ArticleInternational journal of molecular sciences2022
A Rapid and Efficient Method for Isolation and Transformation of Cotton Callus Protoplast.
Article in International journal of molecular sciences, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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15 citing papers in PubMed, 38 citations in OpenAlex.
- Optimization of CRISPR/Cas9-mediated CtPDS knockout in guar protoplasts.The plant genome · 2026Article
- A GhBGH2-GhGLK1 Regulatory Module Mediates Salt Tolerance in Cotton.Plant biotechnology journal · 2025Article
- Targeted genome modification in protoplasts of a tea cultivar Kolkhida using RNA-guided Cas9 endonuclease.AoB PLANTS · 2025Article
- An efficient hairy root system for validation of CRISPR/Cas system activities in cotton.Functional & integrative genomics · 2025Article
- Plant genetic transformation: achievements, current status and future prospects.Plant biotechnology journal · 2025Review
- Mediated Transformation of Tamarillo (Plants (Basel, Switzerland) · 2025Article
- A Comparison of DNA-Methylation during Protoplast Culture of Ponkan Mandarin (Plants (Basel, Switzerland) · 2024Article
- Article
- Isolation, Purification, and Application of Protoplasts and Transient Expression Systems in Plants.International journal of molecular sciences · 2023Review
- Establishment of protoplasts transient expression system in Pinellia ternata (Thunb.) Breit.Biotechnology letters · 2023Article
- Using A Protoplast Transformation System to Enable Functional Studies inInternational journal of molecular sciences · 2023Article
- Overexpression of thePlants (Basel, Switzerland) · 2023Article
- Establishment of an efficient cotton root protoplast isolation protocol suitable for single-cell RNA sequencing and transient gene expression analysis.Plant methods · 2023Article
- Genome-wide identification and analyses of cotton high-affinity nitrate transporter 2 family genes and their responses to stress.Frontiers in plant science · 2023Article
- A high-efficiency PEG-CaFrontiers in plant science · 2022Article
Corrections and comments
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Authors and funding
10 authors at 2 institutions in 1 country.
Funding
Abstract
Protoplasts, which lack cell walls, are ideal research materials for genetic engineering. They are commonly employed in fusion (they can be used for more distant somatic cell fusion to obtain somatic hybrids), genetic transformation, plant regeneration, and other applications. Cotton is grown throughout the world and is the most economically important crop globally. It is therefore critical to study successful extraction and transformation efficiency of cotton protoplasts. In the present study, a cotton callus protoplast extraction method was tested to optimize the ratio of enzymes (cellulase, pectinase, macerozyme R-10, and hemicellulase) used in the procedure. The optimized ratio significantly increased the quantity and activity of protoplasts extracted. We showed that when enzyme concentrations of 1.5% cellulase and 1.5% pectinase, and either 1.5% or 0.5% macerozyme and 0.5% hemicellulase were used, one can obtain increasingly stable protoplasts. We successfully obtained fluorescent protoplasts by transiently expressing fluorescent proteins in the isolated protoplasts. The protoplasts were determined to be suitable for use in further experimental studies. We also studied the influence of plasmid concentration and transformation time on protoplast transformation efficiency. When the plasmid concentration reaches 16 µg and the transformation time is controlled within 12-16 h, the best transformation efficiency can be obtained. In summary, this study presents efficient extraction and transformation techniques for cotton protoplasts.
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