DFRL, Mysore, Karnataka-570011, India.
J Sci Food Agric. 2011 Jul;91(9):1666-73. doi: 10.1002/jsfa.4365. Epub 2011 Mar 28.
The genus Fusarium comprises a diverse group of fungi including several species that produce mycotoxins in food commodities. In the present study, a multiplex PCR was standardised for the group-specific detection of fumonisin-producing and trichothecene-producing strains of Fusarium species. Primers for genus-level recognition of Fusarium spp. were designed from the internal transcribed spacer regions 1 and 2 of rDNA. Primers for group-specific detection were designed from the tri5 and tri6 genes involved in trichothecene biosynthesis and the fum1 and fum13 genes involved in fumonisin biosynthesis.
Among the various genera and their strains tested, all the 85 confirmed Fusarium strains were positive for rDNA gene and the rest stayed negative. From among the Fusarium strains, 15 had amplification for trichothecene- and 20 for fumonisin-encoding genes. All PCR positive trichothecene chemotypes of Fusarium species tested were positive for chemical analysis but in the case of fumonisins, of the 20 PCR positive cultures, only 13 showed positive for chemical analysis by HPTLC.
The assay described here provided a rapid and reliable detection of trichothecene- and fumonisin-producing Fusarium directly from natural food grains and the results were always comparable with a conventional HPTLC detection method. It can, therefore, be used by the food industry to monitor quality and safety.
镰刀菌属真菌种类繁多，包括一些能在食品中产生真菌毒素的物种。在本研究中，我们建立了一种多重 PCR 方法，用于特异性检测产伏马菌素和产单端孢霉烯族毒素的镰刀菌属真菌。用于镰刀菌属种水平鉴定的引物是根据 rDNA 的内部转录间隔区 1 和 2 设计的。用于特异性检测的引物是根据参与单端孢霉烯族生物合成的 tri5 和 tri6 基因以及参与伏马菌素生物合成的 fum1 和 fum13 基因设计的。
在所测试的各种属及其菌株中，所有 85 株确认为镰刀菌的菌株的 rDNA 基因均呈阳性，其余均为阴性。在镰刀菌菌株中，有 15 株扩增出了编码单端孢霉烯族毒素的基因，有 20 株扩增出了编码伏马菌素的基因。在所测试的镰刀菌属单端孢霉烯族毒素化学型中，所有 PCR 阳性菌株的化学分析结果均为阳性，但在伏马菌素方面，20 株 PCR 阳性培养物中，只有 13 株通过 HPTLC 化学分析呈阳性。
本研究中描述的方法可以快速可靠地从天然谷物中直接检测产伏马菌素和产单端孢霉烯族毒素的镰刀菌，其结果与常规 HPTLC 检测方法始终具有可比性。因此，它可以被食品行业用于监测质量和安全。
