Cloning and Functional Characterization of Chalcone Isomerase Genes Involved in Anthocyanin Biosynthesis in <i>Clivia miniata</i>
Bibliographic record
Abstract
Chalcone isomerase (CHI), catalyzing isomerization of chalcones, is a crucial enzyme in flavonoid biosynthesis. Three CHI genes were isolated from Clivia miniata and designated as CmCHI1, CmCHI2 and CmCHI3, respectively. Multiple sequence alignments and phylogenetic analysis showed that CmCHI1 and CmCHI2 were members of type I CHI proteins, whereas CmCHI3 belonged to type IV CHI proteins. Subcellular localization analysis found that all three CmCHIs had diffused distribution in the cytoplasm similar to green fluorescent protein (GFP). Anthocyanin biosynthesis and gene expression analysis demonstrated that CmCHIs were highly expressed in anthocyanin accumulated tissues. To further functionally characterize the role of CmCHIs, an in vitro enzymatic activity assay was carried out using the purified recombinant proteins. Results showed that CmCHI1 and CmCHI2 could completely convert the substrate naringenin chalcone (NC) into the product naringenin (NA), whereas CmCHI3 seemed nonfunctional as no increment of NA was detected. Further genetic transformation of Arabidopsis tt5-1 mutant validated that CmCHI1 and CmCHI2 rather than CmCHI3 could complement the chi deficient phenotypes. In summary, CmCHI1 and CmCHI2 are the real active CHI genes in Clivia miniata. The results not only broaden our knowledge on flavonoid biosynthesis in C. miniata but also lay a new foundation for further flavonoid modification in C. miniata.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.001 |
| 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 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 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 source (direct Gemma or distilled Codex), 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".