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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (9): 1431-1439.doi: 10.3724/SP.J.1006.2019.81088

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Effects of salt stress on physiological indexes and differential proteomics of oat leaf

CHEN Xiao-Jing,LIU Jing-Hui(),YANG Yan-Ming,ZHAO Zhou,XU Zhong-Shan,HAI Xia,HAN Yu-Ting   

  1. Inner Mongolia Agricultural University/Cereal Engineering Technology Research Center, Inner Mongolia Autonomous Region/ National Agricultural Research Outstanding Talents and Innovation Team, Huhhot 010019, Inner Mongolia, China
  • Received:2018-12-13 Accepted:2019-05-12 Online:2019-09-12 Published:2019-06-06
  • Contact: Jing-Hui LIU E-mail:cauljh@163.com
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31560357)

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Abstract:

MDA content, SOD and POD activities, free proline content of oat leaves were determined under salt stress (molar NaCl: Na2SO4=1:1), and the differentially expressed proteins in leaves were analyzed by using Label-Free technique. The results showed that the activities of MDA, SOD and POD in oat leaves decreased by 16.7%, 23.4%, and 21.2% respectively, and free proline content increased by 1.12% compared with the control. There were 76 differential proteins with P-value ≤ 0.05 and ratio > 2 (51 proteins up-regulated and 25 proteins down-regulated). The GO annotation indicated that 27 differential proteins were significantly enriched in 16 metabolic pathways, of which the oxidation-reduction process was 33.9%. The biological processes of level 3 statistical enrichment were oxygen binding and oxidoreductase activity. Twenty-two differential proteins were obtained using KEGG annotation, which was significantly enriched in 10 biochemical metabolic pathways, mainly including four processes: protein processing in endoplasmic reticulum, longevity regulating pathway-multiple species, antigen processing and presentation, estrogen signaling pathway. The STRING protein interaction network showed that 10 of the 21 differential proteins involved in post-translational modification, protein turnover, and molecular chaperone function, and HSP70 and HSP90 interacted with the most core proteins of the whole network. It is speculated that the up-regulation of core protein is one of the reasons for the salt tolerance of oats.

Key words: salt stress, oat, Label-Free, proteomic

Fig. 1

Effect of salt stress on antioxidant enzyme activity Bars superscripted by different letters are significantly different between treatments (P < 0.05)."

Fig. 2

Differential protein volcano map The abscissa indicates the difference fold (log2 value) of the differential protein, the ordinate indicates P-value (-lg value), black indicates the protein with insignificant difference, red indicates the up-regulated protein, and green indicates the down-regulated protein."

Fig. 3

Differential protein clustering heat map Vertical direction shows the clustering of samples, and horizontal direction shows the clustering of proteins. Pattern clustering of protein content between samples can be seen from longitudinal clustering."

Table 1

BYC and BYS differential proteins"

功能类
Functional class		 蛋白名称
Protein ID		 功能描述
Functional description		 COG 编号
COG gene ID		 可信度
Identity		 上调/下调
Up(↑)/Down(↓)
能量生产和转换 Energy production and conversion (9)
C A0A193CI13 FoF1-type ATP synthase, beta subunit YP_722959 0.91 ↑
C A9LIN4 Malic enzyme YP_007218386 0.53 ↑
C
 I1GZ41
 Acyl-CoA reductase or other NAD-dependent aldehyde dehydrogenase YP_722268
 0.61
 ↑
C A0A1V0EL65 Hemoglobin-like flavoprotein YP_007103351 0.30 ↓
C A0A1D5S6L5 Malic enzyme YP_005607828 0.45 ↑
C A0A1D6CUL3 NADH dehydrogenase, FAD-containing subunit YP_003322333 0.33 ↑
C I1I051 Coenzyme F420-reducing hydrogenase, beta subunit YP_007131148 0.55 ↓
C A0A1D6AN16 Ferredoxin-NADP reductase YP_007118319 0.52 ↓
C W4ZQA0 Hemoglobin-like flavoprotein YP_003692354 0.33 ↓
翻译后修饰、蛋白质周转和分子伴侣 Posttranslational modification, protein turnover, and chaperones (15)
O A0A165FYR3 Glutathione S-transferase YP_007096787 0.31 ↑
O Q3I0N4 Molecular chaperone IbpA, HSP20 family YP_005887446 0.38 ↑
O A0A1C6ZYA4 Molecular chaperone, HSP90 family YP_634186 0.45 ↑
O I1H6R7 Glutathione S-transferase YP_007063563 0.28 ↑
O F4Y589 Molecular chaperone, HSP90 family YP_634186 0.47 ↑
O A0A1D5Y3B7 ATP-dependent Clp protease ATP-binding subunit ClpA YP_007121403 0.69 ↑
O I1IF07 Molecular chaperone IbpA, HSP20 family YP_001357091 0.31 ↑
O M8BCN0 Molecular chaperone DnaK (HSP70) YP_005440675 0.49 ↑
O I1GZ93 Molecular chaperone IbpA, HSP20 family YP_522114 0.38 ↑
O M0Y631 Chaperonin GroEL (HSP60 family) YP_006371119 0.62 ↓
O F2E3N4 FKBP-type peptidyl-prolyl cis-trans isomerase YP_007057296 0.66 ↑
O M8AN59 ATP-dependent Zn proteases YP_723906 0.37 ↑
O F2DYT5 Molecular chaperone DnaK (HSP70) YP_005440675 0.49 ↑
O A0A1D5SA32 ATP-dependent Clp protease ATP-binding subunit ClpA YP_007132111 0.52 ↑
O W5EGU4 Molecular chaperone DnaK (HSP70) YP_423803 0.71 ↑
氨基酸转运和代谢; 辅酶转运和代谢; 一般功能预测; 次生代谢产物的生物合成、转运和分解代谢
Amino acid transport and metabolism; Coenzyme transport and metabolism; General function prediction only; Secondary metabolites biosynthesis, transport, and catabolism (15)
E A0A1D5XXT6 Monoamine oxidase YP_005086783 0.30 ↓
E A0A1D6CAG8 Monoamine oxidase YP_005086783 0.29 ↓
E A0A1D5YV26 3-dehydroquinate dehydratase NP_867287 0.36 ↑
E I1GLP9 5,10-methylenetetrahydrofolate reductase YP_112676 0.40 ↑
E A0A1D6BBP7 Aminopeptidase N YP_006773912 0.36 ↑
E I1HGT7 Threonine synthase YP_002463167 0.62 ↑
EH I1GU32
 3'-phosphoadenosine 5'-phosphosulfate sulfotransferase (PAPS reductase)/FAD synthetase or related enzyme YP_007109372
 0.63
 ↓
H I1HWV1 Glutamine amidotransferase PdxT (pyridoxal biosynthesis) YP_005441479 0.52 ↑
HR F2CYS4 Hydroxymethylpyrimidine pyrophosphatase and other HAD family phosphatases YP_007159391
 0.34
 ↑
R A0A1D6APK4 Zn-dependent alcohol dehydrogenase YP_001983794 0.67 ↓
功能类
Functional class		 蛋白名称
Protein ID		 功能描述
Functional description		 COG 编号
COG gene ID		 可信度
Identity		 上调/下调
Up(↑)/Down(↓)
R T1MRH6 Predicted oxidoreductase (related to aryl-alcohol dehydrogenase) YP_007098133
 0.45
 ↑
R W4ZPA5 Uncharacterized metalloenzyme YdcJ, glyoxalase superfamily YP_628605
 0.26
 ↑
QR A0A1D5VID8 NADPH-dependent curcumin reductase CurA YP_003953479 0.52 ↑
Q I1ICZ4 Carotenoid cleavage dioxygenase or a related enzyme YP_007064275 0.35 ↓
Q A0A1D5UEP5 Cu2+-containing amine oxidase YP_005086886 0.34 ↑
碳水化合物的运输和代谢 Carbohydrate transport and metabolism (7)
G
 Q38786
 Beta-glucosidase/6-phospho-beta-glucosidase/beta-galactosidase YP_004449345
 0.40
 ↓
G Q8S311 Sucrose-6-phosphate hydrolase SacC, GH32 family YP_002315570 0.29 ↑
G M0WF67 6-phosphogluconate dehydrogenase YP_007142557 0.47 ↓
G I1IJ14 Ribulose bisphosphate carboxylase small subunit YP_007111326 0.63 ↑
G I1I5R4 Predicted arabinose efflux permease, MFS family YP_003405887 0.40 ↑
G A0A1D6RIU0 1,4-alpha-glucan branching enzyme YP_677957 0.47 ↑
G Q7X9A2 Ribulose bisphosphate carboxylase small subunit YP_007111326 0.63 ↑
翻译、核糖体结构和生物发生 Translation, ribosomal structure, and biogenesis (5)
J M0WGV4 Ribosome-associated translation inhibitor RaiA YP_007127538 0.38 ↓
J I1GM81 Ribosomal protein L4 NP_275148 0.41 ↓
J F2DBP2 Ribosomal protein L6P/L9E YP_007126712 0.54 ↑
J I1IU29 Ribosomal protein L3 YP_004004412 0.39 ↓
J M8CXZ0 RNA recognition motif (RRM) domain YP_844222 0.44 ↓
无机离子转运和代谢 Inorganic ion transport and metabolism (2)
P I1I9J4 Cu/Zn superoxide dismutase YP_003290884 0.49 ↓
P R7WAY7 Carbonic anhydrase YP_004682419 0.38 ↓
脂质运输和新陈代谢 Lipid transport and metabolism (3)
I M7ZQT4 Isopentenyldiphosphate isomerase YP_006428266 0.39 ↑
I I1IT00
 NADPH-dependent2,4-dienoyl-CoA reductase, sulfur reductase, or a related oxidoreductase YP_824941
 0.32
 ↑
I O65195 Myo-inositol-1-phosphate synthase YP_005007067 0.35 ↓
信号转导机制 Signal transduction mechanisms (2)
T I1J3C6 Phosphohistidine swiveling domain of PEP-utilizing enzymes YP_002508078
 0.47
 ↓
T I1HYN7 Predicted NTPase, NACHT family domain YP_003890644 0.30 ↑
细胞壁/膜/包膜生物发生 Cell wall/membrane/envelope biogenesis (2)
M W5BNP0 Glycosyltransferase involved in cell wall bisynthesis YP_004596549 0.26 ↑
M I1I6H2 Nucleoside-diphosphate-sugar epimerase YP_007093680 0.72 ↑
防御机制 Defense mechanisms (1)
V I1GTZ4
 Enamine deaminase RidA, house cleaning of reactive enamine intermediates, YjgF/YER057c/UK114 family YP_003319835
 0.55
 ↑
功能未知 Function unknown (1)
S I1IC12 Uncharacterized protein YjbI, contains pentapeptide repeats YP_007125802 0.50 ↑

Fig. 4

Differential protein GO enrichment results The enrichment results in the three categories are shown in the figure, with up to 20 of each."

Fig. 5

Bubble diagram of differential protein KEGG enrichment results The abscissa is the ratio of the number of differential proteins in the corresponding term to the number of total proteins identified. The larger the value, the higher the degree of differential protein enrichment in this term. The color of the point represents the P-value of the hypergeometric test. The smaller the value, the greater the reliability and statistical significance of the test. The size of the dots represents the number of differential proteins in the corresponding term, the larger the size the more differential protein number in the term."

Fig. 6

Protein interaction network diagram The red node represents the up-regulated protein, the blue node represents the down-regulated protein, and the larger the node, the more the number of proteins interacting with it, and the thickness of the line between the nodes represents the interaction strength."

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