Morphological and Physiological Responses of In Vitro-Grown Cucurbita sp. Landraces Seedlings under Osmotic Stress by Mannitol and PEG
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Growth Condition, Experimental Design, and Treatments
2.2. Hypocotyl Length, Shoot and Root Fresh and Dry Weight
2.3. Photosynthetic Pigments
2.4. Total Proline Content
2.5. Malondialdehyde (MDA) Content
2.6. Total Soluble Protein Content
2.7. Total Phenol Content (TPC)
2.8. Total Flavonoids Content (TFC)
2.9. H2O2 Content
2.10. Enzymatic Antioxidants Activity
2.10.1. Ascorbate Peroxidase Activity (APX)
2.10.2. Guaiacol Peroxidase Activity (GPX)
2.10.3. Superoxide Dismutase Activity (SOD)
2.10.4. Glutathione Reductase (GR) Activity
2.11. Non-Enzymatic Antioxidants Activity
2.12. Statistical Analysis
3. Results
3.1. Hypocotyl Length
3.2. Shoot and Root FW and DW
3.3. Photosynthesis Pigments
3.4. Total Proline Content
3.5. Total Soluble Protein Content
3.6. Total Phenol and Flavonoid Content
3.7. Enzymatic Antioxidant Activity
3.8. Malondialdehyde Content
3.9. Hydrogen Peroxide Content
3.10. Nonenzymatic Antioxidants
3.11. Multivariate Analysis of Cucurbits Landraces under Normal and Mannitol and PEG Treatments
4. Discussion
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Species | Osmotic Agent | Concentration (M) | Plant Characters | ||||
---|---|---|---|---|---|---|---|
Hypocotyl Length (mm) | Shoot FW (g Seedling−1) | Shoot DW (%) | Root FW (g Seedling−1) | Root DW (%) | |||
BG | Control | 0 | 24.38 ± 0.33 gh | 2.71 ± 0.20 a | 7.78 ± 0.40 k | 0.71 ± 0.11 a | 7.85 ± 0.30 g–i |
Mannitol | 0.1 | 18.05 ± 1.29 h–j | 2.02 ± 0.29 b | 12.57 ± 1.05 f | 0.20 ± 0.03 ij | 9.09 ± 0.50 ef | |
0.2 | 13.47 ± 1.32 jk | 1.45 ± 0.24 cd | 18.13 ± 0.47 bc | 0.15 ± 0.03 j | 14.18 ± 0.33 a | ||
0.4 | 7.20 ± 1.30 k | 1.12 ± 0.21 e–g | 20.66 ± 0.77 a | 0.13 ± 0.02 j | 13.52 ± 0.96 ab | ||
PEG | 0.009 | 20.90 ± 0.88 hi | 2.50 ± 0.10 a | 9.23 ± 0.13 i–k | 0.52 ± 0.07 cd | 9.912 ± 0.45 de | |
0.012 | 17.69 ± 0.68 h–j | 1.75 ± 0.11 bc | 14.25 ± 1.17 de | 0.18 ± 0.04 ij | 11.31 ± 0.40 c | ||
0.015 | 15.06 ± 0.47 ij | 1.26 ± 0.12 d–f | 16.69 ± 0.33 c | 0.13 ± 0.02 j | 12.83 ± 0.42 b | ||
TA | Control | 0 | 70.72 ± 2.30 b | 1.65 ± 0.09 c | 9.04 ± 0.80 jk | 0.70 ± 0.06 a | 5.68 ±0.19 l |
Mannitol | 0.1 | 43.73 ± 4.46 cd | 1.07 ± 0.07 e–g | 11.96 ± 0.32 fg | 0.30 ± 0.09 f–i | 7.04 ± 0.64 i–k | |
0.2 | 36.10 ± 5.83 ef | 1.00 ± 0.06 f–h | 12.71 ± 0.73 f | 0.25 ± 0.07 g–j | 9.17 ± 0.64 ef | ||
0.4 | 13.05 ± 3.71 jk | 0.69 ± 0.10 i | 17.50 ± 1.08 bc | 0.12 ± 0.03 j | 10.79 ± 0.59 cd | ||
PEG | 0.009 | 69.83 ± 1.69 b | 1.45 ± 0.08 cd | 10.66 ± 0.34 g–i | 0.66 ± 0.11 ab | 6.25 ± 0.44 j–l | |
0.012 | 33.46 ± 6.56 ef | 1.11 ± 0.10 e–g | 12.92 ± 0.98 ef | 0.37 ± 0.05 e–g | 8.19 ± 0.63 f–i | ||
0.015 | 12.06 ± 1.70 jk | 0.63 ± 0.09 i | 18.82 ± 0.41 b | 0.21 ± 0.04 ij | 11.03 ± 0.96 c | ||
RA | Control | 0 | 79.63 ± 8.64 a | 1.66 ± 0.13 c | 7.90 ± 0.42 k | 0.63 ± 0.02 a–c | 6.15 ± 0.61 kl |
Mannitol | 0.1 | 39.52 ± 1.07 de | 1.22 ± 0.14 d–f | 9.79 ± 0.71 h–j | 0.54 ± 0.00 b–d | 7.31 ± 0.43 h–j | |
0.2 | 36.64 ± 2.32 d–f | 1.05 ± 0.13 e–g | 10.85 ± 1.03 gh | 0.47 ± 0.02 de | 8.12 ± 0.23 f–i | ||
0.4 | 29.43 ± 0.65 fg | 0.87 ± 0.06 g–i | 12.57 ± 0.16 f | 0.34 ± 0.03 f–h | 9.61 ± 0.39 e | ||
PEG | 0.009 | 47.99 ± 3.47 c | 1.34 ± 0.07 de | 9.81 ± 0.43 h–j | 0.55 ± 0.04 b–d | 5.96 ± 0.38 kl | |
0.012 | 38.30 ± 1.93 de | 1.08 ± 0.12 e–g | 12.52 ±1.00 f | 0.39 ± 0.10 ef | 8.33 ± 0.19 f–h | ||
0.015 | 24.91 ± 2.22 gh | 0.73 ± 0.04 hi | 15.20 ± 0.56 d | 0.24 ± 0.04 h–j | 8.95 ± 0.33 e–g | ||
S.O.V. | |||||||
Species Osmotic agent Species × Osmotic agent Error | 4207.017 ** | 3.642 ** | 48.868 ** | 0.141 ** | 73.052 ** | ||
2035.468 ** | 1.614 ** | 98.948 ** | 0.312 ** | 31.913 ** | |||
368.631 * | 0.096 ** | 10.950 * | 0.025 ** | 2.185 ** | |||
16.472 | 0.028 | 0.753 | 0.005 | 0.406 | |||
C.V. (%) | 12.31 | 12.26 | 6.71 | 19.08 | 7.00 |
Species | Osmotic Agent | Concentration (M) | Character | |||
---|---|---|---|---|---|---|
Chl a (mg kg−1 FW) | Chl b (mg kg−1 FW) | Chl a + b (mg kg−1 FW) | CARs (mg kg−1 FW) | |||
BG | Control | 0 | 34.43 ± 0.44 a | 21.86 ± 2.38 a | 56.29 ± 2.80 a | 8.68 ± 0.33 a |
Mannitol | 0.1 | 31.53 ± 3.27 b | 17.98 ± 0.59 b | 49.51 ± 3.55 b | 5.88 ± 1.04 c–e | |
0.2 | 23.87 ± 0.87 c | 15.48 ± 0.61 c | 39.35 ± 1.26 c | 4.83 ± 0.33 ef | ||
0.4 | 20.17 ± 0.27 d | 7.94 ± 0.02 g | 28.12 ± 0.26 e | 3.30 ± 0.32 hi | ||
PEG | 0.009 | 34.23 ± 0.53 a | 15.68 ± 0.99 c | 49.91 ± 0.56 b | 6.96 ± 0.69 bc | |
0.012 | 23.47 ± 0.93 c | 12.52 ± 0.99 d | 36.00 ± 0.91 d | 3.29 ± 0.15 hi | ||
0.015 | 19.79 ± 0.50 d | 9.71± 0.36 ef | 29.51 ± 0.76 e | 2.58 ± 0.29 ij | ||
TA | Control | 0 | 13.44 ± 0.91 f | 8.39 ± 0.42 fg | 21.84 ± 0.54 fg | 9.25± 0.57 a |
Mannitol | 0.1 | 10.99 ± 0.49 g | 8.09 ± 0.44 fg | 19.09 ± 0.93 g | 7.04 ± 0.64 bc | |
0.2 | 7.84 ± 0.76 h–j | 4.83 ± 0.56 hi | 12.68 ± 0.47 hi | 4.61 ± 0.99 e–g | ||
0.4 | 4.10 ± 0.68 kl | 2.67 ± 0.57 jk | 6.77 ± 1.10 kl | 1.94 ± 0.14 j | ||
PEG | 0.009 | 8.78 ± 0.66 g–i | 6.03 ± 0.71 h | 14.81 ± 1.27 h | 6.82 ± 0.26 b–d | |
0.012 | 6.06 ± 0.46 jk | 3.02 ± 0.33 jk | 9.08 ± 0.79 jk | 4.96 ± 0.57 ef | ||
0.015 | 3.22 ± 0.17 l | 1.72 ± 0.23 k | 4.94 ± 0.41 l | 2.45 ± 0.16 ij | ||
RA | Control | 0 | 30.44 ± 2.41 b | 17.77 ± 0.45 b | 48.21 ± 2.57 b | 7.50 ± 1.41 b |
Mannitol | 0.1 | 25.28 ±0.51 c | 14.79 ± 0.55 c | 40.08 ± 0.53 c | 5.61 ± 0.28 de | |
0.2 | 16.17 ± 0.71 e | 8.10 ± 0.25 fg | 24.27 ± 0.95 f | 3.45 ± 0.05 g–i | ||
0.4 | 6.90 ± 0.35 ij | 3.83 ± 0.51 ij | 10.74 ± 0.84 ij | 2.49 ± 0.60 ij | ||
PEG | 0.009 | 20.63 ± 0.56 d | 10.38 ± 0.88 e | 31.01 ± 1.39 e | 6.31 ± 0.53 b–d | |
0.012 | 13.96 ± 0.72 ef | 8.45 ± 0.80 fg | 22.41 ± 1.27 f | 4.21 ± 0.35 f–h | ||
0.015 | 9.69 ± 1.85 gh | 2.38 ± 0.06 jk | 12.08 ± 1.80 h–j | 1.98 ± 0.14 j | ||
S.O.V. | ||||||
Species | 1896.903 ** | 473.298 ** | 4262.121 ** | 3.489 ** | ||
Osmotic agent | 328.683 ** | 163.292 ** | 946.994 ** | 45.801 ** | ||
Species × Osmotic agent | 24.221 ** | 9.261 ** | 51.632 ** | 1.077 * | ||
Error | 1.919 | 0.886 | 3.150 | 0.490 | ||
C.V. (%) | 7.97 | 9.80 | 6.58 | 14.10 |
Species | Osmotic Agent | Concentration (M) | Character | |||
---|---|---|---|---|---|---|
Proline (µm g−1 FW) | Total Protein (mg g−1 FW) | TPC (mg g−1 DW) | TFC (mg g−1 DW) | |||
BG | Control | 0 | 6.75 ± 0.13 gh | 1.60 ± 0.12 b–e | 33.67 ± 1.25 gh | 5 ± 0.82 de |
Mannitol | 0.1 | 8.21 ± 1.44 fg | 1.73 ± 0.22 a–d | 56 ± 2.45 a | 11.67 ± 1.25 a | |
0.2 | 12.26 ± 0.92 c–e | 1.97 ± 0.21 a | 46 ± 1.41 c | 5.333 ± 0.47 d | ||
0.4 | 16.17 ± 1.68 b | 1.56 ± 0.39 c–e | 29.67 ± 1.25 ij | 2.167 ± 0.45 g–i | ||
PEG | 0.009 | 10.30 ± 0.71 ef | 1.80 ± 0.20 a–c | 51 ± 2.16 b | 9.667 ± 0.47 b | |
0.012 | 14.76 ± 1.91 b–d | 1.98 ± 0.26 a | 41.67 ± 1.25 de | 2.333 ± 0.47 gh | ||
0.015 | 4.23 ± 0.69 h | 0.81 ± 0.05 h–j | 25.67 ± 2.49 kl | 1.433 ± 0.17 h–j | ||
TA | Control | 0 | 6.83 ± 1.83 gh | 0.81 ± 0.01 h–j | 27 ± 2.16 jk | 3.733 ± 0.38 f |
Mannitol | 0.1 | 12.92 ± 0.54 b–e | 0.99 ± 0.01 g–i | 41 ± 1.41 de | 6.667 ± 0.47 c | |
0.2 | 15.45 ± 0.39 bc | 1.30 ± 0.05 e–g | 33.67 ± 1.70 gh | 4.067 ± 0.25 ef | ||
0.4 | 24.11 ± 4.71 a | 1.11 ± 0.09–h | 20 ± 0.82 mn | 1.533 ± 0.21 h–j | ||
PEG | 0.009 | 8.00 ± 0.81 fg | 1.29 ± 0.17 e–g | 38.33 ± 0.94 ef | 5.400 ± 0.43 d | |
0.012 | 13.10 ± 0.89 b–e | 1.33 ± 0.15 e–g | 22.67 ± 1.70 lm | 2.633 ± 0.45 g | ||
0.015 | 4.52 ± 1.89 h | 0.82 ± 0.05 h–j | 17.67 ± 1.25 n | 1.167 ± 0.13 ij | ||
RA | Control | 0 | 5.96 ± 0.71 gh | 0.61 ± 0.15 j | 36 ± 1.63 fg | 4.267 ± 0.38 ef |
Mannitol | 0.1 | 11.57 ± 0.41 de | 1.34 ± 0.05 e–g | 49.67 ± 0.47 b | 5.7 ± 0.36 d | |
0.2 | 12.48 ± 0.33 c–e | 1.92 ± 0.12 ab | 43 ± c2.45 d | 3.8 ± 0.16 f | ||
0.4 | 14.46 ± 0.68 b–d | 1.07 ± 0.07 g–i | 32.33 ± 0.47 g–i | 2.1 ± 0.22 g–i | ||
PEG | 0.009 | 10.05 ± 0.45 ef | 1.09 ± 0.15 gh | 45.67 ± 2.06 c | 4.3 ± 0.16 ef | |
0.012 | 11.87 ± 0.56 de | 1.45 ± 0.16 d–f | 32 ± 1.63 hi | 1.767 ± 0.21 g–j | ||
0.015 | 3.81 ± 0.65 h | 0.72 ± 0.06 ij | 23 ± 2.16 lm | 1.033 ± 0.13 j | ||
S.O.V. | ||||||
Species | 26.633 ** | 1.815 ** | 799.302 ** | 26.634 ** | ||
Osmotic agent | 198.032 ** | 0.865 ** | 850.106 ** | 56.207 ** | ||
Species × Osmotic agent | 15.362 ** | 0.182 ** | 21.431 ** | 5.423 ** | ||
Error | 3.113 | 0.036 | 4.270 | 0.316 | ||
C.V. (%) | 16.26 | 14.60 | 5.82 | 13.76 |
Mean Square | ||||||
---|---|---|---|---|---|---|
df | CAT Activity | GR Activity | APX Activity | GPX Activity | SOD Activity | |
Species | 2 | 8.137 ** | 0.595 ** | 0.756 ** | 0.027 ns | 1449.242 ** |
Osmotic agent | 6 | 4.809 ** | 0.252 ** | 0.237 ** | 2.464 ** | 850.888 ** |
Species × Osmotic agent | 12 | 0.092 ** | 0.011 ** | 0.011 ** | 0.043 * | 111.711 ** |
Error | 42 | 0.016 | 0.001 | 0.002 | 0.022 | 7.190 |
C.V. (%) | 5.10 | 8.42 | 9.03 | 9.48 | 5.94 |
Mean Square | |||||||||
---|---|---|---|---|---|---|---|---|---|
df | AsA | DHA | ASA/DHA | GSH | GSSG | GSH/GSSG | H2O2 | MDA | |
Species | 2 | 120,832.68 ** | 56,691.48 | 36.608 ** | 91879 ** | 24,595.85 ** | 16.07 ** | 19.583 ** | 12.708 ** |
Osmotic agent | 6 | 9104.22 ** | 15,851.95 | 9.536 ** | 10,168.21 ** | 5246.95 ** | 2.711 ** | 5.294 ** | 3.335 ** |
Species × Osmotic agent | 12 | 756.35 ** | 896.86 | 2.373 ** | 682.037 ** | 240.63 ** | 0.095 ** | 0.314 ** | 0.323 ** |
Error | 42 | 13.952 | 14.63 | 0.019 | 9.095 | 11.06 | 0.004 | 0.013 | 0.026 |
C.V. (%) | 2 | 2.71 | 7.39 | 0.99 | 2.05 | 2.90 | 4.64 | 9.12 |
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Tajaragh, R.P.; Rasouli, F.; Giglou, M.T.; Zahedi, S.M.; Hassanpouraghdam, M.B.; Aazami, M.A.; Adámková, A.; Mlček, J. Morphological and Physiological Responses of In Vitro-Grown Cucurbita sp. Landraces Seedlings under Osmotic Stress by Mannitol and PEG. Horticulturae 2022, 8, 1117. https://doi.org/10.3390/horticulturae8121117
Tajaragh RP, Rasouli F, Giglou MT, Zahedi SM, Hassanpouraghdam MB, Aazami MA, Adámková A, Mlček J. Morphological and Physiological Responses of In Vitro-Grown Cucurbita sp. Landraces Seedlings under Osmotic Stress by Mannitol and PEG. Horticulturae. 2022; 8(12):1117. https://doi.org/10.3390/horticulturae8121117
Chicago/Turabian StyleTajaragh, Rana Panahi, Farzad Rasouli, Mousa Torabi Giglou, Seyed Morteza Zahedi, Mohammad Bagher Hassanpouraghdam, Mohammad Ali Aazami, Anna Adámková, and Jiří Mlček. 2022. "Morphological and Physiological Responses of In Vitro-Grown Cucurbita sp. Landraces Seedlings under Osmotic Stress by Mannitol and PEG" Horticulturae 8, no. 12: 1117. https://doi.org/10.3390/horticulturae8121117