Genetic transformation methods for diatom Nitzschia captiva: New tools to better understand dinotom endosymbiosis

Genetic transformation methods for diatom Nitzschia captiva: New tools to better understand dinotom endosymbiosis

Endosymbiosis is a widespread and ecologically significant phenomenon in the marine environment. How these endosymbiotic partners evolve into an organism with a new organelle is still mostly unknown and requires investigation into modern symbioses. Dinotoms, dinoflagellates with evolutionarily inter...

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Bibliographic Details
Published in:Algal research (Amsterdam) Vol. 72; p. 103136
Main Authors: Sprecher, Brittany N., Buck, Jochen M., Ropella, L. Loraine, Ramsperger, Annette, Kroth, Peter G., Yamada, Norico
Format: Journal Article
Language:English
Published: Elsevier B.V 01-05-2023
Subjects:
Cryopreservation
Diatoms
Dinoflagellates
Endosymbiosis
Genetic transformation
Kleptoplasty
Kleptoplasty
Dinoflagellates
Genetic transformation
Diatoms
Cryopreservation
Endosymbiosis
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Summary:Endosymbiosis is a widespread and ecologically significant phenomenon in the marine environment. How these endosymbiotic partners evolve into an organism with a new organelle is still mostly unknown and requires investigation into modern symbioses. Dinotoms, dinoflagellates with evolutionarily intermediate diatom plastids, are considered excellent models for studying organellogenesis as they remain at three successive but distinct stages. Efforts to understand the host dinoflagellate-endosymbiotic diatom relationship has been limited by the lack of genetic transformation methods for either member of the symbiosis. To address this absence, we modified existing diatom biolistic and conjugation transformation methods and cryopreservation protocols for the diatom Nitzschia captiva, an essential prey for the kleptoplastic dinotom Durinskia capensis. Through the use of Phaeodactylum tricornutum, Cylindrotheca fusiformis, and native Nitzschia captiva diatom designed plasmids, we successfully express and target EGFP to the cytosol, mitochondria, and plastids of N. captiva, and visualize these organelles inside D. capensis in vivo, allowing specific labeling and tracking of organelles and proteins after ingestion. Furthermore, we attempt to utilize CRISPR/Cas9 to target the introduced EGFP gene but find no evidence of successful gene editing. •Nitzschia captiva is an essential prey diatom for a kleptoplastic dinotom.•We report transformation methods and a cryopreservation protocol for Nitzschia captiva.•EGFP labeled diatom organelles were visualized inside dinoflagellate host in vivo.•These protocols are expected to advance visualization and understanding of dinotoms.
ISSN:2211-9264
2211-9264
DOI:10.1016/j.algal.2023.103136