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Identification and characterization of Nramp transporter AoNramp1 in Aspergillus oryzae
Junxia Fan1, Huanxin Zhang2, Yuzhen Li1
1Jiangxi Key Laboratory of Bioprocess Engineering and Co-Innovation Center for In-Vitro Diagnostic Reagents and Devices of Jiangxi Province, College of Life Sciences, Jiangxi Science and Technology Normal University, Nanchang, 330013 China.
Abstract:
The Nramp (natural resistance-associated macrophage protein) family of genes has been identified and characterized widely in many species. However, the Nramp genes and their characterizations have not been reported for Aspergillus oryzae. Here, only one Nramp gene AoNramp1 in A. oryzae genome was identified. Phylogenetic analysis revealed that AoNramp1 is not clustered with Nramps from yeast genus. Expression analysis showed that the transcript level of AoNramp1 was strongly induced under both Zn/Mn-replete and -deplete conditions. The GUS-staining assay indicated that the expression of AoNramp1 was strongly induced by Zn/Mn. Moreover, the AoNramp1 deletion and overexpression strains were constructed by the CRISPR/Cas9 system and A. oryzae amyB promoter, respectively. Phenotypic analysis showed that overexpression and deletion of AoNramp1 caused growth defects under Zn/Mn-deplete and -replete conditions, including mycelium growth and conidia formation. Together, these findings provide valuable information for further study on the biological roles of AoNramp1 in A. oryzae.
Supplementary Information:
The online version contains supplementary material available at 10.1007/s13205-021-02998-z.
Insights
Researchers identified and characterized the natural resistance-associated macrophage protein 1 (AoNramp1) gene in Aspergillus oryzae. AoNramp1 plays a crucial role in metal ion homeostasis, impacting fungal growth and development under varying nutrient conditions.
Area of Science:
- Microbiology
- Molecular Biology
- Fungal Genetics
Background:
- The Nramp gene family is vital for metal ion transport across species.
- Characterization of Nramp genes in Aspergillus oryzae was previously lacking.
- This study focuses on the identification and function of AoNramp1 in A. oryzae.
Purpose of the Study:
- To identify and characterize the Nramp gene in Aspergillus oryzae.
- To investigate the role of AoNramp1 in metal ion (Zn/Mn) homeostasis.
- To analyze the impact of AoNramp1 expression on fungal growth and development.
Main Methods:
- Bioinformatic analysis for Nramp gene identification in A. oryzae.
- Phylogenetic analysis of the identified AoNramp1 gene.
- Gene expression analysis under varying Zn/Mn conditions using quantitative PCR and GUS staining.
- CRISPR/Cas9 system for gene deletion and amyB promoter for gene overexpression.
- Phenotypic analysis of deletion and overexpression strains.
Main Results:
- A single Nramp gene, AoNramp1, was identified in the A. oryzae genome.
- AoNramp1 expression is significantly induced by both zinc/manganese-replete and -deplete conditions.
- Overexpression and deletion of AoNramp1 led to growth defects under different metal ion concentrations.
- AoNramp1 is involved in regulating mycelium growth and conidia formation.
Conclusions:
- AoNramp1 is a critical metal ion transporter in Aspergillus oryzae.
- The gene plays a significant role in fungal adaptation to metal availability.
- Findings provide a foundation for understanding AoNramp1's biological functions in A. oryzae.
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