The Plant Pathology Journal 2005;21(4):301-309.
Published online December 31, 2005.
Partial Purification of Lectin from Mycoparasitic Species of Trichoderma
Tanuja Singh, Ratul Saikia, Dilip K. Arora
Trichoderma species/isolates exhibited varied degree of agglutination on sclerotial (Sc) and hyphal (Hy) surface of Macrophomina phaseolina. The agglutination efficiencies on Sc and Hy ranged from 11 to 57%. Isolates of T. harzianum (Th) and T. viride (Tv) showed greater agglutination on Sc (23-57%) and Hy (16-47%). Different enzymes (trypsin, pepsin, proteinase k, α-chymotrypsin, lyticase and glucosidase) and inhibitors (tunicamycin,cycloheximide, brefeldin A, sodium azide,dithiothreitol and SDS) reduced the agglutination potential of conidia of Th-23/98 and Tv-25/98; however, the extent of response varied greatly in different treatments.Different fractions of Th-23/98 and Tv-25/98 exhibited haemagglutinating reaction with human blood group A, B, AB and O. Haemagglutinating activity was inhibited by different sugars and glycoproteins tested. Crude haemagglutinating protein from outer cell wall protein fraction of Th-23/98 and Tv-25/98 were eluted on Sephadex G-100 column. Initially Th-23/98 and Tv- 25/98 exhibited two peaks showing no agglutination activity; however, lectin activity was detected in the third peak. Similar to crude lectin, the purified lectin also exhibited haemagglutinating activity with different erythrocyte source. SDS-PAGE analysis of partially purified lectin revealed single band with an estimated molecular mass of 55 and 52 kDa in Th-23/98 and Tv- 25/98, respectively. Trypsin, chymotrypsin and b-1,3- glucanase totally inhibited lectin activity. Similarly,various pH also affected the haemagglutinating activity of Th-23/98 and Tv-25/98. From the present observations, it can be concluded that the recognition/attachment of mycoparasite (T. harzianum and T. viride) to the host surface (M. phaseolina) may be most likely due to lectincarbohydrate interaction.
Key Words: agglutination, Lectin, Macrophomina phaseolina, recognition, Trichoderma
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