An efficient woody plant protoplast platform enables transgene-free multiplex genome editing and rapid trait validation in pear

Clicks: 1
ID: 316110
2026
Free PDF
Article Quality & Performance Metrics
Overall Quality
Not rated
Combines reader engagement with the AI quality analysis. This article has not been analysed, so there is no overall score — reader engagement is measured and shown alongside.
AI Quality Assessment
Not analyzed
Readership in this journal

Ranked #235 of 280 articles by views in Journal of experimental botany

Most read Least read

Bar heights use a square-root scale. Only the 120 most-read articles are drawn; the journal has 280 in total.

Mint this article as an NFT
Not yet minted

Create a permanent, verifiable on-chain record of this article on the Scimatic Network. The NFT is held in your Journament account, and you can withdraw it to your own wallet at any time.

5 SUSD one-off · no wallet required
Sign in to mint
Abstract
Multiplex editing is crucial for analyzing complex multiple-gene traits in woody plants, yet its application remains limited by low transformation efficiency and lengthy regeneration cycles. To overcome these barriers, this study establishes an efficient protoplast isolation protocol for Pyrus, employing 1.0% cellulase R10 and 0.4% macerozyme R10 with an 8.5 h digestion, and demonstrates its broad applicability across seven economically important woody plants. Coupling a 40% PEG-4000-mediated transfection regimen with DNA-free CRISPR/Cas9 ribonucleoprotein (RNP) delivery enabled multiplex genome editing in isolated protoplasts. Using this platform, simultaneous disruption of PbrARC3, PbrPARC6, and PbrFtsZ2-1a, key components of the chloroplast division apparatus, consistently reproduced macro-chloroplast abnormalities, confirming effective multigene perturbation within a single cellular context. Notably, chloroplast division failure activated chloroplast-to-nucleus retrograde signaling, evidenced by induction of nuclear stress-response genes PbrRBOHD and PbrZAT12, a concomitant surge in reactive oxygen species, and progression to severe cellular deformation. These results establish a rapid, cross-genus protoplast-RNP workflow that enables DNA-free multiplex editing and accelerates genotype-to-phenotype analyses in woody perennials. The approach provides a practical foundation for functional genomics and supports advances in non-transgenic precision breeding of tree crops.
Reference Key
openalex_W7163734479 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Yun Shao, C J Li, Yanni He, Zhihua Xie, Kaijie Qi, Chao Gu, Shaoling Zhang
Journal Journal of experimental botany
Year 2026
DOI
10.1093/jxb/erag282
URL
https://doi.org/10.1093/jxb/erag282
Keywords Keywords not found

Citations

No citations found. To add a citation, contact the admin at info@scimatic.org

Comments

No comments yet. Be the first to comment on this article.