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Related Concept Videos

Plant Tissue Culture02:57

Plant Tissue Culture

Plant tissue culture is widely used in both primary and applied science. Applications range from plant development studies to functional gene studies, crop improvement, commercial micropropagation, virus elimination, and conservation of rare species.
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CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

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An efficient protoplast-based genome editing protocol for Vitis species.

David M Tricoli1, Juan M Debernardi1

  • 1Plant Transformation Facility, University of California, Davis, CA 95616, USA.

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|June 19, 2024
PubMed
Summary

Researchers developed an efficient grape protoplast culture method for plant regeneration. This breakthrough enables CRISPR genome editing in various grape varieties, advancing agricultural applications.

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Area of Science:

  • Plant Biotechnology
  • Agricultural Science
  • Genomics

Background:

  • CRISPR-Cas technologies offer precise plant genome modification for agricultural advancement.
  • Efficient delivery and regeneration of edited cells are crucial for CRISPR applications in plants.
  • Protoplast culture is a key method for generating non-chimeric, transgene-free genome-edited plants, especially in vegetatively propagated species like grape.

Purpose of the Study:

  • To establish an efficient protocol for regenerating whole plants from grape protoplasts.
  • To enable CRISPR-Cas genome editing in various *Vitis* species using protoplast culture.
  • To overcome regeneration limitations hindering genome editing in grape.

Main Methods:

  • Grape protoplasts were encapsulated in calcium alginate beads.
  • Protoplasts were co-cultured with feeder cells to promote division and callus formation.
  • Genome editing was performed using CRISPR-plasmid or ribonucleoprotein (RNP) complexes delivered to protoplasts.

Main Results:

  • An efficient plant regeneration protocol from protoplasts was developed for multiple grape varieties (*Vitis vinifera*, rootstocks, *Vitis arizonica*).
  • Regenerated plants exhibited successful genome editing, demonstrated by albino phenotypes resulting from edits in the *VvPHYTOENE DESATURASE* gene.
  • The protocol facilitated transgene-free genome editing in diverse *Vitis* species.

Conclusions:

  • The developed protoplast culture and regeneration protocol is effective for diverse *Vitis* species.
  • This platform significantly enhances the potential for CRISPR-Cas genome editing in grape.
  • The findings pave the way for accelerated crop improvement in grapevines.