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Next-generation marker-free transplastomic plants: engineering the chloroplast genome without integration of marker genes in Solanum tuberosum (potato). | LitMetric

AI Article Synopsis

  • - This study presents an improved method for creating marker-free transgenic plants by engineering their plastids (the part of the cell involved in photosynthesis and other functions) to integrate new genes without leaving behind antibiotic selection markers.
  • - The technique was applied to potato plants, successfully achieving full integration of the desired transgenes at homoplasmy (uniformity in genetic makeup) after just one round of tissue culture.
  • - The resulting plants displayed normal traits and stable traits over generations, suggesting this new approach could facilitate the commercial use of transplastomic plants in agriculture.

Article Abstract

This study describes an optimized plastid genetic engineering platform to produce full marker-free transplastomic plants with transgene integrated at homoplasmy in one step in tissue culture. Plastid engineering is attractive for both biotechnology and crop improvement due to natural bio-confinement from maternal inheritance, the absence of transgene positional effects and silencing, the ability to express transgenes in operons, and unparalleled production of heterologous proteins. While plastid engineering has had numerous successes in the production of high-value compounds, no transplastomic plants have been approved for use in agriculture. In order for transplastomic plants to be used in agriculture, the removal of antibiotic selection genes is required. In this work, we developed an optimized strategy to generate homoplasmic marker-free lines of potato (Solanum tuberosum) in a single transformation event. To achieve marker-free transplastomic lines, vectors were redesigned to enable integration of the transgene cassette into the plastid genome, while maintaining the selection cassette on the vector backbone. After an initial round of tissue culture with selection, the selective pressure was removed, leading to the elimination of the vector backbone, while retaining the integrated transgene cassette at homoplasmy. Marker-free transplastomic lines produced using this strategy had a normal phenotype, and transgene integration was stable across generations. The new vectors developed in this work for the generation of marker-free transplastomics will represent a valuable alternative platform for routine plastid genetic engineering in higher plants. It is also anticipated that this approach will contribute to speed the path to commercialization of these novel transplastomic plant varieties.

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Source
http://dx.doi.org/10.1007/s00299-024-03375-9DOI Listing

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