Richardson K E, Hagler W M, Hamilton P B
Appl Environ Microbiol. 1984 Apr;47(4):643-6. doi: 10.1128/aem.47.4.643-646.1984.
Three methods for detecting toxigenic fusaria in culture were compared by using known producers of zearalenone, zearalenol, T-2 toxin, and deoxynivalenol. Moist, autoclaved rice cultures of known toxigenic isolates grown in 20-ml tubes yielded oily extracts containing compounds which interfered with qualitative and quantitative analysis for the mycotoxins. Vermiculite moistened with nutrient broth in 20-ml tubes yielded a much cleaner extract. Growing the fungi on a liquid medium required a shorter incubation period, but yields of T-2 toxin and deoxynivalenol were low and variable, and the method required greater space in the incubator. Screening of the extracts by thin-layer chromatography with colorimetric spray reagents to detect the presence of these toxins permitted reduction in the number of extracts quantified by the more lengthy gas-liquid chromatographic method. Culturing in nutrient broth on vermiculite in tubes coupled to a qualitative screen before quantitation proved to be a convenient, inexpensive, and relatively rapid method that enabled reliable screening of a large number of Fusarium isolates for toxin production as compared with prior methods.
通过使用已知的玉米赤霉烯酮、玉米赤霉醇、T-2毒素和脱氧雪腐镰刀菌烯醇产生菌,比较了三种在培养物中检测产毒镰刀菌的方法。在20毫升试管中培养的已知产毒分离株的潮湿、经高压灭菌的水稻培养物产生了油性提取物,其中含有干扰霉菌毒素定性和定量分析的化合物。在20毫升试管中用营养肉汤湿润的蛭石产生了更纯净的提取物。在液体培养基上培养真菌所需的培养时间较短,但T-2毒素和脱氧雪腐镰刀菌烯醇的产量较低且不稳定,并且该方法需要在培养箱中占用更大空间。用比色喷雾试剂通过薄层色谱法对提取物进行筛选以检测这些毒素的存在,从而减少了通过更冗长的气液色谱法进行定量的提取物数量。与先前的方法相比,在试管中的蛭石上用营养肉汤培养并在定量前进行定性筛选被证明是一种方便、廉价且相对快速的方法,能够可靠地筛选大量镰刀菌分离株的毒素产生情况。