Trapping of Tomato yellow leaf curl virus (TYLCV) and other plant viruses with a GroEL homologue from the whitefly Bemisia tabaci.
To avoid destruction in the haemolymph of their vector, many plant circulative viruses interact with GroEL homologues produced by insect endosymbiotic bacteria. We have exploited this phenomenon to devise tools allowing trapping of plant viruses by either GroEL purified from the whitefly Bemisia tabaci or by whitefly GroEL over-expressed in E. coli. PCR tubes or 96-well plates coated with a GroEL preparation were incubated with cleared sap of virus infected plant leaves or insect vectors. GroEL-bound viruses were then identified by PCR or RT-PCR using virus-specific primers or by ELISA with virus specific antibodies. In this way Tomato yellow leaf curl virus (TYLCV) - a whitefly-transmitted geminivirus - was detected in plant sap, in extracts of leaf squashes and in homogenates of individual viruliferous whiteflies. Anti-GroEL antibody prevented TYLCV binding to GroEL. GroEL-bound virus was also detected by ELISA. GroEL was much more potent in binding TYLCV than commercial anti-TYLCV antibodies. In addition to several other geminiviruses, these procedures allowed detecting a variety of RNA viruses such as Cucumber mosaic virus (CMV), Prune dwarf virus (PDV) and Tomato spotted wilt (TSWV), but not Potato virus X and Potato virus Y (PVX and PVY), Grapevine leafroll-associated viruses (GLRV) and Tobacco mosaic virus (TMV). Predictions pertaining to viruses that do, or do not bind to GroEL, and applications in plant virus diagnosis, are presented.
The Minerva Otto Warburg Center for Agricultural Biotechnology, Faculty of Agriculture, The Hebrew University of Jerusalem, Rehovot, Israel.
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F
Akad
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Dotan
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Czosnek
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D018834:Chaperonin 60 / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D017791:Geminiviridae / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006430:Hemiptera / Q000737:chemistry / Q000235:genetics*; D007303:Insect Vectors / Q000235:genetics / Q000821:virology*; D010936:Plant Extracts / Q000032:analysis; D018515:Plant Leaves / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D012328:RNA Viruses / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000378:metabolism
D000914:Antibodies, Viral; D018834:Chaperonin 60; D017931:DNA Primers; D010936:Plant Extracts; D011994:Recombinant Proteins