A versatile protoplast system and its application in <i>Cannabis sativa</i> L.
Bibliographic record
Abstract
Cannabis sativa L. (cannabis) is one of the earliest domesticated crops cultivated for medicinal, food, industrial, and recreational use throughout the world. A growing number of jurisdictions have lifted the prohibition on medical and recreational cannabis with legislation that allows researchers to conduct important and urgently needed research, such as understanding the molecular basis of agronomically important traits. However, a dearth of suitable molecular genetic techniques for the analysis of cannabis species has hampered the elucidation of gene function. Here, we developed a robust protoplast transient transformation system by optimizing conditions of protoplast isolation and polyethylene glycol–mediated transformation using cannabis cotyledons. A protoplast isolation yield of up to 1.15 × 10 7 cells per gram of fresh material and 98.5% viability and a transformation efficiency of 75.4% were achieved. This optimized protocol for protoplast isolation and transformation was successfully applied to eight representative cannabis varieties, which highlight the versatility and robustness of this technique. Using this protocol, the nuclear localization of a basic helix–loop–helix transcription factor (CsMYC2) was visualized, and the transcriptional activity of CsMYC2 was assessed. These results demonstrate the effectiveness of our cannabis protoplast isolation and transient transformation method that should serve as a powerful molecular research tool in a wide range of molecular studies.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".