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ARS Home » Midwest Area » Madison, Wisconsin » Vegetable Crops Research » Research » Publications at this Location » Publication #367447
Research Project: Trait Discovery, Genetics, and Enhancement of Allium, Cucumis, and Daucus Germplasm

Location: Vegetable Crops Research

Title: First generation genome editing in potato using hairy root transformation

Author
item Butler, Nathaniel
item Jansky, Shelley
item JIANG, JIMING - Michigan State University

Submitted to: Plant Biotechnology Journal
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/23/2020
Publication Date: 3/14/2020
Citation: Butler, N.M., Jansky, S.H., Jiang, J. 2020. First generation genome editing in potato using hairy root transformation. Plant Biotechnology Journal. p.1-9. https://doi.org/10.1111/pbi.13376.
DOI: https://doi.org/10.1111/pbi.13376

Interpretive Summary: Genome editing using technologies such as CRISPR/Cas9 has opened the door to accelerate crop improvement beyond what has been possible with conventional GMO technologies. However, not all crop species can be genetically modified, limiting the number of crop species that can benefit from these exciting new technologies. This study aimed to broaden the number of crop species capable of being genetically modified by testing a difference species of Agrobacterium that is used to deliver DNA to plant cells. This species of Agrobacterium, Agrobacterium rhizogenes was capable of genetically modifying an important potato line previously unable to be genetically modified. Furthermore, the genome editing modifications could be generated more quickly, saving time and accelerating the breeding process. This approach can be used in crop species that are difficult to genetically transform and accelerate the genome editing process.

Technical Abstract: Genome editing and cis-gene breeding have rapidly accelerated plant genetic engineering but are limited by the number of crops species capable of being genetically transformed. Recent advancements in monocot transformation have greatly broadened the number of crop species capable of being transformed but many dicot species remain recalcitrant to standard transformation approaches. In this study, strains of Agrobacterium rhizogenes (A. rhizogenes) were tested against a diploid, self-compatible potato recalcitrant to standard strains of Agrobacterium tumefaciens (LBA4404 and GV3101). One strain of A. rhizogenes, MSU440 emerged as being capable of delivering a T-DNA carrying the GUS marker and generating transgenic hairy root clones capable of GUS expression and regeneration to whole plants. CRISPR/Cas9 reagents targeting the potato PHYTOENE DESATURASE (StPDS) gene were expressed in hairy root clones to test if targeted mutations could be generated. 64-98% transgenic hairy root clones expressing CRISPR/Cas9 reagents carried targeted mutations, 14-30% of which were chimeric. Targeted mutations were maintained in regenerated lines and were capable of germline transmission. This novel approach broadens the numbers of species compatible with Agrobacterium-mediated transformation while reducing chimerism in primary events and accelerating the generation of edited materials.