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Three methods to inoculate Lycopersicon esculentum 'VF Pink' seedlings with tomato spotted wilt virus (TSWV) were compared. Treatments were 1) two inoculations by hand (rubbing leaves with a sterile cotton swab), 2) a single inoculation using a paint sprayer at 3.56 × 105 N· m-2, and 3) two spray inoculations. All three methods were effective (>95% infection) under moderate temperatures in the spring, but hand inoculation was not effective under hot conditions in the summer. In another experiment, spray inoculation was used to compare effects of light intensity and the leaf inoculated on susceptibility of L.. hirsutum PI 127826, L. pimpinellifoliom LA 1580 and `VF Pink' to TSWV isolate 85-9. All three genotypes were susceptible under full sun and 60% shade cloth in the greenhouse. Inoculation of youngest leaves produced the highest virus titer. Background optical density for noninoculated plants differed between lower and upper leaves in the ELISA assay.
Seedlings of eight accessions of L. hirsutum and susceptible L. esculentum `VF Pink' controls were spray inoculated twice in the greenhouse with tomato spotted wilt virus (TSWV) Arkansas 85-9. Plants lacking symptoms were reinoculated, then evaluated for TSWV by enzyme-linked immunosorbent assay (ELISA). Controls were consistently infected; sixty noninfected L. hirsutum were propagated by cuttings and inoculated with TSWV isolates T2 (lettuce), G-87 (gloxinia), 87-34 (tomato) and a mixture of the four isolates. All selections became infected in at least one test, but systemic infection was often delayed. Additional wild Lycopersicon species and numbers of accessions evaluated for resistance to TSWV include L. cheesmanii (9), L. chmielewskii (17), L. hirsutum (24), L. hirsutum f. glabratum (17), L. parviflorum (4) and L. pennellii (44). No new sources of strong resistance have been identified yet. Evaluation of additional species and accessions is continuing.
Latent infection of Blackberry yellow-vein associated virus (BYVaV) in `Chickasaw' blackberry has been reported. However, plants with characteristic leaf symptoms, such as vein yellowing, chlorotic mottling, and oak-leaf patterns, have tested positive for BYVaV using reverse-transcription polymerase chain reaction. Experiments were initiated to determine if the symptoms expressed in BYVaV infected `Chickasaw' were caused by mixed virus infections. BYVaV, a recently identified crinivirus, was evaluated for synergistic interactions with Tobacco ringspot virus (TRSV), Tomato ringspot virus (ToRSV), and Raspberry bushy dwarf virus (RBDV). `Chickasaw' blackberry plants infected with BYVaV (single infection) were used as receptor plants to establish mixed virus infections with TRSV and ToRSV transmitted by nematodes and RBDV transmitted by bottle grafts. Characteristic symptoms of multi-virus infection will be presented and discussed.