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• W935051539 abstract "The toxigenicity of isolates of Fusarium for chickpea, Cicer arietinum, the third mostimportant legume crop in the world was studied. Fungi were grown in liquid culture and theculture filtrates assayed on cells isolated from leaflets of the plant. One isolate, designatedFOC 5, produced cultures that were predominately red (70-80% of the cultures). When theculture filtrates of all isolates over time were assayed, the red cultures of FOC 5 were muchmore toxic than those of the other isolates and were also about 10 times more toxic than thecolourless cultures of FOC 5. Toxic titres of the red FOC 5 cultures peaked at 12 days whengrown at 20°C. The toxin from these red cultures were purified by solvent partitioning, solidphase extraction (SPE), thin layer chromatography (TLC) and high performance liquidchromatography (HPLC) using the assay to monitor the stages in purification.Shaking of culture filtrates of FOC 5 with ethyl acetate resulted in about half the toxicactivity (50-55%) partitioning into the organic phase and 25-30% remaining in the aqueousphase. The activity of the aqueous phase was lost on freeze-drying suggesting a volatilecompound. When the ethyl acetate phase was dissolved in aqueous acetonitrile and applied toC18 SPE cartridges, about 9% was not adsorbed and 35% could be eluted with methanol.Greater affinity was shown for cyano SPE cartridges with 6% not adsorbed and 45 %recoverable by elution in acetonitrile. Attempts at purification of the toxin(s) of adsorbed andnon-adsorbed fractions from these reversed phase cartridges by HPLC did not yield pureproducts.Recovery of activity of the ethyl acetate phase from flash chromatography on silicagel was 61-110%. However, HPLC demonstrated that several compounds were present in theactive fractions.Separation of components of the ethyl acetate phase or the fraction adsorbed by cyanocartridges of culture filtrates by TLC on silica gel rather than using SPE, flash or reversedphase HPLC was more successful. Red bands corresponding to the active compound werescraped from TLC plates and eluted in acetonitrile. HPLC of the eluents on a cyano columnwith 10% acetonitrile as the mobile phase demonstrated a single homogeneous peak withabsorption maxima of 224 and 281 nm. The purified fraction is, at the time of writing, beingstudied by Professor Mike Beale at Rothamsted Research using nuclear magnetic resonancetechniques in order to determine its structure.Four other isolates, identified by the International Crops Research Institute for theSemi-Arid Tropics as F. oxysporum f. sp. ciceri did not produce the red, toxic compound,throwing doubt on the correct identification of the isolates. When the sequences of ribosomalDNA of all five isolates were determined, the isolate that produced the red toxic compoundmost closely matched Fusarium acutatum (99%), in a BLAST search and this accorded withits morphology. A BLAST search showed that three of the other isolates matched thesequence of cotton pathogen, F. oxysporum f. sp. vasinfectum (100%, 100% and 97%) andone closely matched F. oxysporum f. sp. vanillae (99%) These results suggest that a reevaluationof the taxonomy of Fusarium species causing wilt of chickpea is required." @default.
• W935051539 created "2016-06-24" @default.
• W935051539 creator A5081801696 @default.
• W935051539 date "2004-01-01" @default.
• W935051539 modified "2023-09-28" @default.
• W935051539 title "Toxigenicity of Fusarium species causing wilt of chickpea" @default.
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