Schmidt-Heydt Markus, Geisen Rolf
Federal Research Centre for Nutrition and Food, Location Karlsruhe, Haid-und-Neu-Str. 9 76131 Karlsruhe, Germany.
Int J Food Microbiol. 2007 Jun 30;117(2):131-40. doi: 10.1016/j.ijfoodmicro.2007.01.014. Epub 2007 Mar 30.
A microarray, which covers most of the known relevant mycotoxin biosynthesis genes, has been developed. The microarray carries oligonucleotides of the fumonisin, the aflatoxin, the ochratoxin, the trichothecene (type A and B) and the patulin biosynthesis pathways. For trichothecene producing Fusaria the biosynthesis cluster of trichothecene producing Fusarium sporotrichioides (type A) and of Gibberrella zeae (type B, teleomorph of F. graminearum) have been spotted. The aflatoxin cluster carries oligonucleotides specific for Aspergillus flavus. The ochratoxin pattern is specific for ochratoxin A producing Penicillia, the fumonisin cluster is specific for G. moniliformis (teleomorph of F. verticillioides) and the patulin genes have been obtained from Penicillium expansum. The microarray is designed in a way that newly identified pathway genes can be added easily at any time. The microarray was used to detect the activation of all gene clusters under conditions conducive for mycotoxin biosynthesis. According to the results the obtained signals were specific under the hybridization conditions used and only insignificant cross-hybridizations occurred. The microarray was used to demonstrate differences in mycotoxin pathway gene expressions after growth on various media for trichothecene and ochratoxin A biosynthesis. It was used further to study and compare the expression kinetics of the trichothecene biosynthesis genes of Fusarium on different trichothecene supporting media. An expression pattern indicative for trichothecene biosynthesis could be identified.
一种覆盖了大多数已知相关霉菌毒素生物合成基因的微阵列已经研制出来。该微阵列带有伏马菌素、黄曲霉毒素、赭曲霉毒素、单端孢霉烯族毒素(A 型和 B 型)以及展青霉素生物合成途径的寡核苷酸。对于产生单端孢霉烯族毒素的镰刀菌,已点样了产单端孢霉烯族毒素的拟分枝孢镰刀菌(A 型)和禾谷镰刀菌(B 型,F. graminearum 的有性型)的生物合成簇。黄曲霉毒素簇带有对黄曲霉特异的寡核苷酸。赭曲霉毒素模式对产赭曲霉毒素 A 的青霉特异,伏马菌素簇对串珠镰刀菌(F. verticillioides 的有性型)特异,展青霉素基因取自扩展青霉。该微阵列的设计方式使得新鉴定出的途径基因可在任何时候轻松添加。该微阵列用于检测在有利于霉菌毒素生物合成的条件下所有基因簇的激活情况。根据结果,在所使用的杂交条件下获得的信号具有特异性,仅发生了不显著的交叉杂交。该微阵列用于证明在用于单端孢霉烯族毒素和赭曲霉毒素 A 生物合成的各种培养基上生长后霉菌毒素途径基因表达的差异。它还被进一步用于研究和比较镰刀菌在不同支持单端孢霉烯族毒素合成的培养基上单端孢霉烯族毒素生物合成基因的表达动力学。可以鉴定出一种指示单端孢霉烯族毒素生物合成的表达模式。