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Protocols for Grapevine Transformation: Tissue Preparation, Protoplast Isolation, and Cultivation.

Rebeka Strah1, Edoardo Bertini2, Sara Zenoni2

  • 1Department of Biotechnology and Systems Biology, National institute of Biology, Ljubljana, Slovenia. Rebeka.strah@nib.si.

Methods in Molecular Biology (Clifton, N.J.)
|January 13, 2026
PubMed
Summary
This summary is machine-generated.

Developing effective grapevine genetic transformation protocols is crucial for studying phytoplasma diseases. This research provides reproducible methods for grapevine tissue culture and protoplast isolation, enabling direct host-plant interaction studies.

Keywords:
CallusEmbryogenicGrapevineProtoplastTransfectionTransformation

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Area of Science:

  • Plant pathology
  • Plant biotechnology
  • Molecular biology

Background:

  • Grapevine (Vitis vinifera) is economically significant but vulnerable to phytoplasma infections.
  • Current research on plant-phytoplasma interactions often uses non-host species due to difficulties in grapevine genetic manipulation.
  • Studying interactions directly in grapevine hosts is essential for accurate disease mechanism insights.

Purpose of the Study:

  • To establish reliable protocols for grapevine genetic transformation.
  • To enable direct study of plant-phytoplasma interactions within the host plant.
  • To provide reproducible methods for grapevine tissue culture and protoplast isolation.

Main Methods:

  • Collection and preparation of various grapevine tissues for transformation.
  • Optimization of protoplast isolation from grapevine cells.
  • Cultivation and regeneration protocols for grapevine protoplasts.

Main Results:

  • Successfully established protocols for generating transformation-competent grapevine material.
  • Developed efficient procedures for isolating and culturing grapevine protoplasts.
  • Demonstrated the feasibility of using these methods for future host-pathogen interaction studies.

Conclusions:

  • The presented protocols overcome existing limitations in grapevine genetic manipulation.
  • This work facilitates direct investigation of phytoplasma-grapevine interactions.
  • The methods are crucial for advancing our understanding of grapevine phytoplasma diseases.