Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (22): 4027-4038.doi: 10.3864/j.issn.0578-1752.2019.22.008

• GERMPLASM RESOURCES • Previous Articles     Next Articles

Screening for Germplasms Tolerant to Salt at Germination Stage and Response of Protective Enzymes to Salt Stress in Foxtail Millet

QIN Ling,ZHANG YanTing,CHEN ErYing,YANG YanBing,LI FeiFei,GUAN YanAn()   

  1. Crop Research Institute, Shandong Academy of Agricultural Sciences/Featured Crops Engineering Laboratory of Shandong Province, Jinan 250100
  • Received:2019-06-05 Accepted:2019-07-08 Online:2019-11-16 Published:2019-11-16
  • Contact: YanAn GUAN E-mail:yguan65@163.com

Abstract:

【Objective】The purpose of this study was to select germplasms tolerant to salt at germination stage, to establish an evaluation criterion for salt tolerance, and to investigate the effect of salt stress on protective enzyme system of different genotypes of foxtail millet at seedling stage. The results could propose an identification method and excellent genetic resources for research on salt-tolerant mechanisms of foxtail millet.【Method】54 foxtail millet varieties from different eco-regions were applied as experimental materials. Salt stress was implemented by adding 1.5% NaCl solution, and distilled water was used as the control. A petri dish based germination method was used to assess the salt tolerance of foxtail millet varieties in artificial climate incubator. Six salt-tolerant indexes were investigated in each treatment, including relative germination potential, relative germination rate, relative coleoptile length, relative radical root length, relative radical root / coleoptiles, and relative salt damage rate. A comprehensive analysis of the salt tolerance of different varieties was conducted via correlation analysis, principal component analysis and clustering analysis. Furthermore, pot experiment was conducted to cultivate selected varieties with different salt tolerance under both salt stress and normal conditions, and superoxide dismutase (SOD) activity, peroxidase (POD) activity, and catalase (CAT) activity in shoot were analyzed. And then the differences of physiological responses between those genotypes under salt stress were analyzed.【Result】Under salt stress of 1.5% NaCl , a significant positive correlation was detected between relative germination potential and relative germination(r=0.51, P<0.01) and relative coleoptile length (r=0.54, P<0.01). There was a significant positive correlation between relative coleoptile length and relative radical root length (r=0.64, P<0.01), while the relative salt damage rate showed a significant negative correlation with relative germination rate (r=-0.37, P<0.01) and relative coleoptile length (r=-0.51, P<0.01). Principal component analysis transformed the six single indexes in the salt-stressed germination stage of foxtail millet to three independent comprehensive indexes. Subordinate function analysis was conducted to obtain the comprehensive assessment value (D-value) of salt tolerance of different varieties at germination stage. 54 foxtail millet varieties were classified into five salt tolerant types based on clustering analysis, including extremely salt sensitive, salt sensitive, moderate salt tolerance, salt tolerance and high salt tolerance. The varieties with high salt tolerance were Yangu 13, Longgu 3 from spring sowing region of northwest China, and Jigu 16, Ai88 from summer sowing region of north China. The activities of SOD, POD, and CAT were increased and then decreased under salt stress condition. SOD, POD, and CAT in shoot increased in response to salt stress, and the activity of those enzymes in Jigu 16 was significantly higher than that in Lugu 1.【Conclusion】Remarkable variations in salt tolerance were detected among the 54 genotypes of foxtail millet. The systematical and clear results can be obtained by membership function analysis, and the salt tolerance of foxtail millet can be evaluated objectively and comprehensively. The leaf protective enzymes are important for the adaptability of foxtail millet to salt stress in seedling stage.

Key words: foxtail millet (Setaria italica L.), salt-stress, germination stage, comprehensive evaluation, protective enzymes

Table 1

Coefficient and variation of each character of different millet varieties under salt stress"

指标
Index
变化范围
Variation range
平均值
Mean
变异系数
Coefficient of variance
相对发芽势 Relative germination potential(%) 0.00—74.57 14.24 100.27
相对发芽率 Relative germination rate(%) 3.81—93.77 36.15 56.96
相对胚芽长 Relative coleoptile length(%) 8.66—59.48 24.45 36.33
相对胚根长 Relative radical root length(%) 4.52—33.62 9.83 47.64
相对根芽比 Relative radical root / coleoptiles(%) 19.83—105.46 41.67 35.88
相对盐害率 Relative salt damage rate(%) 6.33—96.19 63.85 32.25

Table 2

Correlation analysis of coefficient for each character of different foxtail millet varieties under salt stress"

相关系数
Correlation coefficient
相对发芽势
Relative germination potential (X1)
相对发芽率
Relative germination rate (X2)
相对胚芽长
Relative coleoptile length (X3)
相对胚根长
Relative radical root length (X4)
相对根芽比
Relative radical root / coleoptiles (X5)
相对盐害率
Relative salt damage rate (X6)
X1 1
X2 0.51** 1
X3 0.54** 0.46** 1
X4 0.05 0.10 0.64** 1
X5 -0.37** -0.28** -0.27** 0.47** 1
X6 -0.51** -1.00** -0.46** -0.10 0.28** 1

Table 3

Eigenvalue and contribution and eigenvectors of 3 comprehensive indexes [CI(x)]"

项目 Item CI1 CI2 CI3
特征值Eigen value 2.984 1.613 0.870
贡献率Contributive ratio (%) 49.73 26.89 14.50
累积贡献率
Cumulative contributive ratio (%)
49.73 76.62 91.12
特征向量Eigenvector X1 0.435 -0.092 0.324
特征向量Eigenvector X2 0.517 -0.075 -0.465
特征向量Eigenvector X3 0.438 0.340 0.462
特征向量Eigenvector X4 0.144 0.748 0.099
特征向量Eigenvector X5 -0.252 0.552 -0.490
特征向量Eigenvector X6 -0.517 0.075 0.465

Table 4

Comprehensive index(CIx), index weight, μ (X) and D value of each variety"

序号
Number
CI1 CI2 CI3 μ (X1) μ (X2) μ (X3) D 综合评价
Comprehensive valuation
1 -2.2796 -0.5447 0.4050 0.0955 0.1295 0.5460 0.1772 高度盐敏感Extremely salt sensitivity
2 1.3460 0.2178 -0.1585 0.4899 0.2264 0.4308 0.4028 中度耐盐Moderate salt tolerance
3 1.2316 0.2696 -1.8077 0.4775 0.2330 0.0939 0.3443 中度耐盐Moderate salt tolerance
4 0.0000 0.8496 -0.0200 0.3435 0.3067 0.4591 0.3510 中度耐盐Moderate salt tolerance
5 -0.0623 -1.2831 0.4707 0.3367 0.0357 0.5594 0.2833 盐敏感Salt sensitivity
6 2.2398 3.5566 0.6492 0.5872 0.6506 0.5958 0.6073 高度耐盐High salt tolerance
7 -0.4241 -0.1059 -0.6293 0.2973 0.1853 0.3346 0.2702 盐敏感Salt sensitivity
8 -0.5756 1.9376 2.6273 0.2809 0.4449 1.0000 0.4437 耐盐Salt tolerance
9 -0.3235 -0.5754 -0.4567 0.3083 0.1256 0.3699 0.2642 盐敏感Salt sensitivity
10 -1.0946 0.0800 -0.5070 0.2244 0.2089 0.3596 0.2413 盐敏感Salt sensitivity
11 -0.1767 0.4078 -1.9838 0.3243 0.2505 0.0579 0.2601 盐敏感Salt sensitivity
12 1.5976 -0.0443 -1.1515 0.5173 0.1931 0.2279 0.3756 中度耐盐Moderate salt tolerance
13 0.6795 -0.2044 -0.7963 0.4174 0.1727 0.3005 0.3266 中度耐盐Moderate salt tolerance
14 -0.4494 -1.1064 1.4951 0.2946 0.0581 0.7687 0.3002 盐敏感Salt sensitivity
15 2.0224 0.6177 0.1566 0.5635 0.2772 0.4952 0.4682 耐盐Salt tolerance
16 1.7565 -1.1427 0.0451 0.5346 0.0535 0.4724 0.3827 中度耐盐Moderate salt tolerance
17 0.9306 -0.6810 -0.2228 0.4447 0.1122 0.4177 0.3423 中度耐盐Moderate salt tolerance
18 -1.4052 -0.1881 0.5329 0.1906 0.1748 0.5721 0.2466 盐敏感Salt sensitivity
19 -0.7363 6.3065 0.4175 0.2634 1.0000 0.5485 0.5261 高度耐盐High salt tolerance
20 0.4163 -0.8748 0.1120 0.3888 0.0876 0.4861 0.3154 中度耐盐Moderate salt tolerance
21 2.619 -1.4495 -2.1504 0.6284 0.0145 0.0238 0.3511 中度耐盐Moderate salt tolerance
22 -0.0984 -0.9923 0.3280 0.3328 0.0726 0.5302 0.2874 盐敏感Salt sensitivity
23 2.0215 -0.3289 0.3369 0.5634 0.1569 0.5320 0.4385 耐盐 Salt tolerance
24 0.8321 -1.5640 0.2352 0.4340 0.0000 0.5113 0.3182 中度耐盐Moderate salt tolerance
25 -0.3626 -0.8436 -0.2953 0.3040 0.0915 0.4029 0.2570 盐敏感Salt sensitivity
26 -0.6926 -1.2175 0.2139 0.2681 0.0440 0.5069 0.2400 盐敏感Salt sensitivity
27 -1.9623 -0.6357 -0.0949 0.1300 0.1179 0.4438 0.1764 高度盐敏感Extremely salt sensitivity
28 0.4516 1.1999 -0.8049 0.3926 0.3512 0.2987 0.3655 中度耐盐Moderate salt tolerance
29 1.2783 -0.4670 0.5348 0.4826 0.1394 0.5725 0.3956 中度耐盐Moderate salt tolerance
30 -1.1852 -0.1785 0.5406 0.2145 0.1760 0.5737 0.2603 盐敏感Salt sensitivity
31 -3.1570 1.8624 -2.2671 0.0000 0.4353 0.0000 0.1285 高度盐敏感Extremely salt sensitivity
32 -3.0920 -0.8026 -0.0456 0.0071 0.0967 0.4539 0.1046 高度盐敏感Extremely salt sensitivity
33 -1.5469 -1.1680 0.6173 0.1752 0.0503 0.5893 0.2042 高度盐敏感Extremely salt sensitivity
34 -1.8683 -1.2398 1.1237 0.1402 0.0412 0.6928 0.1989 高度盐敏感Extremely salt sensitivity
35 1.0959 1.4691 -0.4824 0.4627 0.3854 0.3646 0.4243 耐盐Salt tolerance
36 -0.5245 -0.4524 0.4839 0.2864 0.1412 0.5621 0.2874 盐敏感Salt sensitivity
37 -1.9564 -0.5711 0.2578 0.1306 0.1262 0.5159 0.1906 高度盐敏感Extremely salt sensitivity
38 5.6668 -1.3401 0.1074 0.9600 0.0284 0.4851 0.6096 高度耐盐High salt tolerance
39 -0.4107 -0.5841 1.7251 0.2988 0.1245 0.8157 0.3296 中度耐盐Moderate salt tolerance
40 -0.0538 -1.0250 0.4973 0.3376 0.0685 0.5648 0.2944 盐敏感Salt sensitivity
41 6.0342 1.7618 1.6937 1.0000 0.4226 0.8093 0.7993 高度耐盐High salt tolerance
42 -2.1428 -0.5681 0.0462 0.1103 0.1265 0.4726 0.1728 高度盐敏感Extremely salt sensitivity
43 0.4245 0.1161 0.4252 0.3897 0.2135 0.5501 0.3632 中度耐盐Moderate salt tolerance
44 -1.1984 0.6045 0.4779 0.2131 0.2755 0.5608 0.2868 盐敏感Salt sensitivity
45 -0.9893 -0.9606 0.5596 0.2358 0.0767 0.5775 0.2432 盐敏感Salt sensitivity
46 -2.2146 -0.6243 -0.9891 0.1025 0.1194 0.2611 0.1327 高度盐敏感Extremely salt sensitivity
47 0.5139 1.8569 0.6272 0.3994 0.4346 0.5913 0.4403 耐盐Salt-tolerance
48 1.3441 0.245 -0.8058 0.4897 0.2298 0.2986 0.3826 中度耐盐Moderate salt tolerance
49 -0.4474 1.3137 -0.7449 0.2948 0.3656 0.3110 0.3183 中度耐盐Moderate salt tolerance
50 0.1973 -0.2087 0.3914 0.3649 0.1722 0.5432 0.3364 中度耐盐Moderate salt tolerance
51 0.6813 0.2845 0.1753 0.4176 0.2349 0.4990 0.3766 中度耐盐Moderate salt tolerance
52 0.3868 0.2984 -1.8436 0.3856 0.2366 0.0865 0.2940 盐敏感Salt sensitivity
53 0.3103 -0.0096 -0.0411 0.3772 0.1975 0.4548 0.3365 中度耐盐Moderate salt tolerance
54 -1.3353 1.3425 0.5085 0.1982 0.3693 0.5671 0.3074 中度耐盐Moderate salt tolerance
权重 Index weight(Wj) 0.5458 0.2951 0.1591

Fig. 1

Cluster of salt- tolerance in germination of 54 foxtail millet varieties"

Table 5

Changes of Relative Biomass under NaCl stress"

品种Variety 济谷16 Jigu 16 鲁谷1号Lugu 1 豫谷1号Yugu 1
相对生物量Relative biomass 0.888a 0.769b 0.926a

Fig. 2

Changes of SOD enzyme activity under NaCl stress ** indicates a significant relationship at 0.01 level; * indicates a significant relationship at 0.05 level. The same as below"

Fig. 3

Changes of POD enzyme activity under NaCl stress"

Fig. 4

Changes of CAT enzyme activity under NaCl stress"

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