could be overcome by a readily applicable leaf-disc assay. This assay was as reliable as thrips-mediated inoculation of whole plants, and its use is therefore favoured to assess chrysanthemum cultivars for TSWV susceptibility.     

Inheritance of resistance to acrinathrin in Frankliniella occidentalis (Thysanoptera: Thripidae)

BACKGROUND: The western flower thrips (WFT), Frankliniella occidentalis (Pergande), is an economically important pest. The genetic basis of acrinathrin resistance was investigated in WFT. RESULTS: The resistant strain, selected in the laboratory for acrinathrin resistance from a pool of thrips populations collected in Almeria (south-eastern Spain), showed a high resistance to acrinathrin (43-fold based on LC(50) values) compared with the laboratory susceptible strain. Mortality data from reciprocal crosses of resistant and susceptible thrips indicated that resistance was autosomal and not influenced by maternal effects. Analysis of probit lines from the parental strains and reciprocal crosses showed that resistance was expressed as a codominant trait. To determine the number of genes involved, a direct test of monogenic inheritance based on the backcrosses suggested that resistance to acrinathrin was probably controlled by one locus. Another approach, which was based on phenotypic variances, showed n(E), or the minimum number of freely segregating genetic factors for the resistant strain, to be 0.79. CONCLUSION: The results showed that acrinathrin resistance in WFT was autosomal and not influenced by maternal effects, and was expressed as a codominant trait, probably controlled by one locus.     

Detection of tomato spotted wilt virus and transmission by Frankliniella occidentalis in France

Tomato spotted wilt virus (TSWV) was isolated from pepper, tomato, eggplant, broad bean, lettuce, basil, chrysanthemum, aster, New Guinea impatiens, anemone and gloxinia plants. Virus identification was based on host range, vector transmission, serology and electron microscopy. TSWV was readily detected by ELISA in naturally or artificially infected cultivated or weed plants. The virus was also detected in individual F. occidentalis thrips. The spread of TSWV in vegetable and ornamental plants in greenhouses and/or in the open is related to the close relationship of the virus with the vector.     

Occurrence and characterization of tomato spotted wilt orthotospovirus isolated from cucumber

European Journal of Plant Pathology - Diseases associated with tomato spotted wilt orthotospovirus (TSWV) are a serious threat to vegetable production worldwide. In 2019, leaf necrosis disease was...     

Biological and molecular characterization of tomato spotted wilt virus (TSWV) resistance-breaking isolates from Argentina

Tomato spotted wilt virus (TSWV) has been present in Argentina since 1938 and had limited sweet pepper and tomato production until the introduction of resistant cultivars bearing Tsw and Sw-5b genes. However, the wide use of TSWV-resistant pepper plants in La Plata Horticultural Belt (LPHB) triggered the emergence of resistance-breaking isolates (RB), increasing the economic impact of TSWV in pepper. This work characterized 11 natural RB pepper isolates from LPHB that have overcome the Tsw resistance gene in Capsicum sp. but are unable to break the Sw-5b-mediated resistance in tomato. Phylogenetic analysis of the N gene showed that the LPHB isolates are most closely related to isolates from Asia, indicating that Argentine TSWV isolates might have emerged from the Asian continent. The NSs sequence analysis reinforces the hypothesis that the appearance of an RB phenotype is a consequence of a number of different single amino acid substitutions spread along the NSs gene that lead to multiple independent evolutionary events. These results provide information on the current situation of the tospovirus–pepper/tomato pathosystems in LPHB, which represents a fundamental prerequisite to include these RB isolates in future screening programmes in order to select new and durable sources of resistance to TSWV in pepper.     

An electron microscope study of tomato spotted wilt virus in the plant cell

Characteristic particles of about 70 m diameter were found in tomato spotted wilt virus (TSWV) infected cells of roots, stems, leaves and petals of several plant species. These particles were similar in size and form to those found in the pellets of partially purified infective TSWV.     

Response of Cucurbita spp. to tomato leaf curl New Delhi virus inoculation and identification of a dominant source of resistance in Cucurbita moschata

Tomato leaf curl New Delhi virus is an emerging whitefly-borne species of begomovirus in Mediterranean regions that poses a severe threat to cucurbit crops of the genus Cucurbita. Until now, only two sources of resistance have been identified in Cucurbita spp., these being PI604506 (cv. Large Cheese) and PI381814 (Indian landrace), both of C. moschata. The resistance of cv. Large Cheese is conferred by a single recessive gene located on chromosome 8. The objective of the present investigation was to screen for tomato leaf curl New Delhi virus (ToLCNDV) resistance among 105 accessions drawn from five species of Cucurbita and, if high resistance was found in any of them, determine the mode of inheritance. Screening was conducted using whitefly-mediated inoculation on all 105 accessions. The accessions showing some resistance were further screened by mechanical inoculation as well as by quantitative PCR-based diagnostics. The results showed that, overall, the accessions of C. pepo and C. maxima were the most susceptible, those of C. argyrosperma and C. ecuadorensis intermediate, and those of C. moschata most resistant to ToLCNDV. Only one accession of C. moschata, BSUAL-252, originating from Japan, was highly resistant to ToLCNDV, showing no symptoms after either method of inoculation, and absence of virus accumulation. Upon crossing BSUAL-252 with a susceptible accession of C. moschata, BSUAL-265, the resistance was observed to be conferred by a single dominant gene. This gene is not linked to the genomic region on chromosome 8 where the locus of the previously identified recessive gene for ToLCNDV resistance resides.     

Transmission of tomato spotted wilt tospovirus by Thripstabaci populations originating from leek

The transmission of tomato spotted wilt tospovirus (TSWV) by Thrips tabaci collected from leek was studied using the petunia local-lesion leaf-disc assay. After an acquisition-access period of 72 h given to newborn larvae up to 8 h old, the efficiency of transmission by adults was determined in three inoculation-access periods of 48 h. This efficiency varied for six T. tabaci populations from 0.7 to 11.6% in experiments using the Greek TSWV isolate GR-04. Males were more efficient transmitters than females (19 out of 176 versus five out of 494). Frankliniella occidentalis transmitted the same virus with a higher efficiency (34.8%). The transmission rate differed also among TSWV isolates, as shown in tests with four T. tabaci using two isolates. The virus was more efficiently acquired from infected leaf material of Datura stramonium than from that of Emilia sonchifolia . Plants of the latter species were more susceptible than Nicotiana tabacum in thrips transmission tests.     

Development of an assay system using thrips-mediated inoculation to evaluate resistance of Capsicum spp. to Tomato spotted wilt virus

Considerable losses in pepper production by Tomato spotted wilt virus (TSWV) have been reported worldwide. In breeding programs, an assay for resistance that accurately estimates field occurrence of TSWV during pepper production is critical because the virus is vector transmitted. Here, we establish an assay system of TSWV-resistant Capsicum spp. using insect-mediated inoculation within an acrylic chamber in which environmental conditions such as temperature, light intensity, and nutrient supply are controlled. This chamber enables transmission of TSWV from viruliferous plants to plants used in the resistance assay with Frankliniella occidentalis safely, quickly, and precisely.