Scott Adkins

Tomato spotted wilt virus—positive steps towards negative success

Abstract Taxonomy: Tomato spotted wilt virus (TSWV) is the type member of the plant-infecting Tospovirus genus in the family Bunyaviridae, a large group of predominantly vertebrate- and insect-infecting RNA viruses. Physical properties: Virions are 80-120-nm pleomorphic particles with surface projections composed of two viral glycoproteins, G1 and G2 (Fig. 1). Virion composition is 5% nucleic acid, 70% protein, 5% carbohydrate and 20% lipid. The genome consists of three negative or ambisense ssRNAs designated S (2.9 kb), M (4.8 kb) and L (8.9 kb), with partially complementary terminal sequences that allow the RNA to adopt a pseudocircular or panhandle conformation. Each genomic RNA is encapsidated by multiple copies of the viral nucleocapsid (N) protein to form ribonucleoprotein structures also known as nucleocapsids. The nucleocapsids are enclosed in a host-derived membrane bilayer along with an estimated 10-20 copies of the L protein, the putative RNA-dependent RNA polymerase. Hosts: Over 800 plant species, both dicots and monocots, in more than 80 plant families are susceptible to TSWV (Goldbach and Peters, 1994). The Solanaceae and Compositae families contain the largest numbers of susceptible plant species (Prins and Kormelink, 1998). TSWV also replicates in its insect vector, thrips (Thysanoptera: Thripidae) (Ullman et al., 1993; Wijkamp et al., 1993). Useful web site: http://www4.ncbi.nlm.nih.gov/ICTVdb/ICTVdB/11050003.htm.

Role of the Insect Supervectors Bemisia tabaci and Frankliniella occidentalis in the Emergence and Global Spread of Plant Viruses

Emergence of insect-transmitted plant viruses over the past 10-20 years has been disproportionately driven by two so-called supervectors: the whitefly, Bemisia tabaci, and the Western flower thrips, Frankliniella occidentalis. High rates of reproduction and dispersal, extreme polyphagy, and development of insecticide resistance, together with human activities, have made these insects global pests. These supervectors transmit a diversity of plant viruses by different mechanisms and mediate virus emergence through local evolution, host shifts, mixed infections, and global spread. Associated virus evolution involves reassortment, recombination, and component capture. Emergence of B. tabaci-transmitted geminiviruses (begomoviruses), ipomoviruses, and torradoviruses has led to global disease outbreaks as well as multiple paradigm shifts. Similarly, F. occidentalis has mediated tospovirus host shifts and global dissemination and the emergence of pollen-transmitted ilarviruses. The plant virus-supervector interaction offers exciting opportunities for basic research and global implementation of generalized disease management strategies to reduce economic and environmental impacts.

A natural M RNA reassortant arising from two species of plant- and insect-infecting bunyaviruses and comparison of its sequence and biological properties to parental species

Reassortment allows multicomponent viruses to exchange genome segments, a process well-documented in the vertebrate- and arthropod-infecting members of the family Bunyaviridae but not between distinct species of the plant- and insect-infecting members of the genus Tospovirus. Genome sequence comparisons of a virus causing severe tospovirus-like symptoms in Florida tomato with Groundnut ringspot virus (GRSV) and Tomato chlorotic spot virus (TCSV) demonstrated that reassortment has occurred, with the large (L) and small (S) RNAs coming from GRSV and the medium (M) RNA coming from TCSV (i.e. L(G)M(T)S(G)). Neither parental genotype is known to occur in the U.S. suggesting that L(G)M(T)S(G) was introduced as a reassortant. L(G)M(T)S(G) was transmitted by western flower thrips (Frankliniella occidentalis [Pergande]), and was not able to overcome the Sw5 resistance gene of tomato. Our demonstration of reassortment between GRSV and TCSV suggests caution in defining species within the family Bunyaviridae based on their ability to reassort.

Identification and Characterization of a Novel Whitefly-Transmitted Member of the Family Potyviridae Isolated from Cucurbits in Florida

ABSTRACT A novel whitefly-transmitted member of the family Potyviridae was isolated from a squash plant (Cucurbita pepo) with vein yellowing symptoms in Florida. The virus, for which the name Squash vein yellowing virus (SqVYV) is proposed, has flexuous rod-shaped particles of approximately 840 nm in length. The experimental host range was limited to species in the family Cucurbitaceae, with the most dramatic symptoms observed in squash and watermelon, but excluded all tested species in the families Amaranthaceae, Apocynaceae, Asteraceae, Chenopodiaceae, Fabaceae, Malvaceae, and Solanaceae. The virus was transmitted by whiteflies (Bemisia tabaci) but was not transmitted by aphids (Myzus persicae). Infection by SqVYV induced inclusion bodies visible by electron and light microscopy that were characteristic of members of the family Potyviridae. Comparison of the SqVYV coat protein gene and protein sequences with those of recognized members of the family Potyviridae indicate that it is a novel member of the genus Ipomovirus. A limited survey revealed that SqVYV also was present in watermelon plants suffering from a vine decline and fruit rot recently observed in Florida and was sufficient to induce these symptoms in greenhouse-grown watermelon, suggesting that SqVYV is the likely cause of this disease.

Biological and molecular characterization of a novel tobamovirus with a unique host range

Tobamoviruses are among the best characterized and most studied plant viruses. Three subgroups of tobamoviruses correspond to viral genome sequence and host range to include those viruses infecting (i) solanaceous plants, (ii) brassicas, or (iii) cucurbits or legumes. We isolated a virus from Florida landscape plantings of the malvaceous plant hibiscus (Hibiscus rosasinensis) that appears to be a tobamovirus based upon its virion morphology, genome organization, and coat protein sequence. The experimental host range of this virus included five malvaceous species but excluded all tested brassica, cucurbit, and legume species and 12 of the 19 solanaceous species tested. The unique host range and comparison of coat protein gene and protein sequences with those of recognized tobamoviruses indicate that this is a novel to-bamovirus. A limited survey revealed that this virus is widespread in hibiscus and related species in the Florida landscape.

Squash vein yellowing virus Detection Using Nested Polymerase Chain Reaction Demonstrates that the Cucurbit Weed Momordica charantia Is a Reservoir Host

Squash vein yellowing virus (SqVYV) is a recently described ipomovirus from cucurbits in Florida that induces the relatively unusual symptoms in watermelon of plant death and fruit rind necrosis and discoloration, commonly known in Florida as watermelon vine decline. In this report, SqVYV infection of Momordica charantia (Balsam-apple), a common cucurbit weed, collected in 2005 and 2007 from within or adjacent to fields of declining watermelon, is demonstrated through the use of nested polymerase chain reaction (PCR). M. charantia plants located in or around fallow watermelon fields between spring and fall 2007 watermelon crops were also infected with SqVYV, indicating that this weed can serve as an oversummering host for this virus. Furthermore, whiteflies were able to acquire SqVYV from infected M. charantia and transmit it to squash and watermelon. Nested PCR was 10 to 1,000 times more sensitive than non-nested PCR for SqVYV detection in several cucurbit hosts, including M. charantia and watermelon. Melothria pendula (creeping cucumber), another common cucurbit weed, was experimentally infected with SqVYV. These results suggest that improved management of M. charantia and other cucurbit weeds needs to be incorporated into watermelon vine decline management plans to reduce sources of SqVYV and other cucurbit viruses.

Emergence of Groundnut ringspot virus and Tomato chlorotic spot virus in Vegetables in Florida and the Southeastern United States

Groundnut ringspot virus (GRSV) and Tomato chlorotic spot virus (TCSV) are two emerging tospoviruses in Florida. In a survey of the southeastern United States, GRSV and TCSV were frequently detected in solanaceous crops and weeds with tospovirus-like symptoms in south Florida, and occurred sympatrically with Tomato spotted wilt virus (TSWV) in tomato and pepper in south Florida. TSWV was the only tospovirus detected in other survey locations, with the exceptions of GRSV from tomato (Solanum lycopersicum) in South Carolina and New York, both of which are first reports. Impatiens (Impatiens walleriana) and lettuce (Lactuca sativa) were the only non-solanaceous GRSV and/or TCSV hosts identified in experimental host range studies. Little genetic diversity was observed in GRSV and TCSV sequences, likely due to the recent introductions of both viruses. All GRSV isolates characterized were reassortants with the TCSV M RNA. In laboratory transmission studies, Frankliniella schultzei was a more efficient vector of GRSV than F. occidentalis. TCSV was acquired more efficiently than GRSV by F. occidentalis but upon acquisition, transmission frequencies were similar. Further spread of GRSV and TCSV in the United States is possible and detection of mixed infections highlights the opportunity for additional reassortment of tospovirus genomic RNAs.

Presence of P1b and absence of HC-Pro in Squash vein yellowing virus suggests a general feature of the genus Ipomovirus in the family Potyviridae

The genus Ipomovirus is one of six currently recognized genera in the family Potyviridae. The complete nucleotide sequence of Squash vein yellowing virus (SqVYV), a putative ipomovirus recently described in Florida, has been determined. The 9836 nt SqVYV genomic RNA [excluding the poly(A) tail] has one large open reading frame encoding a single polyprotein of 3172 amino acids, typical of the genome organization for most members in the family Potyviridae. The 10 mature proteins predicted to be derived from the SqVYV polyprotein include P1a and P1b but no HC-Pro, similar to Cucumber vein yellowing virus (CVYV) but different from Sweet potato mild mottle virus (SPMMV), both recognized members of the genus Ipomovirus. Phylogenetic analysis of these proteins supports classification of SqVYV as a novel species within the genus Ipomovirus. However, the similar genome organization strategy of SqVYV and CVYV, which differs from that of SPMMV, indicates that the taxonomy of the genus Ipomovirus needs to be re-examined and a new genus created within the family Potyviridae to accommodate the observed discrepancies in ipomovirus genome organization.

Determination of complete nucleotide sequence of Hibiscus latent Singapore virus: Evidence for the presence of an internal poly(A) tract

Summary.We have sequenced the complete genome of a hibiscus-infecting tobamovirus, Hibiscus latent Singapore virus (HLSV). The experimental host range of HLSV is similar to that of another distinct species of hibiscus infecting tobamovirus, Hibiscus latent Fort Pierce virus (HLFPV). The genomic structure of HLSV is similar to other tobamoviruses in general. It consists of a 5′ untranslated region (UTR), followed by ORFs encoding for a 128 kDa protein and a 186 kDa readthrough protein, a 30 kDa movement protein (MP), 18 kDa coat protein (CP) and a 3′ UTR. The unique feature of HLSV is the presence of a poly(A) tract within its 3′ UTR. In our previous work, we have reported MP and CP sequences of HLSV and its phylogenetic analysis. Here we report the complete nucleotide sequence of HLSV, phylogenetic analysis of the nucleotide and amino acid sequences of 128/186 kDa ORFs and the presence of a uniquely located poly(A) tract within the 3′ UTR.

Key West Nightshade, a New Experimental Host for Plant Viruses

Key West nightshade (Solanum bahamense) is a perennial solanaceous weed found in the extreme southern portion of Florida. It can be propagated by seed and cuttings and is absent from the noxious weed lists of all U.S. states. Its susceptibility to five viruses common to Florida was evaluated by mechanical inoculation of leaves with Tomato spotted wilt virus (TSWV), Tobacco mosaic virus (TMV), Pepper mild mottle virus (PMMoV), Cucumber mosaic virus (CMV), and a putative tobamovirus recently isolated from hibiscus in Florida (HV). TSWV induced chlorotic rings on inoculated leaves and mosaic and malformation of uninoculated leaves. CMV induced necrotic local lesions on inoculated leaves. No symptoms were observed following inoculation with TMV, PMMoV, or HV. TSWV, TMV, and PMMoV systemically infected S. bahamense as determined by the use of enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, viral-associated double-stranded RNA analysis, and/or indicator hosts. Active growth of infected plants continued for 7 months following inoculation, making S. bahamense suitable for long-term maintenance of viruses in planta. We suggest that S. bahamense may be a useful host for virus culture collections and for studies involving large numbers of virus isolates where fresh, infected tissue is continuously required.