Ilgen Peter, Hadeler Birgit, Maier Frank J, Schäfer Wilhelm
Biocenter Klein-Flottbek, Molecular Phytopathology and Genetics, University of Hamburg, Ohnhorststrasse 18, 22609 Hamburg, Germany.
Mol Plant Microbe Interact. 2009 Aug;22(8):899-908. doi: 10.1094/MPMI-22-8-0899.
The fungal pathogen Fusarium graminearum is the most common agent of Fusarium head blight (FHB) in small grain cereals and cob rot of maize. The threat posed by this fungus is due to a decrease in yield and, additionally, mycotoxin contamination of the harvested cereals. Among the mycotoxins, trichothecenes influence virulence of F. graminearum in a highly complex manner that is strongly host- as well as chemotype-specific. The factors inducing mycotoxin production during plant infection are still unknown. To evaluate the induction of the trichothecene pathway, the green fluorescence protein (GFP) gene was fused to the promoter of the TRI5 gene coding for the trichodiene synthase and integrated into the genome by homologous integration. The resulting mutant contains a fully functional TRI5 gene ensuring virulence on wheat and exhibits GFP driven by the endogenous TRI5 promoter. We are now able to monitor the induction of trichothecenes under real-time conditions. To localize the fungus in the plant tissue, the dsRed gene was integrated under constitutive control of the glycerol-3-phosphate dehydrogenase (gpdA) promoter. We are now able to show that, first, induction of GFP as well as trichothecene production in the reporter strain reflects TRI5 induction and trichothecene production in the wild type; second, expression of TRI5 is inducible during growth in culture; and, third, trichothecene production is not uniformly induced during the onset of infection but is tissue specific during fungal infection of wheat.
真菌病原体禾谷镰刀菌是引起小粒谷物赤霉病(FHB)和玉米穗腐病的最常见病原体。这种真菌造成的威胁在于产量下降,此外,收获的谷物还会受到霉菌毒素污染。在霉菌毒素中,单端孢霉烯族毒素以高度复杂的方式影响禾谷镰刀菌的毒力,这种方式具有很强的宿主特异性和化学型特异性。植物感染期间诱导霉菌毒素产生的因素仍然未知。为了评估单端孢霉烯族毒素途径的诱导情况,将绿色荧光蛋白(GFP)基因与编码单端孢霉烯合酶的TRI5基因的启动子融合,并通过同源整合将其整合到基因组中。所得突变体含有一个功能完全正常的TRI5基因,确保对小麦具有毒力,并表现出由内源性TRI5启动子驱动的GFP。我们现在能够在实时条件下监测单端孢霉烯族毒素的诱导情况。为了在植物组织中定位真菌,将dsRed基因整合到甘油-3-磷酸脱氢酶(gpdA)启动子的组成型控制之下。我们现在能够证明,首先,报告菌株中GFP的诱导以及单端孢霉烯族毒素的产生反映了野生型中TRI5的诱导和单端孢霉烯族毒素的产生;其次,TRI5的表达在培养生长过程中是可诱导的;第三,在感染开始时,单端孢霉烯族毒素的产生并非均匀诱导,而是在小麦真菌感染期间具有组织特异性。