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ARS Home » Southeast Area » Fort Pierce, Florida » U.S. Horticultural Research Laboratory » Subtropical Plant Pathology Research » Research » Publications at this Location » Publication #347454
Research Project: Mitigating High Consequence Domestic, Exotic, and Emerging Diseases of Fruits, Vegetables, and Ornamentals

Location: Subtropical Plant Pathology Research

Title: Integrating local lesion assays with conventional RT-PCR for detection of interspecies tospovirus reassortants and mixed tospovirus infections

Author
item TANTIWANICH, Y - Department Of Agriculture - Thailand
item CHIEMSOMBAT, P - Kasetsart University
item NAIDU, R.A. - Washington State University
item Adkins, Scott

Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/14/2017
Publication Date: 4/1/2018
Citation: Tantiwanich, Y., Chiemsombat, P., Naidu, R., Adkins, S.T. 2018. Integrating local lesion assays with conventional RT-PCR for detection of interspecies tospovirus reassortants and mixed tospovirus infections. Plant Disease. 102:715-719.

Interpretive Summary: Field symptoms of the three tospoviruses Tomato spotted wilt virus (TSWV), Groundnut ringspot virus (GRSV) and Tomato chlorotic spot virus (TCSV) are indistinguishable in most host plants. Commercially available TSWV immunoassay reagents cross react with GRSV and TCSV complicating reliable identification of these three species. All currently known isolates of GRSV in the U.S. are reassortants containing one genomic RNA segment derived from TCSV further confounding identification. To address these challenges, we developed and validated specific primers for conventional RT-PCR detection of TSWV, GRSV and TCSV.

Technical Abstract: Tomato spotted wilt virus (TSWV) has historically been the major tospovirus species present in North America. Recent emergence of Groundnut ringspot virus (GRSV) and Tomato chlorotic spot virus (TCSV) in Florida and the Caribbean has complicated reliable identification of tospoviruses in this region. Field symptoms of these three tospoviruses are indistinguishable in most host plants and commercially available TSWV lateral flow immunoassay reagents cross react with GRSV and TCSV leading to incorrect diagnoses of GRSV or TCSV as TSWV. Reliable diagnosis of TSWV, GRSV and TCSV is further confounded by the fact that all currently known isolates of GRSV in the U.S. are reassortants containing one genomic RNA segment derived from TCSV. To address these practical challenges, we developed and validated genome segment-specific primers for conventional RT-PCR detection of the L, M and S RNA segments of TSWV, GRSV and TCSV. When used in conjunction with local lesion-passaged virus isolates, the genome segment-specific RT-PCR assays developed in this study will facilitate high-throughput screening of plant or thrips samples for interspecies reassortants in epidemiological studies and reliable identification of these three tospovirus species in mixed infections commonly observed in the field.