ArticleBMC biotechnology2017
Highly efficient mesophyll protoplast isolation and PEG-mediated transient gene expression for rapid and large-scale gene characterization in cassava (Manihot esculenta Crantz).
Article in BMC biotechnology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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23 citing papers in PubMed, 74 citations in OpenAlex.
- Metabolic Enzyme MeHNL11 Regulates MeCAS1b Transcription for Cyanide Reutilization in Response to Nitrate Deficiency in Cassava.Plant biotechnology journal · 2026Article
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- Overexpression ofInternational journal of molecular sciences · 2024Article
- A high-efficient protoplast transient system for screening gene editing elements in Salvia miltiorrhiza.Plant cell reports · 2024Article
- Isolation, Purification, and Application of Protoplasts and Transient Expression Systems in Plants.International journal of molecular sciences · 2023Review
- An efficient method for protoplast-mediated production of transformed castor bean (Ricinus communis) lines.BMC research notes · 2023Article
- Optimization of Isolation and Transformation of Protoplasts fromInternational journal of molecular sciences · 2023Article
- Cotyledon peeling method for passion fruit protoplasts: a versatile cell system for transient gene expression in passion fruitFrontiers in plant science · 2023Article
- CRISPR/Cas9: an advanced platform for root and tuber crops improvement.Frontiers in genome editing · 2023Review
- Isolation, purification and PEG-mediated transient expression of mesophyll protoplasts in Camellia oleifera.Plant methods · 2022Article
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- Plant regeneration from leaf mesophyll derived protoplasts of cassava (Manihot esculenta Crantz).PloS one · 2022Article
- Optimization of Protoplast Isolation from Leaf Mesophylls of Chinese Cabbage (Plants (Basel, Switzerland) · 2021Article
- Efficient isolation and purification of tissue-specific protoplasts from tea plants (Camellia sinensis (L.) O. Kuntze).Plant methods · 2021Article
- Highly Efficient Leaf Base Protoplast Isolation and Transient Expression Systems for Orchids and Other Important Monocot Crops.Frontiers in plant science · 2021Article
- A cassava protoplast system for screening genes associated with the response to South African cassava mosaic virus.Virology journal · 2020Article
- Highly Efficient Protoplast Isolation and Transient Expression System for Functional Characterization of Flowering Related Genes inInternational journal of molecular sciences · 2020Article
- Development and validation of a novel and robust cell culture system in soybean (Glycine max (L.) Merr.) for promoter screening.Plant cell reports · 2019Article
- Isolation of Powdery Mildew Haustoria from Infected Barley.Bio-protocol · 2019Article
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundCassava (Manihot esculenta Crantz) is a major crop extensively cultivated in the tropics as both an important source of calories and a promising source for biofuel production. Although stable gene expression have been used for transgenic breeding and gene function study, a quick, easy and large-scale transformation platform has been in urgent need for gene functional characterization, especially after the cassava full genome was sequenced.
methodsFully expanded leaves from in vitro plantlets of Manihot esculenta were used to optimize the concentrations of cellulase R-10 and macerozyme R-10 for obtaining protoplasts with the highest yield and viability. Then, the optimum conditions (PEG4000 concentration and transfection time) were determined for cassava protoplast transient gene expression. In addition, the reliability of the established protocol was confirmed for subcellular protein localization.
resultsIn this work we optimized the main influencing factors and developed an efficient mesophyll protoplast isolation and PEG-mediated transient gene expression in cassava. The suitable enzyme digestion system was established with the combination of 1.6% cellulase R-10 and 0.8% macerozyme R-10 for 16 h of digestion in the dark at 25 °C, resulting in the high yield (4.4 × 10
conclusionWe optimized the main influencing factors and developed an efficient mesophyll protoplast isolation and transient gene expression in cassava, which will facilitate large-scale characterization of genes and pathways in cassava.
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