Grape SnRK2.7 Positively Regulates Drought Tolerance in Transgenic Arabidopsis
Abstract
:1. Introduction
2. Results Analysis
2.1. Evolutionary Analysis of VvSNRK2 Genes
2.2. Analysis of Physicochemical Properties of VvSnRK2.7
2.3. Subcellular Localization of VvSNRK2.7
2.4. Yeast Library Screening and PCR Assay
2.5. Validation of VvSnRK2.7 Interactions with bZIP Protein
2.6. pCAMBIA1300-VvSnRK2.7 Overexpression Vector Construction and Characterization of Transgenic Arabidopsis
2.7. Morphological Observation and Expression Analysis of Transgenic Arabidopsis after Drought Stress
2.8. VvSnRK2.7 Increased Antioxidant Enzyme Activities and Proline Content under Drought Stress
2.9. Overexpression of the VvSnRK2.7 Alters Arabidopsis Leaf MDA Content and Relative Conductance
2.10. Overexpression of VvSnRK2.7 Increases Leaf Sugar Content in Arabidopsis
3. Discussion
4. Materials and Methods
4.1. Test Materials
4.2. Obtaining and Cloning of Grape VvSnRK2.7
4.3. Overexpression and Subcellular Localization of Recombinant Plasmid Transformation
4.4. Functional Analysis of Drought Resistance in Transgenic Arabidopsis
4.5. Measurement of Relevant Indicators after Stress
4.6. Screening of VvSnRK2.7-Interacting Proteins with a Mating Assay
4.7. Validation of the VvSnRK2.7 with VvbZIP
4.8. qRT-PCR Analysis
4.9. Data Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Gene | Amino Acid Number | Molecular Weight | Isoelectric Point | Full Length/bp | Instability Index |
---|---|---|---|---|---|
PNVvSnRK2.7 | 355 | 40.61 | 6.05 | 5680 | 45.07 |
RGVvSnRK2.1 | 355 | 40.62 | 6.1 | 5683 | 47.12 |
ChVvSnRk2.1 | 355 | 40.68 | 6.05 | 5690 | 47.12 |
Gene | Primer Sequence (5′-3′) | Purpose |
---|---|---|
VvSnRK2.7-EGFP | 5′-gagctcggtacccggggatccATGGAGAAGTATGAAATGGTGAAGG | Transient expression |
3′-ggtgtcgactctagaggatccACTAACGTGAAATTCTCCGCTTG | Transient expression | |
pCAMBIA1300-VvSnRK2.7 | 5′-gagctcggtacccggggatccATGGAGAAGTATGAAATGGTGAAGG | Overexpression |
3′-ggtgtcgactctagaggatccTTAACTAACGTGAAATTCTCCGCT | Overexpression | |
pGBKT7-VvSnRK2.7 | 5′-aggccgaattcccggggatccttATGGAGAAGTATGAAATGGTGAAGG | BD vector primers |
3′-ccgctgcaggtcgacggatccTTAACTAACGTGAAATTCTCCGCT | BD vector primers | |
pGADT7-VvbZIP | 5′-gtgggcatcgatacgggatccatATGTTGTCATCAACAGGTGGCG | AD vector primers |
3′-cagctcgagctcgatggatccTCAAAATGGGGCTGTTGAAGTT | AD vector primers | |
qRT-AtActin | 5′-CTTGCACCAAGCAGCATGAA | qRT-PCR primers |
3′-CCGATCCAGACACTGTACTTCCTT | qRT-PCR primers | |
qRT-VvSnRK2.7 | 5′-AGAAGAGGGTAAAGGCGGTGAGG | qRT-PCR primers |
3′-R-CGCTTGCATGGACTTCCCTAACC | qRT-PCR primers |
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Lan, G.; Ma, W.; Nai, G.; Liang, G.; Lu, S.; Ma, Z.; Mao, J.; Chen, B. Grape SnRK2.7 Positively Regulates Drought Tolerance in Transgenic Arabidopsis. Int. J. Mol. Sci. 2024, 25, 4473. https://doi.org/10.3390/ijms25084473
Lan G, Ma W, Nai G, Liang G, Lu S, Ma Z, Mao J, Chen B. Grape SnRK2.7 Positively Regulates Drought Tolerance in Transgenic Arabidopsis. International Journal of Molecular Sciences. 2024; 25(8):4473. https://doi.org/10.3390/ijms25084473
Chicago/Turabian StyleLan, Guanquecailang, Weifeng Ma, Guojie Nai, Guoping Liang, Shixiong Lu, Zonghuan Ma, Juan Mao, and Baihong Chen. 2024. "Grape SnRK2.7 Positively Regulates Drought Tolerance in Transgenic Arabidopsis" International Journal of Molecular Sciences 25, no. 8: 4473. https://doi.org/10.3390/ijms25084473