Maize Growth Promotion by Inoculation with an Engineered Ammonium-Excreting Strain of Nitrogen-Fixing Pseudomonasstutzeri
Abstract
:1. Introduction
2. Materials and Methods
2.1. Bacterial Strains and Media
2.2. In Vitro Screening for Plant Growth-Promoting Traits
2.3. Nitrogenase Activity Assays
2.4. Ammonium Excretion Determination
2.5. Pot Experiments and Inoculation Method
2.6. 15N Dilution Assay
2.7. Field Experiment for Crop Yield
2.8. Root Sampling and Bacterial Counts
2.9. Statistical Analysis
3. Results
3.1. Properties of the Wild-Type and Mutant Derivatives
3.2. Colonization of Wild-Type and Mutant Strains on the Maize Root Surface
3.3. Effects of Wild and Mutant Strains Inoculation on Maize Growth Promotion
3.4. Effects of Inoculation with P. stutzeri Strains on Maize N Concentrations
3.5. Crop Yield in Field Experiment
4. Discussion
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Strains | Mannitol-Treated Condition | Phosphate Solubilization Ability | IAA Production (mg/L) | ACC Deaminase Activity (μmol a-Ketobutyrate mg Protein−1 h−1) | Nitrogenase Activity (nmol of Ethylene mg Protein−1 h−1) | Extracellular Ammonium Concentration (μM) | ||
---|---|---|---|---|---|---|---|---|
Diameter of Halo (cm) | Diameter of Colony (cm) | PSI | ||||||
1568/pVA3 | 0 | 1.70 ± 0.01 a | 1.11 ± 0.02 a | 1.53 ± 0.04 a | 28.61 ± 1.11 b | 3.88 ± 0.10 a | 1792.3 ± 98.7 b | 20.3 ± 0.4 b |
200 mM | 1.74 ± 0.02 a | 1.13 ± 0.02 a | 1.54 ± 0.06 a | 32.67 ± 0.99 c | 3.74 ± 0.06 a | 1675.1 ± 112.9 ab | 18.2 ± 0.2 a | |
A1501 | 0 | 1.68 ± 0.04 a | 1.06 ± 0.03 a | 1.59 ± 0.12 a | 22.88 ± 1.17 a | 3.80 ± 0.09 a | 1618.2 ± 101.4 ab | n.d. |
200 mM | 1.72 ± 0.02 a | 1.10 ± 0.02 a | 1.57 ± 0.07 a | 29.26 ± 2.47 b | 3.85 ± 0.11 a | 1564.8 ± 90.6 a | n.d. | |
nifH-mutant | 0 | 1.69 ± 0.03 a | 1.12 ± 0.01 a | 1.51 ± 0.05 a | 24.79 ± 2.30 a | 3.76 ± 0.05 a | n.d. | n.d. |
200 mM | 1.73 ± 0.03 a | 1.12 ± 0.04 a | 1.55 ± 0.10 a | 28.14 ± 1.03 b | 3.81 ± 0.07 a | n.d. | n.d. |
Inoculant Treatments | SW (g) | RW (g) | SL (cm) | RL (cm) | Bacteria Root Colonization (CFU g−1) | |||||
---|---|---|---|---|---|---|---|---|---|---|
WS | WW | WS | WW | WS | WW | WS | WW | WS | WW | |
1568/pVA3 | 22.7 ± 0.9 b | 53.1 ± 2.8 c | 5.1 ± 0.2 c | 10.4 ± 0.9 c | 61.1 ± 4.2 b | 74.3 ± 2.8 b | 21.2 ± 3.8 a | 30.7 ± 3.5 a | 8.15 × 105 b | 1.54 × 107 b |
A1501 | 21.2 ± 0.8 b | 47.6 ± 3.9 b | 4.9 ± 0.3 bc | 9.8 ± 0.6 c | 59.7 ± 3.6 b | 67.6 ± 4.0 b | 20.9 ± 2.4 a | 29.8 ± 2.6 a | 9.09 × 105 b | 1.46 × 107 b |
nifH-mutant | 18.1 ± 1.0 a | 39.4 ± 2.7 a | 4.7 ± 0.1 b | 8.1 ± 0.3 b | 49.3 ± 3.0 a | 58.9 ± 1.8 a | 20.5 ± 3.9 a | 29.4 ± 3.3 a | 1.04 × 106 b | 1.38 × 107 b |
No inoculation | 17.3 ± 0.5 a | 37.4 ± 3.4 a | 4.3 ± 0.1 a | 6.8 ± 1.1 a | 48.2 ± 1.4 a | 57.4 ± 3.3 a | 20.1 ± 4.2 a | 29.1 ± 4.2 a | 7.02 × 102 a | 2.26 × 102 a |
Inoculant Treatments | Shoot N Concentrations (%) | Shoot Atom% 15N Excess (%) | % Ndfa | Total N Fixed (g/Plant) | ||||
---|---|---|---|---|---|---|---|---|
WS | WW | WS | WW | WS | WW | WS | WW | |
1568/pVA3 | 2.23 ± 0.06 b | 2.44 ± 0.07 b | 3.98 ± 0.12 b | 3.73 ± 0.16 b | 18.9 ± 1.2 b | 28.4 ± 0.9 b | 0.13 ± 0.01 b | 0.39 ± 0.04 b |
A1501 | 2.17 ± 0.03 ab | 2.38 ± 0.05 ab | 4.17 ± 0.20 b | 3.99 ± 0.22 b | 15.1 ± 0.7 a | 23.3 ± 1.0 a | 0.09 ± 0.01 a | 0.29 ± 0.02 a |
nifH-mutant | 2.18 ± 0.02 ab | 2.30 ± 0.06 a | 4.91 ± 0.13 a | 5.21 ± 0.10 a | - | - | - | - |
No inoculation | 2.12 ± 0.08 a | 2.31 ± 0.02 a | 5.10 ± 0.09 a | 5.34 ± 0.17 a | - | - | - | - |
Water Treatment | Inoculant Treatment | NH4+-N (mg/kg) | NO3−N (mg/kg) | Organic C (mg/kg) |
---|---|---|---|---|
WS | 1568/pVA3 | 10.7 ± 1.03 b | 5.42 ± 0.40 b | 10.43 ± 0.05 a |
A1501 | 2.47 ± 0.11 a | 3.91 ± 0.23 a | 11.10 ± 0.03 a | |
nifH-mutant | 2.61 ± 0.06 a | 4.04 ± 0.13 a | 10.56 ± 0.07 a | |
Non-inoculation | 2.50 ± 0.09 a | 3.90 ± 0.14 a | 11.02 ± 0.02 a | |
WW | 1568/pVA3 | 13.3 ± 1.26 b | 8.91 ± 0.51 b | 11.21 ± 0.10 a |
A1501 | 2.83 ± 0.07 a | 6.23 ± 0.62 a | 10.49 ± 0.06 a | |
nifH-mutant | 2.87 ± 0.10 a | 6.42 ± 0.98 a | 10.84 ± 1.02 a | |
Non-inoculation | 2.90 ± 0.08 a | 6.73 ± 0.80 a | 10.88 ± 0.09 a |
Inoculant Treatments | Nitrogen Application | SH (cm) | SDW (g/Plant) | NC (mg/g) | LL (cm) | WL (cm) | DS (cm) | RN (no./Plant) | GN (no./Plant) | TSW (g) | GY (t/ha) |
---|---|---|---|---|---|---|---|---|---|---|---|
0 N | 245.1 ± 2.0 ab | 232.1 ± 3.8 b | 18.1 ± 0.2 b | 97.9 ± 1.1 a | 9.7 ± 0.1 b | 2.76 ± 0.05 bc | 16.6 ± 0.2 ab | 37.1 ± 1.2 ab | 369.4 ± 10.0 b | 4.75 ± 0.04 b | |
1568/pVA3 | 75% N | 250.2 ± 1.4 c | 248.2 ± 1.3 c | 19.3 ± 0.2 c | 101.4 ± 1.5 b | 10.3 ± 0.2 b | 2.89 ± 0.07 c | 16.8 ± 0.3 b | 38.2 ± 1.0 b | 416.3 ± 7.5 cd | 10.65 ± 0.04 d |
100% N | 253.2 ± 1.8 d | 276.1 ± 4.2 f | 21.3 ± 0.3 d | 105.0 ± 1.0 c | 11.0 ± 0.1 c | 3.01 ± 0.08 d | 16.9 ± 0.1 b | 39.7 ± 1.3 bc | 443.7 ± 8.8 e | 12.15 ± 0.07 e | |
0 N | 242.8 ± 1.1 a | 230.0 ± 1.7 b | 17.4 ± 0.1 ab | 97.0 ± 1.5 a | 9.4 ± 0.1 a | 2.69 ± 0.04 b | 16.3 ± 0.2 a | 35.7 ± 0.9 a | 352.9 ± 9.1 a | 4.55 ± 0.03 a | |
A1501 | 75% N | 247.5 ± 2.9 bc | 251.6 ± 2.3 c | 18.9 ± 0.3 c | 99.5 ± 1.7 b | 9.9 ± 0.2 b | 2.80 ± 0.06 c | 16.4 ± 0.1 a | 37.8 ± 1.8 b | 402.5 ± 11.3 c | 10.15 ± 0.08 c |
100% N | 251.4 ± 1.0 cd | 278.3 ± 4.5 f | 19.9 ± 0.2 d | 104.6 ± 1.4 c | 10.9 ± 0.1 c | 2.93 ± 0.04 d | 16.8 ± 0.2 b | 38.9 ± 1.6 bc | 431.7 ± 9.2 e | 11.60 ± 0.09 e | |
0 N | 241.2 ± 1.2 a | 224.5 ± 1.6 a | 17.2 ± 0.1 a | 95.7 ± 1.2 a | 9.2 ± 0.1 a | 2.60 ± 0.07 a | 16.2 ± 0.1 a | 35.9 ± 1.5 a | 350.6 ± 12.0 a | 4.45 ± 0.05 a | |
nifH-mutant | 75% N | 249.9 ± 1.1 c | 259.1 ± 1.9 d | 18.8 ± 0.3 c | 98.3 ± 1.4 ab | 9.8 ± 0.1 b | 2.86 ± 0.02 c | 16.5 ± 0.1 a | 38.2 ± 2.0 bc | 402.5 ± 10.7 c | 9.95 ± 0.08 c |
100% N | 255.6 ± 2.7 d | 270.4 ± 3.2 e | 20.5 ± 0.2 d | 104.2 ± 1.0 c | 10.8 ± 0.2 c | 2.98 ± 0.04 d | 16.8 ± 0.3 b | 39.9 ± 1.1 bc | 434.7 ± 8.7 e | 12.00 ± 0.04 e | |
0 N | 242.6 ± 2.8 a | 228.5 ± 2.4 ab | 17.5 ± 0.1 a | 96.8 ± 1.0 a | 9.3 ± 0.1 a | 2.62 ± 0.03 a | 16.3 ± 0.1 a | 36.6 ± 1.3 a | 355.3 ± 11.2 a | 4.50 ± 0.05 a | |
No inoculation | 75% N | 249.1 ± 2.5 c | 258.1 ± 2.9 d | 19.0 ± 0.2 c | 99.3 ± 1.3 ab | 10.1 ± 0.1 b | 2.83 ± 0.05 c | 16.8 ± 0.3 b | 38.9 ± 1.8 bc | 410.1 ± 9.8 cd | 10.05 ± 0.06 c |
100% N | 254.4 ± 3.1 d | 279.8 ± 3.1 f | 20.7 ± 0.4 d | 103.1 ± 2.2 bc | 11.2 ± 0.1 c | 2.99 ± 0.02 d | 16.9 ± 0.2 b | 40.2 ± 2.1 c | 440.1 ± 10.5 e | 11.90 ± 0.08 e |
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Jiang, S.; Li, J.; Wang, Q.; Yin, C.; Zhan, Y.; Yan, Y.; Lin, M.; Ke, X. Maize Growth Promotion by Inoculation with an Engineered Ammonium-Excreting Strain of Nitrogen-Fixing Pseudomonasstutzeri. Microorganisms 2022, 10, 1986. https://doi.org/10.3390/microorganisms10101986
Jiang S, Li J, Wang Q, Yin C, Zhan Y, Yan Y, Lin M, Ke X. Maize Growth Promotion by Inoculation with an Engineered Ammonium-Excreting Strain of Nitrogen-Fixing Pseudomonasstutzeri. Microorganisms. 2022; 10(10):1986. https://doi.org/10.3390/microorganisms10101986
Chicago/Turabian StyleJiang, Shanshan, Jiang Li, Qingyu Wang, Changyan Yin, Yuhua Zhan, Yongliang Yan, Min Lin, and Xiubin Ke. 2022. "Maize Growth Promotion by Inoculation with an Engineered Ammonium-Excreting Strain of Nitrogen-Fixing Pseudomonasstutzeri" Microorganisms 10, no. 10: 1986. https://doi.org/10.3390/microorganisms10101986