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The effectiveness of aqueous extracts of various medicinal plants in detoxification of aflatoxin B1 (AFB1) was tested in vitro by thin-layer chromatography and enzyme-linked immunosorbent assay (ELISA). Among the different plant extracts, the leaf extract of Vasaka (Adhatoda vasica Nees) showed the maximum degradation of AFB1 (≥ 98%) after incubation for 24h at 37 °C. The aflatoxin detoxifying activity of the A. vasica leaf extract was significantly reduced by heating to 100 °C for 10 min or autoclaving at 121 °C for 20 min. Dialysis had no effect on aflatoxin detoxifying ability of A. vasica extract and the dialyzed extract showed similar level of detoxification of AFB1 as that of the untreated extract. A time course study of aflatoxin detoxification by A. vasica extract showed that 69% of the toxin was degraded within 6h and ≥ 95% degradation was observed after 24h of incubation. Detoxification of AFB1 by A. vasica extract was further confirmed by liquid chromatography-mass spectrometry (LC-MS) analysis. Phytochemical analysis revealed the presence of alkaloids in methanolic extract of A. vasica leaves. A partially purified alkaloid from A. vasica leaves by preparative TLC exhibited strong AFB1 detoxification activity.

The effect of soil application of biocontrol agents (Pseudomonas fluorescens, Trichoderma viride and T. harzianum) in combination with chitin on induction of phenolics and defense enzymes in coconut roots infected with Ganoderma lucidum, the causal agent of Ganoderma disease, was investigated. Soil application of these biocontrol formulations in combination with chitin induced a significant increase in the activities of peroxidase (PO), polyphenol oxidase (PPO), phenylalanine ammonia-lyase (PAL), chitinase and beta-1,3-glucanase in the G. lucidum infected palms. Activities of both PAL and PO reached maximum levels within 3 d while the activity of PPO reached the maximum level 6 d after application of a mixture of P. fluorescens, T. viride and chitin. Isozyme analysis revealed that unique PO3 and PPO2 isozymes were induced in coconut palms treated with P. fluorescens + T. viride + chitin. Accumulation of phenolics was recorded 3 d after treatment and reached maximum levels 9 d after treatment application. Activity of chitinase was significantly increased from the third day after treatment imposition and continued to increase up to 9 to 12 d in all treatments. Chitinase isozyme analysis revealed that a unique Chit3 isoform was induced in coconut roots treated with P. fluorescens + T. viride + chitin. The beta-1,3-glucanase activity was maximum 9 d after treatment application. The mechanisms by which P. fluorescens + T. viride + chitin reduced the incidence of Ganoderma disease in coconut may be related to its ability to induce defense mechanisms in coconut palms.

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