Salinity Depression in Vitro Impress the Growth Potential and, Mediates the Antioxidants Pool and the Tolerance Genes Expression of Grapevine
Undoubtedly, salinity is the major environmental stress affecting the crops growth, development and yield. Grapevine is a dominant horticulture crop and mildly sensitive to the salinity. Salinity triggers variations at the cellular and molecular levels and hence induces some specific genes expression. The responses of two grapevine cultivars (Fakhri and Sultanin) were evaluated for its tolerance to the salinity stress in vitro. The results for the explants exposed to the salinity revealed that the viability, fresh weight of the regenerated explant and the proliferation rate were declined compared to the control ones. The activity of SOD enzyme and MDA content were increased with salinity. However, protein content declined. There was no significant difference in CAT and APX activities with the salinity treatment. With salinity adding up, the DREB/CBFs genes expression pattern was significantly increased in both cultivars. ‘Fakhri’ was more responsive in growth parameters and the activity of antioxidant enzymes and higher expression rate of DREB/CBFs under salinity compared to the variety ‘Sultanin’.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Table 1. Reaction mixture for cDNA synthesis
Volume |
Reactive |
0.5 µl |
Vivantis RT Enzyme Mix |
2 µl |
Buffer RT Enzyme |
0.5 µl |
Oligo RT Primer (50 µM) |
0.5 µl |
Random 6 mers (100 µM) |
1 µl |
dNTP |
11.5 µl |
DDW |
5 µl |
Total RNA (500 ng) |
20 µl |
Total |
Due to technical limitations, table 2 is only available as a download in the Supplemental Files section.
Table 3. The composition of reaction mixture for RT-RCR
Volume |
Reactive |
2 µl |
RT reaction solution (cDNA) |
0.4 µl |
Primer F |
0.4 µl |
Primer R |
10 µl |
Power SYBR Green PCR Master Mix |
7.2 µl |
DDW |
20 µl |
Total |
Table 4. The effects of NaCl treatments on some physiological traits in two grape cultivars
Cultivar |
NaCl(gL-1) |
Shoot length (mm) |
Shoot weight (mg) |
Viability (%) |
Number of shoots |
Proliferation ratio |
Number of leaves/shoots |
Number of 3-node shoots |
0 |
35.13±0.99a |
324.79±41.66a |
100±0a |
6±0.61a |
14.25±3.36a |
4.5±0.25a |
2.75±0.21a |
|
0.25 |
23.62±0.77b |
267.04±17.87ab |
87.50±2.16b |
5±0.35a |
8.75±0.41bc |
3±0.35bcde |
2.25±0.21ab |
|
Sultanin |
0.5 |
16.33±0.81c |
182.12±31.72bc |
86.25±3.75b |
4.25±0.89ab |
5.5±0.55cde |
2.75±0.21cdef |
1.75±0.41abcd |
0.75 |
10.18±0.45d |
93.05±6.82cd |
76.25±2.72c |
2.25±0.41bc |
3±0.61de |
2.25±0.21ef |
1.5±0.25abcd |
|
1 |
7.93±0.34d |
27.59±4.64d |
21.25±3.69e |
1.5±0.43c |
1.75±0.41e |
1.75±0.41f |
0.5±0.25d |
|
0 |
33.38±3.25a |
314.72±16.14a |
100±0a |
6.25±0.96a |
11.75±2.21ab |
4±0.35ab |
2.75±0.21a |
|
0.25 |
22.08±0.59b |
294.67±70.3ab |
100±0a |
4.5±0.55ab |
7.5±0.55bcd |
3.75±0.21abc |
2.75±0.21a |
|
Fakhri |
0.5 |
16.49±0.71c |
263.74±37.74ab |
100±0a |
4.25±0.96ab |
5.25±0.21cde |
3.5±0.25abcd |
2±0.5abc |
0.75 |
11.96±0.52d |
105.86±11.09cd |
83.75±3.69bc |
2.75±0.41bc |
2.75±0.21de |
2.75±0.41cdef |
1.25±0.54bcd |
|
1 |
9.10±0.64d |
49.92±11.01d |
33.75±2.72d |
2.25±0.21bc |
1.25±0.21e |
2.5±0.25def |
0.75±0.41cd |
Means by the same latter within each column are not significantly different at 0.05 level, according to the Duncan's multiple range test. Data are mean±SD (n=4 replicates)
Table 5. Coefficients of correlation values for the pairs of studied characters in two grape cultivars under NaCl
|
Y1 (Shoot length) |
Y2 (Shoot weight) |
Y3 (Viability) |
Y4 (Number of shoots) |
Y5 (Proliferation ratio) |
Y6 (Number of leaves/shoots) |
Y7 (Number of 3-node shoots) |
Y8 (APX) |
Y9 (GPX) |
Y10 (CAT) |
Y11 (SOD) |
Y12 (MDA) |
Y13 (Protein) |
Y1 |
1 |
|
|
|
|
|
|
|
|
|
|
|
|
Y2 |
0.769** |
1 |
|
|
|
|
|
|
|
|
|
|
|
Y3 |
0.670** |
0.734** |
1 |
|
|
|
|
|
|
|
|
|
|
Y4 |
0.766** |
0.742** |
0.626** |
1 |
|
|
|
|
|
|
|
|
|
Y5 |
0.841** |
0.728** |
0.575** |
0.692** |
1 |
|
|
|
|
|
|
|
|
Y6 |
0.711** |
0.689** |
0.617** |
0.544** |
0.591** |
1 |
|
|
|
|
|
|
|
Y7 |
0.718** |
0.760** |
0.633** |
0.597** |
0.545** |
0.652** |
1 |
|
|
|
|
|
|
Y8 |
0.033 |
-0.047 |
-0.207 |
-0.016 |
0.090 |
-0.025 |
-0.071 |
1 |
|
|
|
|
|
Y9 |
0.102 |
-0.013 |
-0.041 |
-0.035 |
0.178 |
-0.057 |
0.043 |
0.466** |
1 |
|
|
|
|
Y10 |
-0.053 |
-0.135 |
-0.114 |
-0.058 |
0.040 |
-0.126 |
0.017 |
0.679** |
0.399* |
1 |
|
|
|
Y11 |
0.076 |
0.098 |
0.126 |
-0.028 |
0.215 |
-0.022 |
0.197 |
0.302 |
0.507** |
0.261 |
1 |
|
|
Y12 |
0.134 |
0.074 |
-0.106 |
0.272 |
0.038 |
0.121 |
0.188 |
-0.272 |
-0.377* |
-0.253 |
-0.328* |
1 |
|
Y13 |
0.376* |
0.098 |
0.065 |
0.130 |
0.377* |
0.128 |
0.132 |
0.230 |
0.025 |
0.116 |
0.278 |
-0.046 |
1 |
**, * significant at p≤0.01 probability and significant at p≤0.05 probability, respectively.
Table 6. Total variance explained |
|||
|
Initial Eigenvalues |
||
Components |
Total |
% of Variance |
Cumulative % |
1 |
5.172 |
39.783 |
39.783 |
2 |
2.680 |
20.612 |
60.395 |
3 |
1.132 |
8.710 |
69.105 |
4 |
0.991 |
7.620 |
76.725 |
5 |
0.739 |
5.688 |
82.412 |
6 |
0.587 |
4.512 |
86.925 |
7 |
0.483 |
3.716 |
90.641 |
8 |
0.358 |
2.753 |
93.394 |
9 |
0.273 |
2.102 |
95.496 |
10 |
0.243 |
1.872 |
97.368 |
11 |
0.156 |
1.201 |
98.570 |
12 |
0.123 |
0.946 |
99.516 |
13 |
0.063 |
0.484 |
100.000 |
Extraction Method: Principal Component Analysis. |
Table 7. Total variance explained |
||
Component Matrixa |
||
|
1 |
2 |
Y1 |
0.930 |
0.060 |
Y2 |
0.905 |
-0.053 |
Y3 |
0.847 |
0.198 |
Y4 |
0.832 |
-0.097 |
Y5 |
0.823 |
0.002 |
Y6 |
0.802 |
-0.070 |
Y7 |
0.797 |
-0.099 |
Y8 |
-0.040 |
0.788 |
Y9 |
0.032 |
0.758 |
Y10 |
-0.083 |
0.733 |
Y11 |
0.138 |
0.672 |
Y12 |
0.139 |
-0.560 |
Y13 |
0.274 |
0.334 |
Extraction Method: Principal Component Analysis (PCA). 1 (PCA1) and 2 (PCA2) a: 3 components extracted. Y1 (Shoot length),Y2 (Shoot weight),Y3 (Viability),Y4 (Number of shoots),Y5 (Proliferation ratio),Y6 (Number of leaves/shoots),Y7 (Number of 3-node shoots),Y8 (APX),Y9 (GPX),Y10 (CAT), Y11 (SOD),Y12 (MDA),Y13 (Protein) |
This is a list of supplementary files associated with this preprint. Click to download.
Posted 21 Dec, 2020
Invitations sent on 14 Feb, 2021
On 06 Jan, 2021
On 16 Dec, 2020
On 16 Dec, 2020
On 15 Dec, 2020
Salinity Depression in Vitro Impress the Growth Potential and, Mediates the Antioxidants Pool and the Tolerance Genes Expression of Grapevine
Posted 21 Dec, 2020
Invitations sent on 14 Feb, 2021
On 06 Jan, 2021
On 16 Dec, 2020
On 16 Dec, 2020
On 15 Dec, 2020
Undoubtedly, salinity is the major environmental stress affecting the crops growth, development and yield. Grapevine is a dominant horticulture crop and mildly sensitive to the salinity. Salinity triggers variations at the cellular and molecular levels and hence induces some specific genes expression. The responses of two grapevine cultivars (Fakhri and Sultanin) were evaluated for its tolerance to the salinity stress in vitro. The results for the explants exposed to the salinity revealed that the viability, fresh weight of the regenerated explant and the proliferation rate were declined compared to the control ones. The activity of SOD enzyme and MDA content were increased with salinity. However, protein content declined. There was no significant difference in CAT and APX activities with the salinity treatment. With salinity adding up, the DREB/CBFs genes expression pattern was significantly increased in both cultivars. ‘Fakhri’ was more responsive in growth parameters and the activity of antioxidant enzymes and higher expression rate of DREB/CBFs under salinity compared to the variety ‘Sultanin’.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Table 1. Reaction mixture for cDNA synthesis
Volume |
Reactive |
0.5 µl |
Vivantis RT Enzyme Mix |
2 µl |
Buffer RT Enzyme |
0.5 µl |
Oligo RT Primer (50 µM) |
0.5 µl |
Random 6 mers (100 µM) |
1 µl |
dNTP |
11.5 µl |
DDW |
5 µl |
Total RNA (500 ng) |
20 µl |
Total |
Due to technical limitations, table 2 is only available as a download in the Supplemental Files section.
Table 3. The composition of reaction mixture for RT-RCR
Volume |
Reactive |
2 µl |
RT reaction solution (cDNA) |
0.4 µl |
Primer F |
0.4 µl |
Primer R |
10 µl |
Power SYBR Green PCR Master Mix |
7.2 µl |
DDW |
20 µl |
Total |
Table 4. The effects of NaCl treatments on some physiological traits in two grape cultivars
Cultivar |
NaCl(gL-1) |
Shoot length (mm) |
Shoot weight (mg) |
Viability (%) |
Number of shoots |
Proliferation ratio |
Number of leaves/shoots |
Number of 3-node shoots |
0 |
35.13±0.99a |
324.79±41.66a |
100±0a |
6±0.61a |
14.25±3.36a |
4.5±0.25a |
2.75±0.21a |
|
0.25 |
23.62±0.77b |
267.04±17.87ab |
87.50±2.16b |
5±0.35a |
8.75±0.41bc |
3±0.35bcde |
2.25±0.21ab |
|
Sultanin |
0.5 |
16.33±0.81c |
182.12±31.72bc |
86.25±3.75b |
4.25±0.89ab |
5.5±0.55cde |
2.75±0.21cdef |
1.75±0.41abcd |
0.75 |
10.18±0.45d |
93.05±6.82cd |
76.25±2.72c |
2.25±0.41bc |
3±0.61de |
2.25±0.21ef |
1.5±0.25abcd |
|
1 |
7.93±0.34d |
27.59±4.64d |
21.25±3.69e |
1.5±0.43c |
1.75±0.41e |
1.75±0.41f |
0.5±0.25d |
|
0 |
33.38±3.25a |
314.72±16.14a |
100±0a |
6.25±0.96a |
11.75±2.21ab |
4±0.35ab |
2.75±0.21a |
|
0.25 |
22.08±0.59b |
294.67±70.3ab |
100±0a |
4.5±0.55ab |
7.5±0.55bcd |
3.75±0.21abc |
2.75±0.21a |
|
Fakhri |
0.5 |
16.49±0.71c |
263.74±37.74ab |
100±0a |
4.25±0.96ab |
5.25±0.21cde |
3.5±0.25abcd |
2±0.5abc |
0.75 |
11.96±0.52d |
105.86±11.09cd |
83.75±3.69bc |
2.75±0.41bc |
2.75±0.21de |
2.75±0.41cdef |
1.25±0.54bcd |
|
1 |
9.10±0.64d |
49.92±11.01d |
33.75±2.72d |
2.25±0.21bc |
1.25±0.21e |
2.5±0.25def |
0.75±0.41cd |
Means by the same latter within each column are not significantly different at 0.05 level, according to the Duncan's multiple range test. Data are mean±SD (n=4 replicates)
Table 5. Coefficients of correlation values for the pairs of studied characters in two grape cultivars under NaCl
|
Y1 (Shoot length) |
Y2 (Shoot weight) |
Y3 (Viability) |
Y4 (Number of shoots) |
Y5 (Proliferation ratio) |
Y6 (Number of leaves/shoots) |
Y7 (Number of 3-node shoots) |
Y8 (APX) |
Y9 (GPX) |
Y10 (CAT) |
Y11 (SOD) |
Y12 (MDA) |
Y13 (Protein) |
Y1 |
1 |
|
|
|
|
|
|
|
|
|
|
|
|
Y2 |
0.769** |
1 |
|
|
|
|
|
|
|
|
|
|
|
Y3 |
0.670** |
0.734** |
1 |
|
|
|
|
|
|
|
|
|
|
Y4 |
0.766** |
0.742** |
0.626** |
1 |
|
|
|
|
|
|
|
|
|
Y5 |
0.841** |
0.728** |
0.575** |
0.692** |
1 |
|
|
|
|
|
|
|
|
Y6 |
0.711** |
0.689** |
0.617** |
0.544** |
0.591** |
1 |
|
|
|
|
|
|
|
Y7 |
0.718** |
0.760** |
0.633** |
0.597** |
0.545** |
0.652** |
1 |
|
|
|
|
|
|
Y8 |
0.033 |
-0.047 |
-0.207 |
-0.016 |
0.090 |
-0.025 |
-0.071 |
1 |
|
|
|
|
|
Y9 |
0.102 |
-0.013 |
-0.041 |
-0.035 |
0.178 |
-0.057 |
0.043 |
0.466** |
1 |
|
|
|
|
Y10 |
-0.053 |
-0.135 |
-0.114 |
-0.058 |
0.040 |
-0.126 |
0.017 |
0.679** |
0.399* |
1 |
|
|
|
Y11 |
0.076 |
0.098 |
0.126 |
-0.028 |
0.215 |
-0.022 |
0.197 |
0.302 |
0.507** |
0.261 |
1 |
|
|
Y12 |
0.134 |
0.074 |
-0.106 |
0.272 |
0.038 |
0.121 |
0.188 |
-0.272 |
-0.377* |
-0.253 |
-0.328* |
1 |
|
Y13 |
0.376* |
0.098 |
0.065 |
0.130 |
0.377* |
0.128 |
0.132 |
0.230 |
0.025 |
0.116 |
0.278 |
-0.046 |
1 |
**, * significant at p≤0.01 probability and significant at p≤0.05 probability, respectively.
Table 6. Total variance explained |
|||
|
Initial Eigenvalues |
||
Components |
Total |
% of Variance |
Cumulative % |
1 |
5.172 |
39.783 |
39.783 |
2 |
2.680 |
20.612 |
60.395 |
3 |
1.132 |
8.710 |
69.105 |
4 |
0.991 |
7.620 |
76.725 |
5 |
0.739 |
5.688 |
82.412 |
6 |
0.587 |
4.512 |
86.925 |
7 |
0.483 |
3.716 |
90.641 |
8 |
0.358 |
2.753 |
93.394 |
9 |
0.273 |
2.102 |
95.496 |
10 |
0.243 |
1.872 |
97.368 |
11 |
0.156 |
1.201 |
98.570 |
12 |
0.123 |
0.946 |
99.516 |
13 |
0.063 |
0.484 |
100.000 |
Extraction Method: Principal Component Analysis. |
Table 7. Total variance explained |
||
Component Matrixa |
||
|
1 |
2 |
Y1 |
0.930 |
0.060 |
Y2 |
0.905 |
-0.053 |
Y3 |
0.847 |
0.198 |
Y4 |
0.832 |
-0.097 |
Y5 |
0.823 |
0.002 |
Y6 |
0.802 |
-0.070 |
Y7 |
0.797 |
-0.099 |
Y8 |
-0.040 |
0.788 |
Y9 |
0.032 |
0.758 |
Y10 |
-0.083 |
0.733 |
Y11 |
0.138 |
0.672 |
Y12 |
0.139 |
-0.560 |
Y13 |
0.274 |
0.334 |
Extraction Method: Principal Component Analysis (PCA). 1 (PCA1) and 2 (PCA2) a: 3 components extracted. Y1 (Shoot length),Y2 (Shoot weight),Y3 (Viability),Y4 (Number of shoots),Y5 (Proliferation ratio),Y6 (Number of leaves/shoots),Y7 (Number of 3-node shoots),Y8 (APX),Y9 (GPX),Y10 (CAT), Y11 (SOD),Y12 (MDA),Y13 (Protein) |