The ZRT, IRT-like protein (ZIP) family exists in many species and plays an important role in many biological processes, but little is known about ZIP genes in Aspergillus oryzae. Here, 10 ZIP genes in A. oryzae were identified and these were classified into four groups based on phylogenetic analysis. The structures of these AoZip genes were determined, which indicated a great divergence of AoZip members from different groups. Synteny analysis revealed that AoZip7, AoZip8, and AoZip10 are conserved among Aspergillus species. We also found that the promoter regions of AoZip2, AoZip7, AoZip8, and AoZip10 contain multiple conserved response elements. Expression analysis revealed that AoZips exhibited different expression patterns in response to different metal treatments. Moreover, overexpression and RNA-interference (RNAi) of AoZip2 led to a decrease in mycelium growth diameter and inhibited conidia formation. AoZip2 overexpression and RNAi strains showed distinct sensitivity to severely Zn/Mn-depleted stress. In addition, kojic acid production was markedly lower in AoZip2 overexpression and RNAi strains than in the control strains, and the expression of kojA, kojR, and kojT was down-regulated in AoZip2 overexpression and RNAi strains. This study provides new insights into our understanding of ZIP genes and lays a foundation for further investigation of their roles in Aspergillus oryzae.

中文翻译：

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ZRT，IRT样蛋白（ZIP）家族基因的鉴定和表征表明，它们参与了米曲霉的生长和曲酸的生产。

ZRT，IRT样蛋白（ZIP）家族存在于许多物种中，并在许多生物学过程中起着重要作用，但是对于米曲霉中的ZIP基因知之甚少。在这里，鉴定了米曲霉中的10个ZIP基因，并且根据系统发育分析将它们分为四类。确定了这些AoZip基因的结构，这表明来自不同组的AoZip成员差异很大。同源性分析显示，AoZip7，AoZip8和AoZip10在曲霉菌种中是保守的。我们还发现，AoZip2，AoZip7，AoZip8和AoZip10的启动子区域包含多个保守的响应元件。表达分析表明，AoZips对不同的金属处理表现出不同的表达模式。此外，AoZip2的过表达和RNA干扰（RNAi）导致菌丝体生长直径的减少和分生孢子的形成。AoZip2过表达和RNAi菌株对严重缺锌/缺锰的胁迫表现出明显的敏感性。另外，在AoZip2过表达和RNAi菌株中曲酸的产量显着低于对照菌株，并且在AoZip2过表达和RNAi菌株中kojA，kojR和kojT的表达下调。这项研究为我们对ZIP基因的理解提供了新的见解，并为进一步研究其在米曲霉中的作用奠定了基础。在AoZip2过表达和RNAi菌株中曲酸的产量显着低于对照菌株，并且在AoZip2过表达和RNAi菌株中kojA，kojR和kojT的表达下调。这项研究为我们对ZIP基因的理解提供了新的见解，并为进一步研究其在米曲霉中的作用奠定了基础。在AoZip2过表达和RNAi菌株中曲酸的产量显着低于对照菌株，并且在AoZip2过表达和RNAi菌株中kojA，kojR和kojT的表达下调。这项研究为我们对ZIP基因的理解提供了新的见解，并为进一步研究其在米曲霉中的作用奠定了基础。