Transformation of the dinoflagellate chloroplast
2019
Dinoflagellatealgae are ecologically and environmentally important, as symbionts of corals and many other aquatic organisms, and the causative agents of
red tides. However, attempts over the last twenty years to establish genetic manipulation systems for
dinoflagellateshave met with little success. We have exploited the unusual
chloroplastgenome of
dinoflagellatesto establish a system for transformation of this organelle. The
chloroplastgenome of
peridinin-containing (the ancestral state)
dinoflagellatesis highly reduced and composed of a number of small, plasmid-like molecules, referred to as ‘
minicircles’. We have constructed
shuttle vectorsthat are fusions of
minicirclesand Escherichia coli plasmids and carry
selectable markers. We used
biolistictransformation to introduce these into the model
dinoflagellate
Amphidinium carterae. We found that the plasmids confer the expected phenotype on the
dinoflagellatecells, and we can detect the plasmid DNA and associated transcripts following selection, indicating successful transformation. This opens up the possibility of studying many aspects of
dinoflagellate
chloroplastbiology, including the maintenance and expression of the
minicircles, and the role of the
chloroplastin phenomena such as
coral bleaching.
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