Stable nuclear transformation of Gonium pectorale
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BACKGROUND: Green algae of the family Volvocaceae are a model lineage for studying the molecular evolution of multicellularity and cellular differentiation. The volvocine alga Gonium is intermediate in organizational complexity between its unicellular relative, Chlamydomonas, and its multicellular relatives with differentiated cell types, such as Volvox. Gonium pectorale consists of approximately 16 biflagellate cells arranged in a flat plate. The detailed molecular analysis of any species necessitates its accessibility to genetic manipulation, but, in volvocine algae, transformation procedures have so far only been established for Chlamydomonas reinhardtii and Volvox carteri. RESULTS: Stable nuclear transformation of G. pectorale was achieved using a heterologous dominant antibiotic resistance gene, the aminoglycoside 3'-phosphotransferase VIII gene (aphVIII) of Streptomyces rimosus, as a selectable marker. Heterologous 3'- and 5'-untranslated flanking sequences, including promoters, were from Chlamydomonas reinhardtii or from Volvox carteri. After particle gun bombardment of wild type Gonium cells with plasmid-coated gold particles, transformants were recovered. The transformants were able to grow in the presence of the antibiotic paromomycin and produced a detectable level of the AphVIII protein. The plasmids integrated into the genome, and stable integration was verified after propagation for over 1400 colony generations. Co-transformants were recovered with a frequency of approximately 30-50% when cells were co-bombarded with aphVIII-based selectable marker plasmids along with unselectable plasmids containing heterologous genes. The transcription of the co-transformed, unselectable genes was confirmed. After heterologous expression of the luciferase gene from the marine copepod Gaussia princeps, which was previously engineered to match the codon usage in C. reinhardtii, Gonium transformants show luciferase activity through light emission in bioluminescence assays. CONCLUSION: Flanking sequences that include promoters from C. reinhardtii and from V. carteri work in G. pectorale and allow the functional expression of heterologous genes, such as the selectable marker gene aphVIII of S. rimosus or the co-transformed, codon-optimized G. princeps luciferase gene, which turned out to be a suitable reporter gene in Gonium. The availability of a method for transformation of Gonium makes genetic engineering of this species possible and allows for detailed studies in molecular evolution using the unicellular Chlamydomonas, the 16-celled Gonium, and the multicellular Volvox.
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Nuclear transformation has been achieved in a number of other green algae species, such as members of the Chlorella and volvox genuses.
“while nuclear transformation has been achieved in a number of other green algae species, such as members of the Chlorella and volvox genuses [99,100].”
From Biofuels from algae: Challenges and potential (2010) · cites this paper as [99]
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Bhat, R. A., Singh, D. V., Tonelli, F. M. P., & Hakeem, K. R. (2022) · Springer eBooks · doi:10.1007/978-3-030-94074-4_1
Yamashita, S. (2023) · PLANT MORPHOLOGY · doi:10.5685/plmorphol.35.69
- Authors
- Lerche, K., & Hallmann, A.
- Affiliations
- Bielefeld University (DE)
- Corresponding author
- Hallmann, Armin
- ORCID
- Hallmann, Armin
- Venue
- BMC Biotechnology
- Published
- 2009
- Volume
- 9
- Issue
- 1
- Pages
- 64
- ISSN
- 1472-6750 (ISSN-L)
- PMC
- PMC2720962
- OpenAlex
- W2025282208
- Cited by
- 58
- References
- 48
- Citations by year
- 2025: 1 · 2024: 2 · 2023: 3 · 2022: 5 · 2021: 3 · 2020: 3 · 2019: 3 · 2018: 2 · 2017: 3 · 2016: 6 · 2015: 7 · 2014: 1 · 2013: 3 · 2012: 7
- FWCI
- 2.15
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- BMC Biotechnology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- PUB – Publications at Bielefeld University (Bielefeld University) (submittedVersion, other-oa)
- Publikationen an der Universität Bielefeld (Universität Bielefeld) (submittedVersion, other-oa)
- Funders
- University of Maryland, Baltimore County
- Universität Regensburg
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protist diversity and phylogeny
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- Gonium pectorale, volvocine algae, Volvocaceae, Streptomyces rimosus, co-transformation, heterologous expression, Gaussia princeps luciferase, codon optimization
- MeSH terms
- Chlorophyta, Base Sequence, Genetic Engineering, Molecular Sequence Data, Phylogeny, Promoter Regions, Genetic, Streptomyces, Transformation, Genetic, Genes, Reporter, Kanamycin Kinase, 5' Flanking Region, 3' Flanking Region, DNA, Algal
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