Journal
FRONTIERS IN PLANT SCIENCE
Volume 9, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fpls.2018.00439
Keywords
Nannochloropsis; plastid transformation; oleaginous microalga; green fluorescent protein; photosynthetic cell factory
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Funding
- Natural Science Foundation of Hainan Province [317010]
- Project of State Key Laboratory of Marine Resource Utilization in South China Sea [2018004, 2016005]
- Foundation of Hainan University [KYQD1561]
- Project of Innovation & Development of Marine Economy [2017-285]
- National Natural Science Foundation of China [31401705, 31660744, 41466002]
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Plastid engineering offers an important tool to fill the gap between the technical and the enormous potential of microalgal photosynthetic cell factory. However, to date, few reports on plastid engineering in industrial microalgae have been documented. This is largely due to the small cell sizes and complex cell-wall structures which make these species intractable to current plastid transformation methods (i.e., biolistic transformation and polyethylene glycol-mediated transformation). Here, employing the industrial oleaginous microalga Nannochloropsis oceanica as a model, an electroporation-mediated chloroplast transformation approach was established. Fluorescent microscopy and laser confocal scanning microscopy confirmed the expression of the green fluorescence protein, driven by the endogenous plastid promoter and terminator. Zeocin-resistance selection led to an acquisition of homoplasmic strains of which a stable and site-specific recombination within the chloroplast genome was revealed by sequencing and DNA gel blotting. This demonstration of electroporation-mediated chloroplast transformation opens many doors for plastid genome editing in industrial microalgae, particularly species of which the chloroplasts are recalcitrant to chemical and microparticle bombardment transformation.
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