Abstract
Various modifications of the tissue-blot immunoassay (TBIA) for the detection of barley yellow dwarf virus (BYDV, luteovirus) were compared. Similar results were obtained by using three different labelled molecules; goat anti-rabbit antibodies conjugated to alkaline phosphatase, protein A conjugated with alkaline phosphatase and goat anti-rabbit antibodies conjugated with colloidal gold. Blocking the nitrocellulose membrane with polyvinyl alcohol for 1 min was effective and allowed the procedure to be shortened by one hour. TBIA was sensitive enough to detect BYDV in old dry tissue wich had been soaked in water for 1 h.
BYDV was monitored by TBIA in wheat, oat and barley after inoculation at heading, flowering and grain filling growth stages. The later the inoculation date, the greater the chance of detecting the virus in stem bases rather than in the upper part of the stem. The later the inoculation the less virus moved, from the inoculated tiller to other tillers of the same plant.
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References
Clark MF and Adams AN (1977) Characteristics of the microplate method enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology 34: 475–483
Comeau A (1987) Effects of BYDV inoculation at various dates for oats, barley, wheat, rye and triticale. Phytoprotection 68: 97–109
Hsu YH (1984) Immunogold for detection of antigen on nitrocellulose paper. Analytical Biochemistry 142: 221–225
Hsu HT and Lawson RH (1991) Direct tissue blotting for detection of tomato spotted wilt virus in Impatiens. Plant Disease 75: 292–295
Lin NS, Hsu YH and Hsu HT (1990) Immunological detection of plant viruses and a mycoplasmalike organism by direct tissue blotting on nitrocellulose membranes. Phytopathology 80: 824–828
Makkouk KM, Comeau A and St-Pierre CA (1994) Screening for barley yellow dwarf luteovirus resistance in barley on the basis of virus movement. Journal of Phytopathology. In press
Miranda PV, Brandelli A and Tezon JG (1993) Instantaneous blocking for immunoblots. Analytical Biochemistry 209: 376–377
Shine JM and Comstock JC (1993) Digital image analysis system for determining tissue-blot immunoassay results for ratoon stunting disease of sugarcane. Plant Disease 77: 511–513
Van Regenmortel MHV (1982) Serology and Immunochemistry of Plant Viruses. Academic Press Inc., New York, 302 pp
Zagula KR, Barbara DJ, Fulbright DW and Lister RM (1990) Evaluation of three ELISA methods as alternatives to ISEM for detection of the wheat spindle streak mosaic strain of wheat yellow mosaic virus. Plant Disease 74: 974–978
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Makkouk, K.M., Comeau, A. Evaluation of various methods for the detection of barley yellow dwarf virus by the tissue-blot immunoassay and its use for virus detection in cereals inoculated at different growth stages. Eur J Plant Pathol 100, 71–80 (1994). https://doi.org/10.1007/BF01871967
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DOI: https://doi.org/10.1007/BF01871967