Tissue Blot Immunoassay for Detection of Tomato spotted wilt virus in Ranunculus asiaticus and Other Ornamentals

A tissue blot immunoassay (TBIA) was developed to detect Tomato spotted wilt virus (TSWV) in Ranunculus asiaticus tubers and other ornamentals. TBIA was comparable to double-antibody sandwich enzyme-linked immunosorbent assay for accuracy and reliability. A nondestructive sampling method was used with R. asiaticus tubers to determine: (i) the relationship between tuber infection and size; (ii) the distribution of TSWV in tubers; and (iii) the relationship between tuber infection and tuber germination. Small tubers had a higher percentage (44%) of infection than large tubers (19%). When destructive sampling was acceptable, the central stem tissue was the most reliable to test. TSWV infection was associated with a significant reduction of tuber germination. Among the tubers that tested positive for TSWV infection, 48% of those that germinated produced foliage in which TSWV was detected. The remaining 52% of the infected tubers planted that germinated developed into plants that were asymptomatic and in which TSWV was not detectable after germination. Only 4% of tubers that tested negative produced infected plants after germination. Our results indicate that TBIA can be used in TSWV management programs to identify infected plants and to index tuber crops.