| Autor: |
B. Deng, X. Fu, B. Peng, Q. Miao, S. Zeng, K. Tang, Q. Pan |
| Jazyk: |
angličtina |
| Rok vydání: |
2023 |
| Předmět: |
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| Zdroj: |
Biologia Plantarum, Vol 67, Iss 1, Pp 142-149 (2023) |
| Druh dokumentu: |
article |
| ISSN: |
1573-8264 |
| DOI: |
10.32615/bp.2023.012 |
| Popis: |
Artemisia japonica Thunb is an important perennial herb containing abundant chemical compounds utilized in conventional medicine for the treatment of malaria, hepatitis, hypertension, and inflammation. The protoplasts-based transient transformation system is a versatile and convenient tool for functional gene analysis in several Artemisia species. However, effective protoplast preparation and transformation systems are still lacking for A. japonica. We developed an efficient protoplast-based transformation system by optimizing conditions of protoplasts isolation and polyethylene glycol (PEG)-mediated transformation in A. japonica. The optimum conditions for the protoplast preparation were: the enzyme solution containing 1.75% (m/v) cellulase R10, 0.5% (m/v) macerozyme R-10, and 0.4 M D-mannitol with proper leaves treatment and pre-plasmolysis treatment. The maximum protoplast yield was 1.93 × 106 protoplasts g-1(FM) and the viability of protoplasts was approximately 87.5% under optimized conditions using an orthogonal experiment. Furthermore, the transient protoplast transformation efficiency was 47.86% in A. japonica protoplast under the conditions of 40% (m/v) PEG 4000 for 20 min. The establishment of A. japonica protoplasts isolation and transient transformation system can accelerate the gene function studies of A. japonica and provide a fast and simple gene expression platform for molecular, biochemical, and functional gene characterization for other Artemisia species. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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