C6-like锌指蛋白介导氨基酸代谢在嗜盐曲霉Aspergillus montevidensis耐盐中的分子机制

嗜盐真菌独特的耐盐特性使其在盐碱环境修复中具有重要应用价值。目前真菌积累耐渗透压物质的耐盐机制研究主要集中在碳素代谢方面。我们发现嗜盐曲霉(Aspergillus montevidensis ZYD4)特异积累鸟氨酸、丙氨酸及天冬氨酸等代谢物同菌株耐盐密切相关，且介导氨基酸代谢的C6-like锌指蛋白编码基因(Amf1)显著上调表达，但其相关调控机制不明。本项目以嗜盐曲霉(ZYD4)为供试菌株，拟开展以下研究：构建Amf1敲除突变株(Δamf1)及过表达菌株；研究Amf1的高盐诱导表达特征；分析不同菌株形态及耐盐性的变化，认识C6-like锌指蛋白在嗜盐曲霉耐盐中的细胞功能；阐明C6-like锌指蛋白调控“精氨酸和脯氨酸代谢”、“丙氨酸/天冬氨酸和谷氨酸代谢”基因的转录机制及含氮代谢物的积累规律。研究成果对阐明嗜盐真菌耐盐新机制有重要意义，也为真核生物耐盐调控及盐碱环境修复提供新思路。

The unique halotolerant characteristics of halophilic fungi make them important for the remediation of saline-alkali environments. At present, studies on compatible solutes for enhancing the salt-tolerant properties of fungi are mainly focused on carbon metabolism. We found that the accumulation of specific metabolites in Aspergillus montevidensis ZYD4, such as ornithine, alanine, and aspartic acid was closely correlated with the increased salt tolerance of the fungus, and that the Amf1 gene encoding the C6-like zinc finger transcription factor, a key regulatory protein in mediating amino acid metabolism, was significantly up-regulated. The associated regulatory mechanism remains unknown. This project intends to carry out the following research using the fungus ZYD4 as a test strain: construct the Amf1 gene knock-out mutant (Δamf1) and its overexpression strain; study expression patterns of the Amf1 gene under high-salt conditions; analyze the changes in morphology and salt-tolerant properties of different strains to understand the cellular functions of C6-like zinc finger protein during the halophilic fungus ZYD4's adaptation to saline environment; elucidate the transcription mechanism of genes and the accumulation characteristics of nitrogen-containing metabolites in the pathways of "arginine and proline metabolism" and "alanine/aspartic acid and glutamic acid metabolism", which are putatively regulated by the C6-like zinc finger regulatory protein. The research results are of great significance because they reveal a new mechanism underlying the salt adaptation process of halophilic fungi, and provide new ideas for further research on salt-tolerant regulation in eukaryotes and for the remediation of saline-alkali environments.