A pea antisense gene for the chloroplast stromal processing peptidase yields seedling lethals in <i>Arabidopsis</i>: survivors show defective GFP import <i>in vivo</i>
Bibliographic record
Abstract
The stromal processing peptidase (SPP) of chloroplasts is a metalloendopeptidase that cleaves in vitro a broad range of precursor substrates. Here, we have investigated SPP's role in vivo. Two pea cDNA antisense constructs encoding either full-length SPP (AS4.0) or its N-terminal half (AS2.2) are introduced into Arabidopsis, which contains one gene for SPP that codes for one isoform. Our analyses show that AS4.0 produces a strong mutant phenotype, with a large percentage of the plants dying as seedling lethals. Surviving plants exhibited slower shoot and root growth, and grossly aberrant leaf morphology. Green and white sectoring, and purple pigmentation was observed. In cells where chloroplasts could be identified, they were fewer in number by at least 40%, thylakoids were not fully developed, and starch granules accumulated. The phenotype produced by AS2.2 was less severe. Using green fluorescent protein (GFP) fused to a transit peptide as a reporter, we examined import into chloroplasts in vivo. In the Arabidopsis antisense lines, GFP was located primarily in the cytosol, indicating that an early step in the import pathway was impeded. In a tobacco AS14 line expressing AS2.2, GFP was located in the cytosol, on the envelope, and in the stroma. The three patterns were observed in different cells, suggesting that the import capacity of individual cells was not the same. Our in vivo studies demonstrate that SPP is essential for chloroplast biogenesis and plant survival. SPP does not act independently in the stroma, but its activity influences earlier steps in the import pathway.
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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 | 0.001 | 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 | 0.001 | 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".