A Walk on the Wild Side: Genome Editing of Tuber-Bearing Solanum bulbocastanum

Aristotelis Azariadis, Olga Agata Andrzejczak*, Frida M. Carlsen, Ida Westberg, Henrik Brinch-Pedersen, Bent L. Pedersen, Kim Hebelstrup*

*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

Solanum bulbocastanum is a wild diploid tuber-bearing plant. We here demonstrate transgene-free genome editing of S. bulbocastanum protoplasts and regeneration of gene-edited plants. We use ribonucleoproteins, consisting of Cas9 and sgRNA, assembled in vitro, to target a gene belonging to the nitrate and peptide transporter family. Four different sgRNAs were designed and we observed efficiency in gene-editing in the protoplast pool between 8.5% and 12.4%. Twenty-one plants were re-generated from microcalli developed from individual protoplasts. In three of the plants we found that the target gene had been edited. Two of the edited plants had deletion mutations introduced into both alleles, whereas one only had a mutation in one of the alleles. Our work demonstrates that protocols for the transformation of Solanum tuberosum can be optimized to be applied to a wild Solanum species.
Original languageEnglish
Article number1044
JournalPlants
Volume13
Issue7
Number of pages11
ISSN2223-7747
DOIs
Publication statusPublished - Apr 2024

Keywords

  • de novo domestication
  • protoplasts
  • genome editing
  • wild potato
  • transgene free

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