Transmissibility of four tospoviruses by a thelytokous population ofThrips tabaci from Liguria, Northwestern Italy

Studies were carried out on a population ofThrips tabaci Lindeman (Thysanoptera: Thripi-dae) from Liguria to assess its sex-ratio and its ability to transmit four tospoviruses: tomato spotted wilt (TSWV), impatiens necrotic spot, tomato chlorotic spot and groundnut ringspot. The population was composed of females only (therefore thelytokous). The first instar larvae were allowed to acquire the virus for 48 h on infected leaves of datura, basil or pepper, and then reared on cucumber until emergence, which medially occurred 9.5 days after hatching. Transmission capacity was checked using two inoculation access periods (lAPs) of 48 h each on pepper leaf disks.T. tabaci was able to transmit TSWV isolate P105 with an efficiency of 16.7% and 4.4% in the first and second IAP, respectively, and TSWV isolate BR-01 with an efficiency of 2.0%. The onion thrips did not transmit the three other tospoviruses. During the IAPs, almost all adults fed on the leaf disks, producing evident silvery scars. The presence of tospovirus nucleocapsids in thrips was assayed by Triple Antibody Sandwich (TAS) and cocktail ELISA. Not all adults that had transmitted TSWV were positive in the tests, whereas some non-transmitter individuals proved positive. For each of the other tospoviruses, some thrips were positive in at least one test, although none was able to transmit the virus.     

Plant Viruses Transmitted by Whiteflies

One-hundred and fourteen virus species are transmitted by whiteflies (family Aleyrodidae). Bemisia tabaci transmits 111 of these species while Trialeurodes vaporariorum and T. abutilonia transmit three species each. B. tabaci and T. vaporariorum are present in the European–Mediterranean region, though the former is restricted in its distribution. Of the whitefly-transmitted virus species, 90% belong to the Begomovirus genus, 6% to the Crinivirus genus and the remaining 4% are in the Closterovirus, Ipomovirus or Carlavirus genera. Other named, whitefly-transmitted viruses that have not yet been ranked as species are also documented. The names, abbreviations and synonyms of the whitefly-transmitted viruses are presented in tabulated form together with details of their whitefly vectors, natural hosts and distribution. Entries are also annotated with references. Whitefly-transmitted viruses affecting plants in the European–Mediterranean region have been highlighted in the text.     

The competence of four thrips species to transmit and replicate four tospoviruses

The tospoviruses Tomato spotted wilt virus (TSWV), Tomato chlorotic spot virus (TCSV), Groundnut ringspot virus (GRSV) and Chrysanthemum stem necrosis virus (CSNV) are well-known pathogens on tomato in Brazil. The thrips species Frankliniella occidentalis , F. schultzei , Thrips tabaci and T. palmi were studied for their competence to transmit these tospoviruses. Frankliniella occidentalis transmitted all four tospoviruses with different efficiencies. Frankliniella schultzei transmitted TCSV, GRSV and CSNV. Although F. schultzei has been reported as a vector of TSWV, the F. schultzei population in the present study did not transmit the TSWV isolate used. A population of T. tabaci known to transmit Iris yellow spot virus (onion isolate) did not transmit any of the studied tospoviruses, and nor did T. palmi . Replication of these tospoviruses could be demonstrated by ELISA, not only in the thrips species that could transmit them, but also in those that could not. The results strongly suggest that competence to transmit is regulated not only by the initial amount of virus acquired and replication, but also by possible barriers to virus circulation inside the thrip's body.     

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.     

Replication of Tomato spotted wilt virus After Ingestion by Adult Thrips setosus is Restricted to Midgut Epithelial Cells

ABSTRACT If acquisition access feeding (AAF) is first given after adult eclosion, none of the nine thrips species able to serve as tospovirus vectors can become infective. The previous cellular investigations of this phenomenon, carried out only in Frankliniella occidentalis, suggested that infectivity was prevented because the type member of the tospoviruses, Tomato spotted wilt virus (TSWV), was unable to enter the midgut of adult thrips. The present study extends a cellular view of tospovirus-thrips interactions to a species other than the western flower thrips, F. occidentalis. Our findings show that TSWV enters and replicates within the midgut of adult Thrips setosus, but does not infect cells beyond the midgut epithelia. After AAF as adult, TSWV replicated in T. setosus midgut cells as indicated by significant increases in nucleocapsid (N) protein detected by double-antibody sandwich enzyme-linked immunosorbent assay, and the presence of inclusions containing the S RNA-encoded nonstructural and N proteins revealed by microscopic observations. Electron microscopic observations of adult insects showed that no infection occurred in cells beyond the midgut epithelia, and insects subsampled from the same cohorts could not transmit TSWV. In contrast, electron microscopy observations of larval T. setosus revealed that TSWV infected the midgut and muscle cells, and adult insects developing from these cohorts had infected salivary glands and were able to transmit TSWV. Mature virions were observed only in the salivary glands of adults developing from infected larvae. Our findings suggest that the barrier to infectivity in T. setosus adults differs from that shown for F. occidentalis adults.     

Increase of tospoviral diversity in Brazil with the identification of two new tospovirus species, one from chrysanthemum and one from zucchini

ABSTRACT During a survey conducted in several different regions of Brazil, two unique tospoviruses were isolated and characterized, one from chrysanthemum and the other from zucchini. The chrysanthemum virus displayed a broad host range, whereas the virus from zucchini was restricted mainly to the family Cucurbitaceae. Double-antibody sandwich-enzyme-linked immunosorbent assay and western immunoblot analyses demonstrated that both viruses were serologically distinct from all reported tospovirus species including the recently proposed peanut yellow spot virus and iris yellow spot virus (IYSV) species. The nucleotide sequences of the nucleocapsid (N) genes of both viruses contain 780 nucleotides encoding for deduced proteins of 260 amino acids. The N proteins of these two viruses displayed amino acid sequence similarities with the previously described tospovirus species ranging from 20 to 75%, but they were more closely related to each other (80%). Based on the biological and molecular features, these viruses are proposed as two new tospovirus species, designated chrysanthemum stem necrosis virus (CSNV) and zucchini lethal chlorosis virus (ZLCV). With the identification of CSNV and ZLCV, in addition to tomato spotted wilt virus, groundnut ring spot virus, tomato chlorotic spot virus, and IYSV, Brazil harbors the broadest spectrum of tospovirus species reported.     

Virus transmission by host-specific strains ofOlpidium bornovanus andOlpidium brassicae

Zoospores of 12 isolatesO. bornovanus from geographically diverse sites and representing the three host specific cucurbit strains were tested as vectors for seven viruses using watermelon bait plants and the in vitro acquisition method. All isolates of the cucumber, melon, and squash strains transmitted melon necrotic spot carmovirus (MNSV) and cucumber necrosis tombusvirus (CNV) but none transmitted petunia asteroid mosaic tombusvirus (PAMV) or tobacco necrosis necrovirus (TNV). The isolates varied as vectors of three other carmoviruses: cucumber leaf spot virus (CLSV); cucumber soil borne virus (CSBV); and squash necrosis virus (SqNV). All cucumber isolates transmitted CLSV and SqNV but not CSBV. Some of the melon isolates transmitted CLSV and SqNV but none transmitted CSBV. Two squash isolates transmitted CSBV and SqNV but not CLSV. Two isolates ofO. brassicae transmitted only TNV and a third did not transmit any of the viruses. The species of bait plant sometimes affected transmission. The most efficient vector strains ofO. bornovanus, as determined by reducing zoospores and virus in the inoculum, were the cucumber strain for CLSV; the cucumber strain for CNV if cucumber was the bait plant or melon strain if watermelon was the bait plant; and the squash strain for SqNV. The plurivorous strain ofO. brassicae was the most efficient vector of TNV.Olpidium bornovanus is the first vector reported for CSBV and is confirmed as a vector of SqNV. It is proposed that all carmoviruses may have fungal vectors.Ligniera sp. did not transmit any of the viruses in one attempt.Abbreviations CLSV cucumber leaf spot virus - CNV cucumber necrosis virus - CSBV cucumber soil borne virus - MNSV melon necrotic spot virus - PAMV petunia asteroid mosaic virus - SqNV squash necrosis virus - TNV tobacco necrosis virus - TBSV tomato bushy stunt virus     

昆虫与弹状病毒互作的研究进展

近年来宏基因组学研究的普及大大丰富了人们对RNA病毒多样性的认识,但对这些新发现病毒的生物学特性却所知甚少。本文围绕RNA病毒中一类重要的负单链RNA病毒--弹状病毒与其昆虫寄主互作的研究进行综述,总结已发现的弹状病毒及其昆虫寄主类型,共有20个属144种弹状病毒可以感染14个属的昆虫;根据已有的系统进化研究对弹状病毒的寄主起源进行推测;并以感染黑腹果蝇Drosophila melanogaster的sigma病毒(Drosophila melanogaster sigma virus,DMelSV)为主要对象,就弹状病毒引起的CO2麻痹致死症状以及昆虫寄主对其的免疫反应研究进行总结,而在对黑腹果蝇的研究中发现很多非经典免疫通路中的新抗病毒基因,暗示存在新的抗病毒免疫通路;通过飞虱、叶蝉与其传播的植物弹状病毒以及长须罗蛉Lutzomyia longipalpis与其传播的脊椎动物病毒的互作研究,发现Toll、IMD信号通路、细胞自噬及小RNA干扰(small interfering RNA,siRNA)通路等可能与昆虫对弹状病毒的免疫反应相关。昆虫是弹状病毒主要的寄主和媒介,也是病毒遗传多样性的储主,因此更好地研究和了解昆虫寄主与弹状病毒的相互关系,有助于病毒致病和传播机制以及昆虫抗病毒免疫机理的深入研究。     

Analysis of Iris yellow spot virus replication in vector and non‐vector thrips species

Iris yellow spot virus (IYSV, genus Tospovirus) is a viral disease of bulb and seed onion crops and is transmitted by Thrips tabaci. Foliage damage of up to 75% has been reported in Kenya and Uganda. In this study, the rate of IYSV replication in the larva, pupa and adult stages of T. tabaci and other non‐vector thrips species and colour forms such as Frankliniella occidentalis, F. schultzei (dark) and F. schultzei (pale) was evaluated by monitoring relative levels of nucleocapsid (N) and non‐structural (NSs) proteins using N‐ and NSs‐specific antibodies. The effect of IYSV replication on mortality of thrips was also determined. N protein levels increased in all three stages of IYSV‐fed T. tabaci, indicating replication of IYSV. In IYSV‐fed non‐vector thrips, the increase of N protein levels in the larval stage was lower than IYSV‐fed T. tabaci but higher than their healthy counterparts. The N protein levels did not increase at pupal and adult stages. NSs protein was not detected in first instar of either vector or non‐vector thrips species. After a 4 h post‐acquisition period, a significant increase in NSs proteins was only observed in IYSV‐fed T. tabaci, clearly differentiating vectors and non‐vectors of IYSV. IYSV replication did not influence the survival of the vector thrips species, T. tabaci populations or the non‐vector thrips species. This study indicates the effectiveness of monitoring non‐structural proteins such as NSs, compared to nucleocapsid proteins, for differentiating vectors and non‐vectors of IYSV.     

Diversity and detections of Phytophthora species from trade and non‐trade environments in Ireland

The genus Phytophthora is one of the genera of organisms that poses the most threat to plant health worldwide. Statutory monitoring for Phytophthora species focuses on the species regulated in the European Union and recommended for regulation by EPPO (Plant Health Directive 2000/29 EC and the EPPO A2 List). This research provides details of the Phytophthora species detected from trade and non‐trade environments in Ireland between 2013 and 2015. The results of statutory surveys for the regulated species Phytophthora ramorum, Phytophthora kernoviae and Phytophthora lateralis from 2003 to 2015 are also presented. Testing of more than 11 000 samples was carried out using morphological and/or DNA identification with specifically designed Phytophthora conserved primers. This led to the detection of 19 species and 3 informally designated taxa of Phytophthora, including 8 new records for Ireland. Eight species were found in both trade and non‐trade locations, and three informally designated taxa were also detected. Phytophthora ramorum was found on the most hosts (30 hosts), followed by Phytophthora syringae (6 hosts) and Phytophthora kernoviae (3 hosts). Rhododendron was the host on which Phytophthora species were most frequently detected (12 Phytophthora species). The role of the plant trade in spreading invasive Phytophthora species is discussed.     

Frankliniella occidentalis (Pergande) integrated pest management programs for fruiting vegetables in Florida

BACKGROUND: The spread of the western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), resulted in the worldwide destabilization of established integrated pest management programs for many crops. Efforts to control the pest and the thrips‐vectored tospoviruses with calendar applications of broad‐spectrum insecticides have been unsuccessful. The result has been a classic ‘3‐R’ situation: resistance to numerous insecticides; resurgence of the western flower thrips populations as a result of natural predators and native competitor thrips being eliminated; replacement by various other pests. This paper reports on integrated pest management programs for fruiting vegetables that are effective, economical, ecologically sound and sustainable. RESULTS: The components include the following: define pest status (economic thresholds); increase biotic resistance (natural enemies and competition); integrate preventive and therapeutic tactics (scouting, ultraviolet‐reflective technologies, biological control, compatible insecticides, companion plants and fertility); vertically integrate the programs with other pests; continually communicate latest science‐based management tactics with end‐users. CONCLUSION: These programs have been widely implemented in Florida and have significantly improved the management of western flower thrips and thrips‐transmitted viruses. Copyright © 2012 Society of Chemical Industry     

Biological and genetic diversity of plasmodiophorid-transmitted viruses and their vectors

About 20 species of viruses belonging to five genera, Benyvirus, Furovirus, Pecluvirus, Pomovirus and Bymovirus, are known to be transmitted by plasmodiophorids. These viruses have all positive-sense, single-stranded RNA genomes that consist of two to five RNA components. Three species of plasmodiophorids are recognized as vectors: Polymyxa graminis, P. betae, and Spongospora subterranea. The viruses can survive in soil within the long-lived resting spores of the vector. There are biological and genetic variations in both virus and vector species. Many of the viruses are causal agents of important diseases in major crops such as rice, wheat, barley, rye, sugar beet, potato, and groundnut. Control is dependent on the development of resistant cultivars. During the last half century, several virus diseases have rapidly spread worldwide. For six major virus diseases, we address their geographical distribution, diversity, and genetic resistance.     

Plant‐parasitic nematodes of potential phytosanitary importance,their main hosts and reported yield losses

The potential phytosanitary importance of all named plant‐parasitic nematode species was determined by evaluating available information on species characteristics, association with economically‐important crop hosts, and ability to act as vectors of viruses or form disease complexes with other pathogens. Most named species of plant‐parasitic nematodes (PPN) are poorly known, recorded from a single location only, not associated with economically‐important crops, and not known to be associated with other plant disease organisms. However, 250 species from 43 genera fulfilled one or more of the criteria to be considered to present a phytosanitary risk. The genera and number of species (in parentheses) considered as posing phytosanitary risk included: Achlysiella (1), Anguina (8), Aphasmatylenchus (1), Aphelenchoides (12), Aphelenchus (1), Belonolaimus (2), Bitylenchus (3), Bursaphelenchus (4), Cactodera (3), Ditylenchus (8), Dolichodorus (1), Globodera (3), Helicotylenchus (7), Hemicriconemoides (3), Hemicycliophora (3), Heterodera (25), Hirschmanniella (5), Hoplolaimus (5), Ibipora (3), Longidorus (10), Macroposthonia (2), Meloidogyne (38), Merlinius (3), Nacobbus (1), Neodolichodorus (2), Paralongidorus (2), Paratrichodorus (11), Paratylenchus (3), Pratylenchus (24), Punctodera (3), Quinisulcius (3), Radopholus (5), Rotylenchulus (3), Rotylenchus (1), Scutellonema (5), Sphaeronema (1), Subanguina (3), Trichodorus (5), Tylenchorhynchus (8), Tylenchulus (2), Vittatidera (1), Xiphinema (15) and Zygotylenchus (1). For each of the 250 species main hosts and yield loss estimates are provided with an extensive bibliography. Of the 250 species, only 126 species from 33 genera are currently listed as regulated pests in one or more countries worldwide. Almost all of these 250 species were also associated with economically important crops and some also acted as vectors for viruses.     

The transmission of geminiviruses by Bemisia tabaci

The cosmopolitan whitefly species, Bemisia tabaci (Gennadius) and Trialeurodes taporariorum (Westwood) have always been regarded as pests to a large range of worldwide crops. Both species are capable of transmitting plant viruses, with T. vaporariorum being the vector of only a few ‘clostero’-like viruses and B. tabaci the vector of viruses in several groups. The largest group of viruses transmitted by B. tabaci are the geminiviruses and B. tabaci is known to transmit around 60 members. Until recently, B. tabaci had been associated with only a limited range of host plants within any one region, although its total potential host range was large. Virus transmission was confined within the plant host range of each regional population of B. tabaci. The emergence of the polyphagous ‘B’ biotype of B. tabaci with its increased host range of more than 600 plant species, has resulted in geminiviruses infecting previously unaffected crops. As the ‘B’ biotype spreads further into Europe, European field and glasshouse crops have been shown to be susceptible to whitefly-transmitted viruses already endemic to other parts of the world. More than 20 colonies of B. tabaci, including both ‘B’ and non-‘B’ biotypes from disparate global locations have been compared for their ability to transmit more than 20 geminiviruses. All but two highly host-specific colonies were capable of transmitting most geminiviruses tested. However, some viruses were transmitted more efficiently than others. The virus coat protein or capsid is essential for vector recognition and transmission. By comparing transmissible viruses at the molecular level to viruses that are no longer whitefly-transmissible, the active epitope on the virus coat protein could be identified for designing future virus control strategies.     

Host plant influences life cycle,reproduction, feeding,and vector competence of Thrips palmi (Thysanoptera: Thripidae), a vector of tospoviruses

Phytoparasitica - Thrips palmi, an important pest of vegetables and ornamentals, transmits several tospoviruses. Effect of tospovirus on host plant and vector is well studied. Little is known about...