Chinese Bulletin of Botany ›› 2020, Vol. 55 ›› Issue (1): 49-61.DOI: 10.11983/CBB19047
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Dongdong Cao1,3,Shanyu Chen1,Yebo Qin2,Huaping Wu3,Guanhai Ruan1,Yutao Huang1,*(
)
HomeReceived:2019-03-14
Accepted:2019-07-26
Online:2020-01-01
Published:2019-12-20
Contact:
Yutao Huang
Dongdong Cao,Shanyu Chen,Yebo Qin,Huaping Wu,Guanhai Ruan,Yutao Huang. Regulatory Mechanism of Salicylic Acid on Seed Germination Under Salt Stress in Kale[J]. Chinese Bulletin of Botany, 2020, 55(1): 49-61.
| Gene name | No. of accession | Primer sequence (5'-3') |
|---|---|---|
| 18srRNA | NC_016118.1 | F: GGAAGGACTTGTACGGTAACATTG R: TGGACCTGCCTCATCATACTCA |
| BoPAL1 | XM_013781025.1 | F: TCGATCTTCCACAAGATTGGT R: TCCACTTCGTCAGGAAGCA |
| BoPAL2 | XM_013749252.1 | F: AATCAGCTGAGCAACATAACCA R: GACGTTTTGCGAGACGAGA |
| BoCAT1 | XM_013742034.1 | F: ATCCTCGTGGTTTTGCTGTC R: TGCCAACAAGATCAAAGTTCC |
| BoCAT2 | XM_013774413.1 | F: ATGGAAGGCTCAGGTGTCAA R: CGTAGTGAGCTTTTCCGGATT |
| BoSOD1 | XM_013783393.1 | F: ACATCATTGTTGGAGATGATGG R: GAGGGATCTGGCAGTCAGTG |
| BoSOD3 | XM_013750980.1 | F: TGGTGATCCTGATGACCTTG R: CCTGCGTTTCCTGTTGATTT |
| BoPOD2 | XM_013729485.1 | F: GGTGGTTTCTTGCGCTGA R: CCATGGTCCGTTGATCACTA |
| BoPOD3 | XM_013728218.1 | F: CGTCAGGAAACACGGAGAA R: CCTCAATGAAGCCAAATCCT |
Table 1 Primers used for qRT-PCR
| Gene name | No. of accession | Primer sequence (5'-3') |
|---|---|---|
| 18srRNA | NC_016118.1 | F: GGAAGGACTTGTACGGTAACATTG R: TGGACCTGCCTCATCATACTCA |
| BoPAL1 | XM_013781025.1 | F: TCGATCTTCCACAAGATTGGT R: TCCACTTCGTCAGGAAGCA |
| BoPAL2 | XM_013749252.1 | F: AATCAGCTGAGCAACATAACCA R: GACGTTTTGCGAGACGAGA |
| BoCAT1 | XM_013742034.1 | F: ATCCTCGTGGTTTTGCTGTC R: TGCCAACAAGATCAAAGTTCC |
| BoCAT2 | XM_013774413.1 | F: ATGGAAGGCTCAGGTGTCAA R: CGTAGTGAGCTTTTCCGGATT |
| BoSOD1 | XM_013783393.1 | F: ACATCATTGTTGGAGATGATGG R: GAGGGATCTGGCAGTCAGTG |
| BoSOD3 | XM_013750980.1 | F: TGGTGATCCTGATGACCTTG R: CCTGCGTTTCCTGTTGATTT |
| BoPOD2 | XM_013729485.1 | F: GGTGGTTTCTTGCGCTGA R: CCATGGTCCGTTGATCACTA |
| BoPOD3 | XM_013728218.1 | F: CGTCAGGAAACACGGAGAA R: CCTCAATGAAGCCAAATCCT |
Figure 1 Effects of salt stress on seed germination of kale (A) Time course of seed germination under salt stress; (B) Seed germination after 96 hours of NaCl treatments with different concentration
| Treatments | Water uptake rate (%) | |||||
|---|---|---|---|---|---|---|
| 0-4 h | 4-8 h | 8-16 h | 16-24 h | 24-36 h | 36-48 h | |
| Control-1 | 47.18±3.52 a | 42.56±2.19 a | 19.14±0.81 a | 18.33±2.04 a | 18.21±1.33 a | 27.26±2.67 a |
| Control-2 | 37.11±4.31 b | 31.67±3.15 c | 14.62±1.09 b | 12.62±0.09 c | 10.21±1.02 b | 17.64±0.08 b |
| SA | 42.27±2.89 a | 37.75±3.09 b | 14.31±1.59 b | 14.24±0.15 b | 16.77±1.44 a | 25.81±1.77 a |
| AIP | 32.12±2.71 c | 30.41±4.93 c | 13.41±1.07 c | 12.78±0.21 c | 8.21±0.06 c | 7.55±1.23 c |
Table 2 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on water uptake rate during seed germination of kale under salt stress
| Treatments | Water uptake rate (%) | |||||
|---|---|---|---|---|---|---|
| 0-4 h | 4-8 h | 8-16 h | 16-24 h | 24-36 h | 36-48 h | |
| Control-1 | 47.18±3.52 a | 42.56±2.19 a | 19.14±0.81 a | 18.33±2.04 a | 18.21±1.33 a | 27.26±2.67 a |
| Control-2 | 37.11±4.31 b | 31.67±3.15 c | 14.62±1.09 b | 12.62±0.09 c | 10.21±1.02 b | 17.64±0.08 b |
| SA | 42.27±2.89 a | 37.75±3.09 b | 14.31±1.59 b | 14.24±0.15 b | 16.77±1.44 a | 25.81±1.77 a |
| AIP | 32.12±2.71 c | 30.41±4.93 c | 13.41±1.07 c | 12.78±0.21 c | 8.21±0.06 c | 7.55±1.23 c |
| Treatments | Germination energy (%) | Germination percentage (%) | Germination index | Vigor index | Seedling length (cm) | Seedling dry weight (g) |
|---|---|---|---|---|---|---|
| Control-1 | 85.333±3.270 a | 85.333±3.272 a | 15.742±0.742 a | 1.610±0.892 a | 8.241±0.711 a | 0.083±0.010 a |
| Control-2 | 53.667±3.208 c | 56.333±4.044 c | 8.231±0.810 c | 0.908±0.071 c | 7.822±0.635 b | 0.076±0.008 b |
| SA | 62.333±4.615 b | 68.667±4.730 b | 10.558±0.556 b | 1.167±0.064 b | 7.837±0.706 b | 0.079±0.009 b |
| AIP | 41.000±3.331 d | 46.667±2.516 d | 7.267±0.597 d | 0.762±0.079 d | 7.156±0.523 c | 0.064±0.004 c |
Table 3 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on seed germination of kale under salt stress
| Treatments | Germination energy (%) | Germination percentage (%) | Germination index | Vigor index | Seedling length (cm) | Seedling dry weight (g) |
|---|---|---|---|---|---|---|
| Control-1 | 85.333±3.270 a | 85.333±3.272 a | 15.742±0.742 a | 1.610±0.892 a | 8.241±0.711 a | 0.083±0.010 a |
| Control-2 | 53.667±3.208 c | 56.333±4.044 c | 8.231±0.810 c | 0.908±0.071 c | 7.822±0.635 b | 0.076±0.008 b |
| SA | 62.333±4.615 b | 68.667±4.730 b | 10.558±0.556 b | 1.167±0.064 b | 7.837±0.706 b | 0.079±0.009 b |
| AIP | 41.000±3.331 d | 46.667±2.516 d | 7.267±0.597 d | 0.762±0.079 d | 7.156±0.523 c | 0.064±0.004 c |
Figure 2 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on kale seedling growth at 5 days post-germination under salt stress Control-1, Control-2, SA and AIP are the same as Table 2.
Figure 3 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on SA content (A), PAL activity (B) and BoPAL1-2 expression (C, D) in kale seeds during germination under salt stress PAL: Phenylalanine ammonia-lyase. Control-1, Control-2, SA and AIP are the same as Table 2. Different lowercase letters above the bars indicate significant differences (P<0.05) among treatments at the same sown time.
| Germination time (h) | Treatments | Ion content (mg·g-1 FW) | |||
|---|---|---|---|---|---|
| Na+ | K+ | Ca2+ | Mg2+ | ||
| 12 | Control-1 | 2.553±0.219 c | 1.471±0.134 a | 0.221±0.018 a | 0.431±0.026 a |
| Control-2 | 3.711±0.431 a | 1.227±0.203 b | 0.192±0.012 b | 0.313±0.033 c | |
| SA | 3.292±0.331 b | 1.435±0.053 a | 0.183±0.009 b | 0.372±0.042 b | |
| AIP | 3.809±0.504 a | 1.170±0.077 c | 0.211±0.018 a | 0.211±0.018 d | |
| 24 | Control-1 | 3.951±0.412 c | 1.682±0.117 a | 0.148±0.017 a | 0.477±0.033 a |
| Control-2 | 5.587±0.581 a | 1.427±0.109 b | 0.123±0.017 b | 0.385±0.049 b | |
| SA | 5.108±0.432 b | 1.623±0.152 a | 0.112±0.006 b | 0.462±0.053 a | |
| AIP | 5.717±0.367 a | 1.301±0.083 c | 0.118±0.008 b | 0.321±0.019 c | |
| 36 | Control-1 | 3.762±0.319 c | 1.677±0.092 a | 0.176±0.074 a | 0.597±0.055 a |
| Control-2 | 6.193±0.419 a | 1.486±0.106 c | 0.093±0.012 b | 0.511±0.022 b | |
| SA | 5.654±0.384 b | 1.583±0.201 b | 0.101±0.004 b | 0.590±0.044 a | |
| AIP | 6.202±0.609 a | 1.472±0.013 c | 0.095±0.011 b | 0.367±0.048 c | |
| 48 | Control-1 | 4.188±0.381 b | 1.769±0.208 a | 0.132±0.008 a | 0.646±0.054 a |
| Control-2 | 5.772±0.720 a | 1.311±0.011 c | 0.073±0.003 c | 0.491±0.060 c | |
| SA | 5.790±0.364 a | 1.422±0.015 b | 0.072±0.009 c | 0.582±0.015 b | |
| AIP | 5.831±0.519 a | 1.226±0.064 d | 0.094±0.014 b | 0.401±0.008 d | |
Table 4 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on ions contents during seed germination of kale under salt stress
| Germination time (h) | Treatments | Ion content (mg·g-1 FW) | |||
|---|---|---|---|---|---|
| Na+ | K+ | Ca2+ | Mg2+ | ||
| 12 | Control-1 | 2.553±0.219 c | 1.471±0.134 a | 0.221±0.018 a | 0.431±0.026 a |
| Control-2 | 3.711±0.431 a | 1.227±0.203 b | 0.192±0.012 b | 0.313±0.033 c | |
| SA | 3.292±0.331 b | 1.435±0.053 a | 0.183±0.009 b | 0.372±0.042 b | |
| AIP | 3.809±0.504 a | 1.170±0.077 c | 0.211±0.018 a | 0.211±0.018 d | |
| 24 | Control-1 | 3.951±0.412 c | 1.682±0.117 a | 0.148±0.017 a | 0.477±0.033 a |
| Control-2 | 5.587±0.581 a | 1.427±0.109 b | 0.123±0.017 b | 0.385±0.049 b | |
| SA | 5.108±0.432 b | 1.623±0.152 a | 0.112±0.006 b | 0.462±0.053 a | |
| AIP | 5.717±0.367 a | 1.301±0.083 c | 0.118±0.008 b | 0.321±0.019 c | |
| 36 | Control-1 | 3.762±0.319 c | 1.677±0.092 a | 0.176±0.074 a | 0.597±0.055 a |
| Control-2 | 6.193±0.419 a | 1.486±0.106 c | 0.093±0.012 b | 0.511±0.022 b | |
| SA | 5.654±0.384 b | 1.583±0.201 b | 0.101±0.004 b | 0.590±0.044 a | |
| AIP | 6.202±0.609 a | 1.472±0.013 c | 0.095±0.011 b | 0.367±0.048 c | |
| 48 | Control-1 | 4.188±0.381 b | 1.769±0.208 a | 0.132±0.008 a | 0.646±0.054 a |
| Control-2 | 5.772±0.720 a | 1.311±0.011 c | 0.073±0.003 c | 0.491±0.060 c | |
| SA | 5.790±0.364 a | 1.422±0.015 b | 0.072±0.009 c | 0.582±0.015 b | |
| AIP | 5.831±0.519 a | 1.226±0.064 d | 0.094±0.014 b | 0.401±0.008 d | |
| Germination time (h) | Treatments | Osmotic adjustment substance | ||
|---|---|---|---|---|
| Soluble sugar (mg·g-1 FW) | Soluble protein (mg·g-1 FW) | Free proline (μg·g-1 FW) | ||
| 12 | Control-1 | 13.56±0.81 d | 3.57±0.40 d | 108.19±1.23 c |
| Control-2 | 23.19±1.73 b | 8.11±0.71 b | 143.77±1.35 b | |
| SA | 29.28±2.00 a | 12.17±1.03 a | 156.53±1.17 a | |
| AIP | 19.13±1.23 c | 6.59±0.33 c | 157.11±0.90 a | |
| 24 | Control-1 | 17.11±0.92 c | 6.17±0.55 c | 115.64±0.87 c |
| Control-2 | 26.35±1.74 b | 9.27±0.49 b | 167.59±2.11 a | |
| SA | 33.27±4.02 a | 13.56±0.84 a | 142.13±1.33 b | |
| AIP | 25.77±3.19 b | 6.59±0.30 c | 122.64±2.16 c | |
| 36 | Control-1 | 23.85±1.39 d | 6.12±0.41 c | 117.26±1.55 c |
| Control-2 | 37.50±2.47 b | 11.73±1.23 a | 141.33±1.46 b | |
| SA | 43.59±4.33 a | 12.19±1.19 a | 172.57±1.15 a | |
| AIP | 30.66±2.20 c | 9.13±0.52 b | 109.33±0.82 c | |
| 48 | Control-1 | 22.74±3.10 c | 5.80±0.37 b | 133.52±0.81 c |
| Control-2 | 34.19±2.79 a | 8.62±0.79 a | 167.29±1.31 b | |
| SA | 36.82±2.12 a | 8.93±0.64 a | 128.19±1.55 c | |
| AIP | 26.70±2.34 b | 6.18±0.56 b | 186.53±2.08 a | |
Table 5 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on osmotic adjustment substances during seed germination of kale under salt stress
| Germination time (h) | Treatments | Osmotic adjustment substance | ||
|---|---|---|---|---|
| Soluble sugar (mg·g-1 FW) | Soluble protein (mg·g-1 FW) | Free proline (μg·g-1 FW) | ||
| 12 | Control-1 | 13.56±0.81 d | 3.57±0.40 d | 108.19±1.23 c |
| Control-2 | 23.19±1.73 b | 8.11±0.71 b | 143.77±1.35 b | |
| SA | 29.28±2.00 a | 12.17±1.03 a | 156.53±1.17 a | |
| AIP | 19.13±1.23 c | 6.59±0.33 c | 157.11±0.90 a | |
| 24 | Control-1 | 17.11±0.92 c | 6.17±0.55 c | 115.64±0.87 c |
| Control-2 | 26.35±1.74 b | 9.27±0.49 b | 167.59±2.11 a | |
| SA | 33.27±4.02 a | 13.56±0.84 a | 142.13±1.33 b | |
| AIP | 25.77±3.19 b | 6.59±0.30 c | 122.64±2.16 c | |
| 36 | Control-1 | 23.85±1.39 d | 6.12±0.41 c | 117.26±1.55 c |
| Control-2 | 37.50±2.47 b | 11.73±1.23 a | 141.33±1.46 b | |
| SA | 43.59±4.33 a | 12.19±1.19 a | 172.57±1.15 a | |
| AIP | 30.66±2.20 c | 9.13±0.52 b | 109.33±0.82 c | |
| 48 | Control-1 | 22.74±3.10 c | 5.80±0.37 b | 133.52±0.81 c |
| Control-2 | 34.19±2.79 a | 8.62±0.79 a | 167.29±1.31 b | |
| SA | 36.82±2.12 a | 8.93±0.64 a | 128.19±1.55 c | |
| AIP | 26.70±2.34 b | 6.18±0.56 b | 186.53±2.08 a | |
Figure 4 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on H2O2 (A) and O2-. (B) contents of kale seed during germination under salt stress Control-1, Control-2, SA and AIP are the same as Table 2. Different lowercase letters above the bars indicate significant differences (P<0.05) among treatments at the same sown time.
Figure 5 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on the activities of catalase (CAT) (A), superoxide dismutase (SOD) (B), and peroxidase (POD) (C) in kale seeds during germination under salt stress Control-1, Control-2, SA and AIP are the same as Table 2.
Figure 6 Effects of salicylic acid (SA) and SA synthesis inhibitor (2-aminoindano-2-phosphonic acid, AIP) on gene expression of BoCAT1 (A), BoCAT2 (B), BoSOD1 (C), BoSOD3 (D), BoPOD2 (E), and BoPOD3 (F) during seed germination of kale under salt stress Control-1, Control-2, SA and AIP are the same as Table 2. Different lowercase letters above the bars indicate significant differences (P<0.05) among treatments at the same sown time.
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