AOX1a Expression in Arabidopsis thaliana Affects the State of Chloroplast Photoprotective Systems under Moderately High Light Conditions
Abstract
:1. Introduction
2. Results
2.1. Plants with Altered AOX1a Expression Exhibit Different Respiration Responses to MHL
2.2. Specific Leaf Weight and Chl Content
2.3. Effects of MHL on Photosynthetic Apparatus Functioning
2.4. Violaxanthin De-Epoxidation and the Effectiveness of the Violaxanthin Cycle Decreased in AS-12 Plants after 8 h of MHL
2.5. Violaxanthin De-Epoxidase Protein Content Follows the NPQ1 Expression
2.6. Changes in the Ascorbate Pool in Leaves and Chloroplasts of A. thaliana WT and Aox1a Lines after MHL
2.7. Gene Expression and Activity of Ascorbate Peroxidase in Leaves and Chloroplasts
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Plant Material and Growth Conditions
5.2. Specific Leaf Weight
5.3. O2 Exchange and CN-Resistant Respiration in Leaf Tissues
5.4. Gene Expression Analysis
5.5. SDS-PAGE and VDE Western Blot Analysis
5.6. Chl Extraction and Quantification
5.7. Carotenoid Analysis and Xanthophyll Cycle Activity Measurement
5.8. Chl Fluorescence Measurements
5.9. Measurement of Ascorbate and Dehydroascorbate Levels
5.10. Analysis of Ascorbate Peroxidase Activity
5.11. Chloroplast Isolation
5.12. Determination of Chl and Protein in Isolated Chloroplasts
5.13. Determination of Ascorbate and Dehydroascorbate Content in Isolated Chloroplasts
5.14. Measurement of Ascorbate Peroxidase Activity in Isolated Chloroplasts
5.15. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene Name | AGI No. | Gene Description ** | Forward Primer (5′~3′) Reverse Primer (5′~3′) | Product Length, bp |
---|---|---|---|---|
NPQ1 | At1g08550 | Non-photochemical quenching 1 | CGAGTGTGCTGTGTCGAGAA TGGCAGTCGAAGGCATCAAA | 168 |
APX1 | At1g07890 | Ascorbate peroxidase 1, cytosolic | TGGCCCTGACATTCCTTTCC ATGTGGGCCTCAGCGTAATC | 393 |
SAPX | At4g08390 | Stromal ascorbate peroxidase | GCGTCGGTGAATCGGAGTTT CCACCTCTTTGTGGCCATTC | 135 |
AT2G28390 * | At2g28390 | SAND family protein | AACTCTATGCAGCATTTGATCCACT TGATTGCATATCTTTATCGCCATC | 61 |
AT4G34270 * | At4g34270 | TIP41-like family protein | GTGAAAACTGTTGGAGAGAAGCAA TCAACTGGATACCCTTTCGCA | 61 |
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Garmash, E.V.; Dymova, O.V.; Silina, E.V.; Malyshev, R.V.; Belykh, E.S.; Shelyakin, M.A.; Velegzhaninov, I.O. AOX1a Expression in Arabidopsis thaliana Affects the State of Chloroplast Photoprotective Systems under Moderately High Light Conditions. Plants 2022, 11, 3030. https://doi.org/10.3390/plants11223030
Garmash EV, Dymova OV, Silina EV, Malyshev RV, Belykh ES, Shelyakin MA, Velegzhaninov IO. AOX1a Expression in Arabidopsis thaliana Affects the State of Chloroplast Photoprotective Systems under Moderately High Light Conditions. Plants. 2022; 11(22):3030. https://doi.org/10.3390/plants11223030
Chicago/Turabian StyleGarmash, Elena V., Olga V. Dymova, Ekaterina V. Silina, Ruslan V. Malyshev, Elena S. Belykh, Mikhail A. Shelyakin, and Ilya O. Velegzhaninov. 2022. "AOX1a Expression in Arabidopsis thaliana Affects the State of Chloroplast Photoprotective Systems under Moderately High Light Conditions" Plants 11, no. 22: 3030. https://doi.org/10.3390/plants11223030