Figure 2
Analysis of acid-hydrolyzed extracts from tomato leaves. (A) TLC of acid-hydrolyzed materials (aglycones, FIII) extracted from tomato leaves harvested 0, 48 and 96 hours after challenge with B. cinerea. Lanes: C, control plant; B, BTP1-treated plants. Samples (35 ÎĽl corresponding to 35 mg leaf fresh weight) were applied and the plate (silica gel) was developed with a mixture of dichloromethane: hexane: methanol (6:4:1, v/v/v). A conidial suspension of Cladosporium cucumerinum was then sprayed on the dried plate and fungitoxic zones (white spots) were revealed after incubation for 48 h. (B) HPLC profiles obtained for analyses a sample of 15 ÎĽl of fungitoxic zone extract scrapped from TLC (corresponding to lane B 48 hours after infection) was injected on a C-18 reverse-phase column and material was eluted at various flow rates with a gradient of acetonitrile from 5% to 95%. Chromatograms represent max plots calculated from the maximum absorbance between 200 nm and 500 nm of eluting material.