Microbiology and Kluyver Centre for Genomics of Industrial Fermentations, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Environ Microbiol. 2011 Jan;13(1):216-225. doi: 10.1111/j.1462-2920.2010.02322.x. Epub 2010 Aug 16.
Colonization of a substrate by fungi starts with the invasion of exploring hyphae. These hyphae secrete enzymes that degrade the organic material into small molecules that can be taken up by the fungus to serve as nutrients. We previously showed that only part of the exploring hyphae of Aspergillus niger highly express the glucoamylase gene glaA. This was an unexpected finding since all exploring hyphae are exposed to the same environmental conditions. Using GFP as a reporter, we here demonstrate that the acid amylase gene aamA, the α-glucuronidase gene aguA, and the feruloyl esterase gene faeA of A. niger are also subject to heterogenic expression within the exploring mycelium. Coexpression studies using GFP and dTomato as reporters showed that hyphae that highly express one of these genes also highly express the other genes encoding secreted proteins. Moreover, these hyphae also highly express the amylolytic regulatory gene amyR, and the glyceraldehyde-3-phosphate dehydrogenase gene gpdA. In situ hybridization demonstrated that the high expressers are characterized by a high 18S rRNA content. Taken together, it is concluded that two subpopulations of hyphae can be distinguished within the exploring mycelium of A. niger. The experimental data indicate that these subpopulations differ in their transcriptional and translational activity.
真菌对基质的定殖始于探索性菌丝的入侵。这些菌丝会分泌酶,将有机物质降解成小分子,真菌可以将这些小分子吸收作为营养物质。我们之前曾表明,黑曲霉的只有部分探索性菌丝高度表达葡糖淀粉酶基因 glaA。这是一个出乎意料的发现,因为所有探索性菌丝都暴露在相同的环境条件下。在这里,我们使用 GFP 作为报告基因,证明黑曲霉的酸性淀粉酶基因 aamA、α-葡萄糖醛酸酶基因 aguA 和阿魏酸酯酶基因 faeA 也在探索性菌丝内表现出异质表达。使用 GFP 和 dTomato 作为报告基因的共表达研究表明,高度表达这些基因之一的菌丝也高度表达其他编码分泌蛋白的基因。此外,这些菌丝还高度表达淀粉酶调控基因 amyR 和甘油醛-3-磷酸脱氢酶基因 gpdA。原位杂交表明,高表达者的 18S rRNA 含量较高。综上所述,可以得出结论,在黑曲霉的探索性菌丝内可以区分出两种菌丝亚群。实验数据表明,这些亚群在转录和翻译活性上存在差异。