Identification and Functional Analysis of Two Mitoferrins, CsMIT1 and CsMIT2, Participating in Iron Homeostasis in Cucumber
Abstract
:1. Introduction
2. Results
2.1. Identification and Expression Analysis of CsMIT1 and CsMIT2
2.2. Mitochondrial Localization of CsMIT1 and CsMIT2 in Arabidopsis thaliana Protoplasts
2.3. Effect of CsMIT1 and CsMIT2 Expression on the Metal-Sensitive Yeast Mutants
2.4. Effect of CsMIT1 and CsMIT2 Expression on the Cytosolic and Mitochondrial Fe Content in Δmrs3Δmrs4 Cells
3. Discussion
4. Materials and Methods
4.1. Plant Material and Growth Conditions
4.2. RNA Extraction and Real-Time PCR
4.3. Cloning of CsMIT1 and CsMIT2
4.4. Yeast Strains, Media and Fluorescence Imaging
4.5. Metal Sensitivity Spot Assays
4.6. Cytosolic Gentisate 1,2-Dioxygenase Assays
4.7. Preparation of Mitochondrial Fraction from Yeast Cells
4.8. Determination of Mitochondrial Heavy Metal Content
4.9. Protein Determination
4.10. Transformation of Arabidopsis thaliana Protoplasts and Confocal Imaging
4.11. Bioinformatics and Accession Numbers
4.12. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Małas, K.; Kabała, K. Identification and Functional Analysis of Two Mitoferrins, CsMIT1 and CsMIT2, Participating in Iron Homeostasis in Cucumber. Int. J. Mol. Sci. 2023, 24, 5050. https://doi.org/10.3390/ijms24055050
Małas K, Kabała K. Identification and Functional Analysis of Two Mitoferrins, CsMIT1 and CsMIT2, Participating in Iron Homeostasis in Cucumber. International Journal of Molecular Sciences. 2023; 24(5):5050. https://doi.org/10.3390/ijms24055050
Chicago/Turabian StyleMałas, Karolina, and Katarzyna Kabała. 2023. "Identification and Functional Analysis of Two Mitoferrins, CsMIT1 and CsMIT2, Participating in Iron Homeostasis in Cucumber" International Journal of Molecular Sciences 24, no. 5: 5050. https://doi.org/10.3390/ijms24055050