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In order to understand the molecular mechanisms underlying transmission of Spiroplasma citri by the leafhopper Circulifer haematoceps, we screened leafhopper proteins as putative S. citri-binding molecules using a spiroplasma overlay assay of protein blots (Far-western assay). Insect proteins were separated by one- or two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis, blotted, and probed with S. citri proteins. In this in vitro assay, we found that spiroplasma proteins exhibited affinity for seven leafhopper proteins. The interactions between S. citri proteins and insect proteins with molecular masses of 50 and 60 kDa were found to be sugar sensitive. These insect proteins were identified as high mannose N-glycoproteins, which support an interaction of glycoprotein-lectin type with S. citri proteins. Lectin detection in S. citri has revealed only one protein of 24 kDa. Using a leafhopper protein …