An obviously unhealthy plant identified as Tragopogon mirus Ownbey (remarkable goatsbeard) was sent for diagnosis to the Division of Plant Industry (DPI), Gainesville, FL in May of 2008. T. mirus is a recently formed allotetraploid that has T. dubius Scop. and T. porrifolius L. (goatsbeard or salsify) as parents. The parents (family Asteraceae) are diploid and originate from Eurasia. They were introduced to the northwest United States in the early 1900s. The allotetraploid T. mirus, which does not occur in Eurasia, was discovered in 1949 and named in 1950. It has been found in the northwest states of Washington and Idaho. It has also been found in Arizona (4). The plant sent to the DPI was grown in a greenhouse for research purposes at the Botany Department of the University of Florida (Alachua County). Symptoms exhibited on the leaves included mottling, chlorotic and necrotic spots, and mild distortion. Epidermal leaf strips from a mottled leaf were stained with the Orange-Green protein stain and Azure A nucleic acid stain (1). With a light microscope, granular inclusions typical for Tomato spotted wilt virus (TSWV) (1) were seen in leaf strips from both stains. The remainder of the leaf was ground in buffer and tested serologically for TSWV by TSWV-specific ImmunoStrips (Agdia, Elkhart, IN). The ImmunoStrip was positive for the presence of TSWV. This test was confirmed by double-antibody sandwich-ELISA using antiserum and conjugate for TSWV (Agdia). Further serological testing of other Tragopogon species with similar symptoms growing in the same greenhouse revealed that T. miscellus (another recently formed allotetraploid found in the northwestern United States; parents T. dubius and T. pratensis), T. dubius, T. porrifolius, and T. pratensis were also infected with TSWV. Total RNA was extracted from symptomatic leaves of T. mirus, T. dubius, T. porrifolius, and T. miscellus. Reverse transcription-PCR was performed with universal tospovirus primers BR60 and BR65 that amplify part of the nucleocapsid protein gene (2). Target amplicons of 454 bp were produced for all four samples. The PCR product from T. porrifolius was cloned and sequenced. The resulting sequence (GenBank Accession No. FJ655913) shows high homology, 98%, to several isolates of the Tomato spotted wilt virus deposited in the GenBank (Accession Nos. AY870391, AY744477, and AF020659). T. porrifolius has been reported to be naturally infected with TSWV in Italy (3); however, to our knowledge, this is the first report of this virus in the allotetraploids T. mirus and T. miscellus and in the diploids T. dubius and T. pratensis. This report adds five new Asteraceae weeds to the list of possible reservoirs of TSWV in the United States. References: (1) J. R. Edwardson and R. G. Christie. Univ. Fla. Inst. Food Agric. Sci. Bull. 894. 1996. (2) M. Eiras et al. Fitopatol. Bras. 26:170, 2001. (3) G. Parrella et al. J. Plant Pathol. 85:227. 2003. (4) D. E. Soltis et al. Biol. J. Linn. Soc. 82:2004.
Characterization and Establishment of a Recombinase Polymerase Amplification Assay for Rapid Detection of Tomato Spotted Wilt Virus in Pepper
