Complexity and Local Specificity of the Virome Associated with Tospovirus-Transmitting Thrips Species.

Frankliniella occidentalis (western flower thrips [WFT]) and Thrips tabaci (onion thrips [OT]) are insect species that greatly impact horticultural crops through direct damage and their efficient vectoring of tomato spotted wilt virus and iris yellow spot virus. In this study, we collected thrips of these species from 12 field populations in various regions in Italy. We also included one field population of Neohydatothrips variabilis (soybean thrips [ST]) from the United States. Total RNA data from high-throughput sequencing (HTS) were used to assemble the virome, and then we assigned putative viral contigs to each thrips sample by real-time reverse transcription-quantitative PCR (qRT-PCR). Excluding plant and fungal viruses, we were able to identify 61 viral segments, corresponding to 41 viruses: 14 were assigned to WFT, 17 to OT, and 1 to ST; 9 viruses could not be assigned to any species based on our stringent criteria. All these viruses are putative representative of new species (with only the exception of a sobemo-like virus that is 100% identical to a virus recently characterized in ST) and some belong to new higher-ranking taxa. These additions to the viral phylogeny suggest previously undescribed evolutionary niches. Most of Baltimore's classes of RNA viruses were present (positive- and minus-strand and double-stranded RNA viruses), but only one DNA virus was identified in our collection. Repeated sampling in a subset of locations in 2019 and 2020 and further virus characterization in a subset of four thrips populations maintained in the laboratory allowed us to provide evidence of a locally persistent thrips core virome that characterizes each population. IMPORTANCE Harnessing the insect microbiome can result in new approaches to contain their populations or the damage they cause vectoring viruses of medical, veterinary, or agricultural importance. Persistent insect viruses are a neglected component of their microbiota. In this study, for the first time, we characterize the virome associated with the two model systems for tospovirus-transmitting thrips species, of utmost importance for the direct and indirect damage they cause to a number of different crops. The thrips virome characterized includes several novel viruses, which in some cases reveal previously undescribed clades. More importantly, some of the viruses we describe are part of a core virome that is specific and consistently present in distinct geographical locations monitored over the years, hinting at a possible mutualistic symbiotic relationship with their host.

The potential of host plants for biological control of Tuta absoluta by the predator Dicyphus errans.

Dicyphus errans (Wolff) has been shown to be a suitable biocontrol agent for Tuta absoluta (Meyrick). This generalist predator shares various host plants with the exotic pest, and these interactions could be exploited to enhance pest control. Therefore, host preference, survival rate and development times of the predator and prey were investigated on crop and non-crop plant species. Among the tested plants, the favourite hosts were Solanum species for T. absoluta, and herb Robert, European black nightshade, courgette and tomato for D. errans. Tuta absoluta accepted the same plant species as hosts for oviposition, but it never developed on herb Robert and courgette in all the experiments. Based on our results, we would suggest the use of courgette and herb Robert in consociation with tomato and as a companion plant, respectively, which may keep pest densities below the economic threshold. Moreover, the omnivorous and widespread D. errans could be a key predator of this exotic pest, allowing a high encounter probability on several cultivated and non-cultivated plant species.

The NSs protein of tomato spotted wilt virus is required for persistent infection and transmission by Frankliniella occidentalis.

UNLABELLED: Tomato spotted wilt virus (TSWV) is the type member of tospoviruses (genus Tospovirus), plant-infecting viruses that cause severe damage to ornamental and vegetable crops. Tospoviruses are transmitted by thrips in the circulative propagative mode. We generated a collection of NSs-defective TSWV isolates and showed that TSWV coding for truncated NSs protein could not be transmitted by Frankliniella occidentalis. Quantitative reverse transcription (RT)-PCR and immunostaining of individual insects detected the mutant virus in second-instar larvae and adult insects, demonstrating that insects could acquire and accumulate the NSs-defective virus. Nevertheless, adults carried a significantly lower viral load, resulting in the absence of transmission. Genome sequencing and analyses of reassortant isolates showed genetic evidence of the association between the loss of competence in transmission and the mutation in the NSs coding sequence. Our findings offer new insight into the TSWV-thrips interaction and Tospovirus pathogenesis and highlight, for the first time in the Bunyaviridae family, a major role for the S segment, and specifically for the NSs protein, in virulence and efficient infection in insect vector individuals. IMPORTANCE: Our work is the first to show a role for the NSs protein in virus accumulation in the insect vector in the Bunyaviridae family: demonstration was obtained for the system TSWV-F. occidentalis, arguably one of the most damaging combination for vegetable crops. Genetic evidence of the involvement of the NSs protein in vector transmission was provided with multiple approaches.

A member of a new Tospovirus species isolated in Italy from wild buckwheat (Polygonum convolvulus).

Electron microscopy of extracts from diseased Polygonum convolvulus plants from Piedmont (Italy) revealed particles with the morphological features of a tospovirus. Sequencing of the full-length small (S) and medium (M) genome segments indicated that the virus is a member of a new Tospovirus species provisionally named Polygonum ringspot virus. A feature distinguishing it from members of other Tospovirus species was the presence of a very short intergenic region on the S segment lacking the potential for formation of the predicted hairpin structure involved in subgenomic expression. Antibodies made against purified nucleocapsids allowed serological comparison with other tospovirus isolates and revealed a relationship with tomato yellow ring virus, and to a lesser extent, to iris yellow spot virus. Serological tests detected the virus in various locations in northern and central Italy. The experimental host range was wide, although in nature the virus appeared restricted to two Polygonum species.

Two Field Isolates of Tomato Spotted Wilt Tospovirus Overcome the Hypersensitive Response of a Pepper (Capsicum annuum) Hybrid with Resistance Introgressed from C. chinense PI152225.

The hypersensitive response to tomato spotted wilt tospovirus (TSWV) present in Capsicum chinense PI152225 (1) was introgressed into C. annuum cultivars. During the summer of 1998, a hybrid with good agronomic performance was grown in glasshouses in Albenga, Liguzia Region of northwestern Italy, an area where infection by TSWV in pepper has been severe since 1992. In August, observations of different susceptible cultivars revealed that >50% of plants had TSWV-like symptoms, whereas the resistant hybrid remained healthy, except for two plants that showed virus-like symptoms on apical leaves and fruits. From the infected plants, tospoviruses (coded P164/6 and P166) were transmitted by sap-inoculation to Nicotiana benthamiana. Triple-antibody sandwich enzyme-linked immunosorbent assay with a panel of monoclonal antibodies against the TSWV nucleocapsid, but with different reactivity to the related species groundnut ringspot (GRSV) and tomato chlorotic spot (TCSV) viruses, indicated the isolates were TSWV. The host ranges of the isolates were wide and typical of normal TSWV isolates. Thus, they incited typical symptoms in all 50 TSWV-susceptible C. annuum cv. Quadrato d'Asti plants. However, isolate P164/6 also systemically infected 12 of 27 C. chinense PI152225 and 14 of 19 C. chinense PI159236 plants. These accessions are normally resistant to TSWV (1). Isolate P166 systemically infected 7 of 17 C. chinense PI152225 and 6 of 11 C. chinense PI159236 plants. Systemically infected plants showed severe necrosis, and some plants died. Other plants showed only necrotic local lesions. The response by C. chinense differed from that caused by typical TSWV, which causes only local lesions, and from both GRSV and TCSV, which cause mosaic but no necrosis in 100% of plants. The two new TSWV isolates were tested for transmission using a local population of Frankliniella occidentalis in a leaf disk assay with susceptible C. annuum. Transmission rates were high: 93.7% (63 thrips) for isolate P164/6 and 89.9% (49 thrips) for P166. Thus, the fitness of the two TSWV resistance-breaking isolates (a wide experimental host range and high transmission rates by the natural vector) was as high as that of typical TSWV. The absence of systemic infection in some C. chinense PI152225 and PI159236 plants that are resistant to typical TSWV suggests the possibility of selecting plants resistant to these pathotypes. This is the first report of field tospovirus isolates typed as TSWV (according to the current taxonomy based on nucleocapsid serology) overcoming the hypersensitive response of C. chinense PI152225 and PI159236, an ability previously found only in closely related viruses: TCSV and GRSV (2). Other TSWV-like isolates systemic on C. chinense were not typed further (3,4). References: (1) L. L. Black et al. Plant Dis. 75:863, 1991. (2) L. S. Boiteux and A. C. DeAvila. Euphytica 75:139, 1994. (3) H. A. Hobbs et al. Plant Dis. 78:1220, 1994. (4) B. Moury et al. Euphytica 94:45, 1997.