Citation
Ravigadevi Sambanthamurthi, . and Ghulam Kadir Ahmad Parveez, . and Prufer Dirk, . and Mat Yunus Abdul Masani, . and Noll Gundula A., . (2017) Protoplast transformation system as a potential platform for oil palm genetic engineering. [Proceedings Paper]
Abstract
Protoplasts are a valuable starting material for oil palm genetic engineering because they are totipotent and chimeras can be avoided by regenerating transgenic plants from a single transformed cell. Here were report on oil palm protoplast isolation and plant regeneration as well as protoplast transformation using polyethylene glycol PEG-mediated transfection and DNA microinjection. We observed efficient protoplast isolation from suspension cells digested with enzyme solution comprising cellulase pectinase pectolyase potassium chloride calcium chloride and mannitol. We then validated the optimal factors that influenced the plant regeneration from oil palm protoplasts which depends strongly on the medium composition plant growth regulator PGR and cultivation procedure. For protoplast transformation we found that concentrations of DNA PEG and magnesium chloride and application of heat shock treatment were the important determinants of efficient PEG-mediated transfection. Whereas protoplast embedded in alginate layer cultured for three days and injected with 100 ng/ul DNA solution were the optimal factors for microinjection that enabling us to successfully regenerate transgenic microcalli expressing a green fluorescent protein GFP as a visible marker.
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Abstract
Protoplasts are a valuable starting material for oil palm genetic engineering because they are totipotent and chimeras can be avoided by regenerating transgenic plants from a single transformed cell. Here were report on oil palm protoplast isolation and plant regeneration as well as protoplast transformation using polyethylene glycol PEG-mediated transfection and DNA microinjection. We observed efficient protoplast isolation from suspension cells digested with enzyme solution comprising cellulase pectinase pectolyase potassium chloride calcium chloride and mannitol. We then validated the optimal factors that influenced the plant regeneration from oil palm protoplasts which depends strongly on the medium composition plant growth regulator PGR and cultivation procedure. For protoplast transformation we found that concentrations of DNA PEG and magnesium chloride and application of heat shock treatment were the important determinants of efficient PEG-mediated transfection. Whereas protoplast embedded in alginate layer cultured for three days and injected with 100 ng/ul DNA solution were the optimal factors for microinjection that enabling us to successfully regenerate transgenic microcalli expressing a green fluorescent protein GFP as a visible marker.
Additional Metadata
| Item Type: | Proceedings Paper |
|---|---|
| Additional Information: | Available at Perpustakaan Sultan Abdul Samad Universiti Putra Malaysia 43400 UPM Serdang Selangor Malaysia. SB299 P3I61 2017 Call Number. |
| AGROVOC Term: | Oil palm |
| AGROVOC Term: | Genetic engineering |
| AGROVOC Term: | Genetic transformation |
| AGROVOC Term: | Isolation of microorganisms |
| AGROVOC Term: | Protoplast culture |
| AGROVOC Term: | Suspension culture |
| AGROVOC Term: | Plant growth regulators |
| AGROVOC Term: | Transgenic plants |
| AGROVOC Term: | Protoplasts |
| Geographical Term: | MALAYSIA |
| Depositing User: | Ms. Suzila Mohamad Kasim |
| Last Modified: | 24 Apr 2025 05:15 |
| URI: | http://webagris.upm.edu.my/id/eprint/13116 |
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