Authors
A. M.
Abouzid
,
J.
Freitas-Astua
, and
D. E.
Purcifull
,
Department of Plant Pathology, University of Florida, Gainesville 32611-0680
;
J. E.
Polston
,
Gulf Coast Research and Education Center, Bradenton, FL 34203
;
K. A.
Beckham
,
W. E.
Crawford
,
M. A.
Petersen
, and
B.
Peyser
,
Department of Plant Pathology, University of Florida
;
C.
Patte
,
Gulf Coast Research and Education Center
; and
E.
Hiebert
,
Department of Plant Pathology, University of Florida
ABSTRACT
Polyclonal rabbit antisera were produced to the coat protein of Bean golden mosaic virus Brazil isolate (BGMV), Cabbage leaf curl virus (CabLCV), Tomato yellow leaf curl virus (TYLCV), and Tomato mottle virus (ToMoV), all expressed in Escherichia coli by the pETh expression vector. The expressed coat protein of each virus was purified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for use as an immunogen. The antisera to BGMV, CabLCV, TYLCV, and ToMoV reacted in indirect (plate-trapping) enzyme-linked immunosorbent assay (ELISA) with extracts from begomovirus-infected tissue. The antisera to BGMV, CabLCV, TYLCV, and ToMoV also reacted specifically with the test begomovirus antigens in leaf imprint blots and Western blots. The CabLCV and TYLCV antisera were used to detect Bean golden yellow mosaic virus antigens by immunogold labeling of thin sections of infected bean tissues. In tissue blot immunoassays, the TYLCV antiserum reacted well with TYLCV antigens but not with ToMoV antigens, while CabLCV antiserum reacted well with ToMoV antigens and weakly with TYLCV antigens. The results indicate that polyclonal antisera prepared to expressed begomovirus coat proteins were useful for the detection of begomoviruses in an array of assays.
