Detection of Lettuce Infectious Yellow Virus (LIYV) in Greenhouse and Field Inoculated Plots Using an Indirect Enzyme-linked Immunosorbent Assay (Indirect ELISA)

Abstract

Lettuce infectious yellows virus (LIYV), a recently recognized plant virus, causes dramatic yellowing symptoms and severe diseases in a wide range of vegetable crops in Arizona, adjacent southwestern states and Mexico. Until now, the only available diagnostic method was a time-consuming bioassay that used the insect vector to transmit the virus, with subsequent manipulation of indicator plants. A rapid, sensitive diagnostic technique (termed an indirect enzyme-linked immunoassay, called indirect ELISA) system was developed to detect lettuce infectious yellows virus (LIYV) in infected plant material. A virus specific antibody was made to viral capsid protein which was purified by polyacrylamide gel electrophoresis. The indirect ELISA system was optimized and used to detect viral antigen in greenhouse-inoculated melons. The system was subsequently adapted to detect LIYV in symptomatic and asymptomatic weed and cultivated plant species collected from infected fields near Yuma and in central Arizona. The indirect ELISA system described here allows for the detection of approximately 100 ng of virus per well. The LIYV was detectable in symptomatic (but not in asymptomatic) leaves of melon plants infected with the virus. In contrast, the virus could be detected in both symptomatic and symptomless cheeseweed plants collected in the field. The optical density readings for infected weed species were generally lower than those for cultivated species, such as melons, lettuce, and spinach, suggesting that there is less virus in the weed hosts tested than in infected, cultivated hosts.