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

doi:10.3390/plants13071044

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

Department of Agroecology - Crop Genetics and Biotechnology

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

3 Citations (Scopus)

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 language	English
Article number	1044
Journal	Plants
Volume	13
Issue	7
Number of pages	11
ISSN	2223-7747
DOIs	https://doi.org/10.3390/plants13071044
Publication status	Published - Apr 2024

Keywords

de novo domestication
protoplasts
genome editing
wild potato
transgene free

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10.3390/plants13071044Licence: CC BY

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title = "A Walk on the Wild Side:: Genome Editing of Tuber-Bearing Solanum bulbocastanum",

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.",

keywords = "de novo domestication, protoplasts, genome editing, wild potato, transgene free",

author = "Aristotelis Azariadis and Andrzejczak, {Olga Agata} and Carlsen, {Frida M.} and Ida Westberg and Henrik Brinch-Pedersen and Pedersen, {Bent L.} and Kim Hebelstrup",

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month = apr,

doi = "10.3390/plants13071044",

language = "English",

volume = "13",

journal = "Plants",

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T2 - Genome Editing of Tuber-Bearing Solanum bulbocastanum

AU - Azariadis, Aristotelis

AU - Andrzejczak, Olga Agata

AU - Carlsen, Frida M.

AU - Westberg, Ida

AU - Brinch-Pedersen, Henrik

AU - Pedersen, Bent L.

AU - Hebelstrup, Kim

PY - 2024/4

Y1 - 2024/4

N2 - 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.

AB - 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.

KW - de novo domestication

KW - protoplasts

KW - genome editing

KW - wild potato

KW - transgene free

UR - http://www.scopus.com/inward/record.url?scp=85190124991&partnerID=8YFLogxK

U2 - 10.3390/plants13071044

DO - 10.3390/plants13071044

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SN - 2223-7747

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JO - Plants

JF - Plants

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M1 - 1044

ER -

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

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Keywords

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