Influence of Water and Fertilizer Reduction on Respiratory Metabolism in Sugar Beet Taproot (Beta vulgaris L.)
Abstract
:1. Introduction
2. Results
2.1. Sugar Beet Taproot Growth
2.2. Respiration Rate of Sugar Beet Taproot
2.3. Energy Metabolism of Sugar Beet Taproot
2.3.1. Contents of ATP, ADP, and AMP
2.3.2. Energy Charge
2.3.3. ATPase
2.4. Key Enzymes in Respiratory Metabolism of Sugar Beet Taproot
2.4.1. Hexokinase
2.4.2. Isocitrate Dehydrogenase
2.4.3. Glucose-6-Phosphate Dehydrogenase
2.5. Correlation Analysis between Taproot Fresh Weight and Respiratory Metabolism Indicators
3. Discussion
4. Materials and Methods
4.1. Experimental Site Description
4.2. Experimental Design
4.3. Collection of Plant Samples
4.4. Respiration Rate
4.5. Respiratory Metabolic Enzyme Activity and Energy Measurement
- (1)
- Standard Addition: Set up wells for the standards and samples. Add 50 μL of standards with different concentrations to each standard well.
- (2)
- Sample Addition: Set up blank wells (i.e., wells without the samples and enzyme reagents, but with all other steps remaining the same) and wells for the test samples. Add 40 μL of sample diluent to each test sample well on the coated microplate, followed by 10 μL of the test sample (resulting in a final dilution of 5 times). Gently mix.
- (3)
- Enzyme Addition: Add 100 μL of enzyme conjugate to each well except the blank wells.
- (4)
- Incubation: Cover the plate with a sealing film and incubate at 37 °C for 60 min.
- (5)
- Washing: Carefully remove the sealing film and discard the liquid. Fill each well with washing solution, let it stand for 30 s, and then discard the washing solution. Repeat this step five times and pat dry.
- (6)
- Color Development: Add 50 μL of chromogen solution A to each well, followed by 50 μL of chromogen solution B. Gently shake to mix, and incubate at 37 °C in the dark for 15 min.
- (7)
- Termination: Add 50 μL of stop solution to each well to terminate the reaction (this will cause the color to change from blue to yellow).
- (8)
- Measurement: Set the blank well to zero, and measure the absorbance (OD value) of each well at 450 nm. This should be completed within 15 min after adding the stop solution.
- (9)
- Calculation: Using the concentration and OD values of the standards, create a standard curve with a linear regression equation. Insert the OD values of the samples into this equation to calculate their concentrations, then multiply by the dilution factor to obtain the actual concentration of the samples.
4.6. Energy Charge Calculation Formula
4.7. Total RNA Extraction and cDNA Synthesis from Sugar Beets
4.8. Real-Time Quantitative PCR
4.9. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Year | Total N (g·kg−1) | Total P (g·kg−1) | Total K (g·kg−1) | Available N (mg·kg−1) | Available P (mg·kg−1) | Available K (mg·kg−1) | pH Value | Organic Matter (g·kg−1) |
---|---|---|---|---|---|---|---|---|
2022 | 2.07 | 0.97 | 22.11 | 134.91 | 10.31 | 197.5 | 7.98 | 32.02 |
2023 | 1.71 | 0.46 | 16.31 | 111.07 | 9.23 | 153.01 | 7.71 | 18.21 |
Treatment | Amount of Fertilizer Applied (kg/ha) | Total Irrigation Volume (m3/ha) | ||
---|---|---|---|---|
Pure N | Pure P | Pure K | ||
F3W3(CK) | 120 | 150 | 150 | 1350 |
F3W2 | 1147.5 | |||
F3W1 | 945 | |||
F2W3 | 108 | 135 | 135 | 1350 |
F2W2 | 1147.5 | |||
F2W1 | 945 | |||
F1W3 | 96 | 120 | 120 | 1350 |
F1W2 | 1147.5 | |||
F1W1 | 945 |
Primer | R 5′ to 3′ | F 5′ to 3′ |
---|---|---|
ACT | TGCTTGACTCTGGTGATGGT | AGCAAGATCCAAACGGAGAATG |
HK | GCGACGTTATGGTTGTAT | ACTCTTCCAACTCCCTCAACAC |
IDH | GCAACCATCACACCAGATGAA | ACTCTTCCAACTCCCTCAACAC |
G6PDH | ATGGATCTCCCTACCAGTCG | CTCATCAAAATAGCCACCTCT |
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Chang, Y.; Li, G.; Jian, C.; Zhang, B.; Sun, Y.; Li, N.; Zhang, S. Influence of Water and Fertilizer Reduction on Respiratory Metabolism in Sugar Beet Taproot (Beta vulgaris L.). Plants 2024, 13, 2282. https://doi.org/10.3390/plants13162282
Chang Y, Li G, Jian C, Zhang B, Sun Y, Li N, Zhang S. Influence of Water and Fertilizer Reduction on Respiratory Metabolism in Sugar Beet Taproot (Beta vulgaris L.). Plants. 2024; 13(16):2282. https://doi.org/10.3390/plants13162282
Chicago/Turabian StyleChang, Yuxin, Guolong Li, Caiyuan Jian, Bowen Zhang, Yaqing Sun, Ningning Li, and Shaoying Zhang. 2024. "Influence of Water and Fertilizer Reduction on Respiratory Metabolism in Sugar Beet Taproot (Beta vulgaris L.)" Plants 13, no. 16: 2282. https://doi.org/10.3390/plants13162282