Defence Food Research Laboratory, Microbiology Devision, Sidartha Nagar, Mysore, Karnataka, India.
Braz J Microbiol. 2013 Oct 30;44(2):401-6. doi: 10.1590/S1517-83822013000200009. eCollection 2013.
Hundred Fusarium culmorum strains, isolated from freshly harvested maize grain samples from Southern parts of India, were incubated in czapek-dox medium and analyzed for trichothecene (DON/NIV) production. The mPCR assay was standardized targeting trichothecene metabolic pathway genes viz., Tri6, Tri7, Tri13 for detection of trichothecene (DON/NIV) chemotypes and rDNA gene for specific detection of F. culmorum species. Primers for targeted genes were designed and used to predict whether these isolates could produce deoxynivalenol/nivalenol, 94 isolates were able to produce DON/NIV by mPCR assay. Chemical analysis of DON/NIV was carried out for mPCR positive isolates by high performance-thin layer chromatography (HPTLC). To check the practical usefulness of developed mPCR assay, 150 field samples of maize were evaluated and results were compared with conventional HPTLC method. Out of 150 samples, 34% samples stayed as a positive for NIV contamination whereas 44% were found to have deoxynivalenol contamination. Moreover, mPCR results are equivocally matched with the HPTLC chemical analysis for field samples. Chemotyping of F. culmorum isolates were reported for the first time from India, and highlights the important potential of F. culmorum to contaminate maize with DON/NIV.
从印度南部新收获的玉米籽粒样本中分离出的 100 株镰刀菌,在察氏培养基中培养,并分析其是否产生三萜烯(DON/NIV)。通过 mPCR 检测方法,对三萜烯代谢途径基因(Tri6、Tri7、Tri13)进行了标准化,以检测三萜烯(DON/NIV)化学型,并用 rDNA 基因特异性检测镰刀菌属物种。设计了针对目标基因的引物,用于预测这些分离株是否能够产生脱氧雪腐镰刀菌烯醇/雪腐镰刀菌烯醇,通过 mPCR 检测,94 株分离株能够产生 DON/NIV。对 mPCR 阳性分离株进行了 DON/NIV 的化学分析,采用高效薄层色谱法(HPTLC)。为了检验所开发的 mPCR 检测方法的实际应用价值,对 150 份玉米田间样本进行了评估,并将结果与传统的 HPTLC 方法进行了比较。在 150 个样本中,有 34%的样本被检测出 NIV 污染呈阳性,而有 44%的样本被检测出脱氧雪腐镰刀菌烯醇污染。此外,mPCR 结果与田间样本的 HPTLC 化学分析结果一致。这是首次从印度报道镰刀菌属菌株的化学型,突出了镰刀菌属菌株对玉米 DON/NIV 污染的重要潜在风险。