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ToolsDeng, Longji, Li, Yixuan, Tanizawa, Yasuhiro, Nakamura, Yasukazu, Kamikawa, Ryoma, Hopes, Amanda and Mock, Thomas (2025) A genetic transformation system for the heterotrophic diatom Nitzschia putrida (Bacillariophyceae). Journal of Phycology, 61 (5). pp. 1116-1123. ISSN 0022-3646
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Abstract
Diatoms are important primary producers in aquatic ecosystems. Most of them are photoautotrophs and have evolved to thrive under diverse environmental conditions from the poles to the tropics. However, some diatom species such as Nitzschia putrida have lost photosynthesis and have therefore become free-living secondary heterotrophs. Thus, these diatoms provide unique opportunities to study the evolutionary processes required to thrive without photosynthesis and independent of a resource-providing host. They may also provide a chassis for reverse engineering photosynthesis in eukaryotic organisms. Here, we have developed a genetic transformation system for N. putrida using a biolistic approach. By leveraging genome and transcriptome data, we identified the nicotinamide adenine dinucleotide hydride (NADH)-ubiquinone reductase complex 1 promoter as a robust candidate for driving transgene expression. Through Golden Gate Cloning, we engineered plasmids, including the selectable marker nourseothricin and the reporter eGFP. An evaluation of transformation efficiency confirmed the successful integration and expression of the transgenes. Fluorescence microscopy demonstrated the expression of eGFP in the transformed cell lines, which retained a growth phenotype similar to that of the wild type cells. Thus, our work in combination with the available genome and transcriptome of N. putrida enables reverse genetics with a free-living secondary heterotroph.
| Item Type: | Article |
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| Additional Information: | Funding information: LD, YL, AH, and TM acknowledge the School of Environmental Sciences for partial support. TM acknowledges funding from NERC, UK (NE/R000883/1). JSPS Grant-in-Aid for Challenging Research Exploratory (24K21929) awarded to RK; JSPS Grants-in-Aid for Scientific Research on Innovative Areas, Platform for Advanced Genome Science (16H06279) to R.K., Y.T., and Y.N. YL acknowledges the China Scholarship Council (Certification No. 202106330018) for funding through the International Cooperative Training Program. |
| Uncontrolled Keywords: | diatom,gene expression,molecular,aquatic science,plant science ,/dk/atira/pure/subjectarea/asjc/1100/1104 |
| Faculty \ School: | Faculty of Science > School of Environmental Sciences Faculty of Science > School of Biological Sciences University of East Anglia Research Groups/Centres > Theme - ClimateUEA |
| UEA Research Groups: | Faculty of Science > Research Groups > Environmental Biology Faculty of Science > Research Centres > Centre for Ecology, Evolution and Conservation Faculty of Science > Research Groups > Wolfson Centre for Advanced Environmental Microbiology Faculty of Science > Research Groups > Centre for Ocean and Atmospheric Sciences |
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| Depositing User: | LivePure Connector |
| Date Deposited: | 14 Nov 2025 16:30 |
| Last Modified: | 16 Nov 2025 07:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/100992 |
| DOI: | 10.1111/jpy.70070 |
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