ArticlePlant methods2025
PVP-40 mediated enhancement of mesophyll protoplast yield and viability for transient gene expression in black huckleberry.
Article in Plant methods, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 4 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
4 citing papers in PubMed.
- Genome Editing Approaches in Flax (International journal of molecular sciences · 2026Review
- Establishment of a protoplast isolation and transient transformation system for tung tree (Vernicia fordii).Plant methods · 2026Article
- Correction: PVP-40 mediated enhancement of mesophyll protoplast yield and viability for transient gene expression in black huckleberry.Plant methods · 2026Article
- Hyperhydricity Syndrome in In Vitro Plants: Mechanisms, Physiology, and Control.Plants (Basel, Switzerland) · 2025Review
Corrections and comments
- Erratum issued
Authors and funding
3 authors.
Funding
Abstract
backgroundBlack huckleberry (Vaccinium membranaceum) is a native fruit species of high nutritional, medicinal, ecological, and economic value. The black huckleberries, abundant in bioactive compounds, offer significant antioxidants and anti-inflammatory effects and play a key role in maintaining wildlife and forest ecosystems. Despite its importance, protoplast isolation and gene editing have not been reported in this species. These techniques are essential for functional genomics and crop improvement, but the recalcitrant nature of this species, complex genome, and variable ploidy present significant challenges for cellular and molecular manipulation. This study aimed to establish a reliable protocol for efficient mesophyll protoplast isolation and transient gene expression in V. membranaceum using in vitro-grown leaves.
resultsA systematic optimization of enzyme composition, osmotic concentration, antioxidant supplementation, and pH was undertaken to enhance protoplast yield and viability in V. membranaceum. The optimized enzymatic combination of 2% cellulase R-10, 1% hemicellulase, 1% Macerozyme R-10, and 1.5% pectinase facilitated efficient cell wall degradation while maintaining structural integrity. The inclusion of 0.6 M mannitol ensured osmotic stability, and 1% PVP-40 effectively suppressed phenolic oxidation, significantly improving protoplast viability. A near-neutral pH of 5.8 supported optimal enzyme activity without inducing cellular damage. Under these optimized conditions, 14 h enzymatic digestion produced 7.20 × 10⁶ protoplasts g⁻
conclusionsThis study presents a standardized, PVP-40-enhanced protocol for mesophyll protoplast isolation with notable yield and viability in V. membranaceum, supporting efficient transient gene expression. The method provides a robust platform for functional genomics, gene editing, and biotechnological applications in this underutilized species and other related plant species.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.