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Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.)
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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9370183/ https://www.ncbi.nlm.nih.gov/pubmed/35956532 http://dx.doi.org/10.3390/plants11152054 |
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author | Grogg, Daniel Rohner, Marius Yates, Steven Manzanares, Chloe Bull, Simon E. Dalton, Sue Bosch, Maurice Studer, Bruno Broggini, Giovanni A. L. |
author_facet | Grogg, Daniel Rohner, Marius Yates, Steven Manzanares, Chloe Bull, Simon E. Dalton, Sue Bosch, Maurice Studer, Bruno Broggini, Giovanni A. L. |
author_sort | Grogg, Daniel |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-9370183 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-93701832022-08-12 Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.) Grogg, Daniel Rohner, Marius Yates, Steven Manzanares, Chloe Bull, Simon E. Dalton, Sue Bosch, Maurice Studer, Bruno Broggini, Giovanni A. L. Plants (Basel) Article 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. MDPI 2022-08-05 /pmc/articles/PMC9370183/ /pubmed/35956532 http://dx.doi.org/10.3390/plants11152054 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Grogg, Daniel Rohner, Marius Yates, Steven Manzanares, Chloe Bull, Simon E. Dalton, Sue Bosch, Maurice Studer, Bruno Broggini, Giovanni A. L. Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.) |
title | Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.) |
title_full | Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.) |
title_fullStr | Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.) |
title_full_unstemmed | Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.) |
title_short | Callus Induction from Diverse Explants and Genotypes Enables Robust Transformation of Perennial Ryegrass (Lolium perenne L.) |
title_sort | callus induction from diverse explants and genotypes enables robust transformation of perennial ryegrass (lolium perenne l.) |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9370183/ https://www.ncbi.nlm.nih.gov/pubmed/35956532 http://dx.doi.org/10.3390/plants11152054 |
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