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| Content Provider | Springer Nature : BioMed Central |
|---|---|
| Author | Osmani, Zhila Islam, Muhammad Amirul Wang, Feng Meira, Sabrina Rodrigues Kulka, Marianna |
| Abstract | Nanomaterial-mediated plant genetic engineering holds promise for developing new crop cultivars but can be hindered by nanomaterial toxicity to protoplasts. We present a fast, high-throughput method for assessing protoplast viability using resazurin, a non-toxic dye converted to highly fluorescent resorufin during respiration. Protoplasts isolated from hypocotyl canola (Brassica napus L.) were evaluated at varying temperatures (4, 10, 20, 30 ˚C) and time intervals (1–24 h). Optimal conditions for detecting protoplast viability were identified as 20,000 cells incubated with 40 µM resazurin at room temperature for 3 h. The assay was applied to evaluate the cytotoxicity of silver nanospheres, silica nanospheres, cholesteryl-butyrate nanoemulsion, and lipid nanoparticles. The cholesteryl-butyrate nanoemulsion and lipid nanoparticles exhibited toxicity across all tested concentrations (5-500 ng/ml), except at 5 ng/ml. Silver nanospheres were toxic across all tested concentrations (5-500 ng/ml) and sizes (20–100 nm), except for the larger size (100 nm) at 5 ng/ml. Silica nanospheres showed no toxicity at 5 ng/ml across all tested sizes (12–230 nm). Our results highlight that nanoparticle size and concentration significantly impact protoplast toxicity. Overall, the results showed that the resazurin assay is a precise, rapid, and scalable tool for screening nanomaterial cytotoxicity, enabling more accurate evaluations before using nanomaterials in genetic engineering. |
| Related Links | https://plantmethods.biomedcentral.com/counter/pdf/10.1186/s13007-024-01289-x.pdf |
| Ending Page | 14 |
| Page Count | 14 |
| Starting Page | 1 |
| File Format | HTM / HTML |
| ISSN | 17464811 |
| DOI | 10.1186/s13007-024-01289-x |
| Journal | Plant Methods |
| Issue Number | 1 |
| Volume Number | 20 |
| Language | English |
| Publisher | BioMed Central |
| Publisher Date | 2024-10-30 |
| Access Restriction | Open |
| Subject Keyword | Plant Sciences Biological Techniques Fluorescence Absorbance Cell viability Protoplast Resazurin Canola |
| Content Type | Text |
| Resource Type | Article |
| Subject | Plant Science Biotechnology Genetics |
| Journal Impact Factor | 4.7/2023 |
| 5-Year Journal Impact Factor | 5.6/2023 |
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