Heterologous Expression of Phosphite Dehydrogenase in the Chloroplast or Nucleus Enables Phosphite Utilization and Genetic Selection in Picochlorum spp.

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Abstract

Microalgae present a path to ameliorate problems associated with climate change via capture and reduction of CO2 to sustainable fuels and chemicals. Picochlorum is a genus of algae recently recognized for potential application in these regards due to its high productivity, thermotolerance, and halotolerance. Foundational genetic tools have recently been established in this genus. However, at present, genetic markers are limited, hindering genetic throughput and trait stacking approaches. To expand the suite of genetic tools and markers available for this genus, we sought to heterologously express the phosphite dehydrogenase (ptxD) gene from Pseudomonas stutzeri WM88 in both the nucleus and chloroplast of Picochlorum renovo and Picochlorum celeri. The resultant strains allow for utilization of phosphite as a sole phosphorous source and as a nuclear and plastidial selection marker for genetic engineering. Growth analysis indicated comparable growth and composition when transgenic algae were grown in media containing phosphite as a sole phosphorus source, as compared to the conventionally used phosphate. Combined, these results expand the genetic toolbox available to the Picochlorum genus and present a potential crop protection and biocontainment strategy.

Original languageAmerican English
Article number102604
Number of pages6
JournalAlgal Research
Volume62
DOIs
StatePublished - 2022

Bibliographical note

Publisher Copyright:
© 2022 The Authors

NREL Publication Number

  • NREL/JA-2700-80931

Keywords

  • Chloroplast transformation
  • Genetic engineering
  • Microalgae
  • Phosphite
  • Phosphite dehydrogenase
  • Picochlorum

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