Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.)

Daniel Grogg, Marius Rohner, Steven Yates, Chloe Manzanares, Simon E. Bull, Sue Dalton, Maurice Bosch, Bruno Studer, Giovanni A.L. Broggini*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Citations (SciVal)
51 Downloads (Pure)

Abstract

Genetic transformation of perennial ryegrass (Lolium perenne L.) is critical for fundamental and translational research in this important grass species. It often relies on Agrobacterium-mediated transformation of callus tissue. However, callus induction is restricted to a few genotypes that respond well to tissue culture. Here, we report callus induction from different perennial ryegrass genotypes and explants, such as shoot tips, seeds, and anthers, which were transformed with several plasmids for functional genomics. β-glucuronidase (GUS) histochemical staining showed the LmdsRNAbp promoter sequence was active in stigmas, spikelets, anthers, and leaves. We also transformed calli with plasmids allowing gene silencing and gene knock-out using RNA interference and CRISPR/Cas9, respectively, for which genotypic and phenotypic investigations are ongoing. Using 19 different constructs, 262 transgenic events were regenerated. Moreover, the protocol regenerated a doubled haploid transgenic event from anther-derived calli. This work provides a proof-of-concept method for expanding the range of genotypes amenable to transformation, thus, serving research and breeding initiatives to improve this important grass crop for forage and recreation.

Original languageEnglish
Article numbere2054
Number of pages19
JournalPlants
Volume11
Issue number15
DOIs
Publication statusPublished - 05 Aug 2022

Keywords

  • Agrobacterium-mediated transformation
  • doubled haploid (DH)
  • functional genomics
  • genome editing
  • perennial ryegrass (Lolium perenne L.)
  • tissue culture

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