First-generation genome editing in potato using hairy root transformation

Nathaniel M. Butler, Shelley H. Jansky, Jiming Jiang

Research output: Contribution to journalArticlepeer-review

36 Scopus citations

Abstract

Genome editing and cis-gene breeding have rapidly accelerated crop improvement efforts, but their impacts are limited by the number of species capable of being genetically transformed. Many dicot species, including some vital potato relatives being used to accelerate breeding and genetics efforts, remain recalcitrant to standard Agrobacterium tumefaciens-based transformation. Hairy root transformation using Agrobacterium rhizogenes (A. rhizogenes) provides an accelerated approach to generating transgenic material but has been limited to analysis of hairy root clones. In this study, strains of A. rhizogenes were tested in the wild diploid potato relative Solanum chacoense, which is recalcitrant to infection by Agrobacterium tumefaciens. 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 and regenerated. We found that 64%–98% of transgenic hairy root clones expressing CRISPR/Cas9 reagents carried targeted mutations, while only 14%–30% of mutations were chimeric. The mutations were maintained in regenerated lines as stable mutations at rates averaging at 38% and were capable of germ-line transmission to progeny. This novel approach broadens the numbers of genotypes amenable to Agrobacterium-mediated transformation while reducing chimerism in primary events and accelerating the generation of edited materials.

Original languageEnglish (US)
Pages (from-to)2201-2209
Number of pages9
JournalPlant Biotechnology Journal
Volume18
Issue number11
DOIs
StatePublished - Nov 1 2020
Externally publishedYes

Bibliographical note

Funding Information:
and strains were graciously provided by the laboratories of Dan Voytas (University of Minnesota – Twin Cities) and Jean‐Michel Ané (University of Wisconsin – Madison), respectively. This project was supported by the Postdoctoral Research Fellowships in Biology Program award no. IOS‐1523876 from the NSF National Plant Genome Initiative. Agrobacterium tumefaciens A. rhizogenese

Funding Information:
Agrobacterium tumefaciens and A. rhizogenese strains were graciously provided by the laboratories of Dan Voytas (University of Minnesota ? Twin Cities) and Jean-Michel An? (University of Wisconsin ? Madison), respectively. This project was supported by the Postdoctoral Research Fellowships in Biology Program award no. IOS-1523876 from the NSF National Plant Genome Initiative.

Publisher Copyright:
© 2020 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

Keywords

  • Agrobacterium rhizogenes
  • Cas
  • CRISPR
  • crop
  • Csy4
  • gene editing
  • germ-line
  • plant transformation
  • reagent delivery
  • targeted mutagenesis
  • Trex2

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