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Table 2 DAP spots and their identification by MALDI-TOF/TOF-MS in leaves from C. sinensis seedlings submitted to pH 2.5, pH 3 or pH 6 for 9 months

From: Low pH-responsive proteins revealed by a 2-DE based MS approach and related physiological responses in Citrus leaves

Spot No.a

Protein identity

Accession No.

Mr (kDa)/pI theor.

Mr (kDa)/pI exp.

Protein score

Peptide ions

NMPb

Ratioc

Coveredd sequence (%)

Charge

pH 2.5

pH 3

pH 6

Antioxidation and detoxification

 S6

L-ascorbate peroxidase 1, cytosolic

Cs8g17370.1

27.57/5.55

44.86/5.80

584

1 26

24

2.16 ± 0.09a

1.07 ± 0.12b

1.00 ± 0.03b

22

1

 S4

L-ascorbate peroxidase 3, peroxisomal

Cs3g19810.2

47.33/8.59

53.31/5.73

573

144

25

2.94 ± 0.80a

1.11 ± 0.21b

1.00 ± 0.15b

23

1

 S5

Probable aldo-keto reductase 1

Cs3g10670.1

38.42/5.50

62.64/5.68

295

90

15

2.41 ± 0.26a

0.84 ± 0.05b

1.00 ± 0.17b

14

1

 S8

Isoflavone reductase-like protein

Cs2g16220.1

34.25/6.40

56.42/6.23

155

56

17

2.03 ± 0.29a

0.80 ± 0.16b

1.00 ± 0.11b

15

1

 S1

Peroxidase 15

orange1.1 t02046.1

37.43/4.52

71.41/4.46

178

90

9

0.37 ± 0.07b

0.45 ± 0.06b

1.00 ± 0.04a

8

1

 S9

Glutathione S-transferase U19

Cs5g15190.1

25.56/7.56

42.77/6.64

148

71

14

0.30 ± 0.05b

0.83 ± 0.10a

1.00 ± 0.09a

13

1

 S10

Thioredoxin-2

Cs7g13660.1

21.44/8.44

21.81/4.87

178

62

7

0.47 ± 0.10b

0.87 ± 0.11a

1.00 ± 0.06a

6

1

Stress response

 S11

Heat shock protein 90–1

Cs5g03150.1

80.52/5.03

90.75/5.15

186

96

41

1.62 ± 0.19a

0.53 ± 0.09c

1.00 ± 0.02b

37

1

 S12

Putative uncharacterized protein Sb02g035950

Cs1g06050.1

27.76/5.56

52.74/6.00

265

87

11

1.29 ± 0.04a

0.51 ± 0.14b

1.00 ± 0.10a

10

1

 S13

Abscisic stress ripening-like protein

Cs3g21500.1

20.05/5.75

45.68/6.08

411

92

14

0.20 ± 0.05c

1.59 ± 0.11a

1.00 ± 0.06b

13

1

 S17

Thiamine thiazole synthase 1, chloroplastic

Cs4g11090.1

37.60/5.40

53.52/5.22

304

95

15

1.64 ± 0.12a

0.75 ± 0.02b

1.00 ± 0.18b

14

1

 S39

S-norcoclaurine synthase; Pathogenesis-related (PR)-10-related norcoclaurine synthase-like protein

Cs6g03210.1

17.29/4.89

28.52/5.12

485

90

17

4.45 ± 0.27a

2.23 ± 0.16b

1.00 ± 0.13c

15

1

Carbohydrate and energy metabolism

 S14

Chlorophyll a-b binding protein 215

Cs1g06360.1

28.93/5.13

41.21/4.90

71

56

6

2.22 ± 0.10a

0.46 ± 0.03c

1.00 ± 0.16b

5

1

 S20

Chlorophyll a-b binding protein 4, chloroplastic

Cs3g06180.1

29.52/6.84

40.18/5.42

167

88

11

0.13 ± 0.02c

0.71 ± 0.07b

1.00 ± 0.11a

10

1

 S16

Oxygen-evolving enhancer protein 1–1, chloroplastic

Cs1g23450.1

35.38/5.83

49.12/5.09

371

110

14

2.20 ± 0.59a

0.82 ± 0.11b

1.00 ± 0.02b

13

1

 S19

Oxygen-evolving enhancer protein 1–1, chloroplastic

Cs1g23450.1

35.38/5.83

48.88/5.44

523

135

19

2.67 ± 0.76a

0.99 ± 0.19b

1.00 ± 0.03b

17

1

 S18e

Ribulose bisphosphate carboxylase/oxygenase activase 1, chloroplastic

Cs7g31800.4

50.90/5.33

68.00/5.12

534

91

23

0.30 ± 0.06b

0.44 ± 0.06b

1.00 ± 0.09a

21

1

 S21

Ribulose bisphosphate carboxylase/oxygenase activase 1, chloroplastic

Cs7g31800.3

46.96/5.94

65.74/5.36

511

109

20

0.21 ± 0.02b

0.91 ± 0.02a

1.00 ± 0.08a

18

1

 S22

Rubisco subunit binding-protein β-2 subunit; Chaperonin 60 subunit β 1

Cs9g03300.1

64.78/5.85

81.19/5.36

798

107

39

0.33 ± 0.04b

1.10 ± 0.10a

1.00 ± 0.13a

35

1

 S35

Ferredoxin-NADP reductase, leaf isozyme, chloroplastic

Cs1g25510.1

40.48/8.68

55.49/6.40

258

80

22

0.45 ± 0.04c

0.65 ± 0.08b

1.00 ± 0.02a

20

1

 S37

Ferredoxin-NADP reductase, leaf isozyme, chloroplastic

Cs1g25510.4

40.48/8.68

55.29/6.70

240

82

21

0.45 ± 0.03b

0.91 ± 0.11a

1.00 ± 0.13a

19

1

 S27

Phosphoglycerate kinase 1, chloroplastic

orange1.1 t03280.1

49.45/8.20

67.19/5.91

567

95

25

0.25 ± 0.10b

0.87 ± 0.06a

1.00 ± 0.11a

23

1

 S29

Phosphoglycerate kinase 1, chloroplastic

orange1.1 t03280.1

49.45/8.20

80.46/5.95

346

92

13

0.06 ± 0.02b

0.95 ± 0.10a

1.00 ± 0.14a

12

1

 S23

2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase, chloroplastic

Cs3g01420.1

32.59/8.29

47.43/5.72

415

121

14

0.31 ± 0.04b

0.80 ± 0.08a

1.00 ± 0.15a

13

1

 S24

Succinyl-CoA ligase [ADP-forming] subunit beta, mitochondrial

Cs5g29390.1

45.26/5.98

63.82/5.62

286

78

27

0.52 ± 0.13b

1.10 ± 0.06a

1.00 ± 0.06a

25

1

 S28

Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial

Cs1g17930.1

59.37/8.43

77.39/5.80

390

94

23

0.30 ± 0.06c

0.77 ± 0.04b

1.00 ± 0.05a

21

1

 S32

Dihydrolipoyllysine-residue succinyltransferase component of 2-oxoglutarate dehydrogenase complex 1, mitochondrial

Cs2g21190.1

51.06/9.07

72.17/6.03

181

135

11

0.19 ± 0.04b

0.86 ± 0.04a

1.00 ± 0.07a

10

1

 S3

NADH dehydrogenase [ubiquinone] iron-sulfur protein 8-A, mitochondrial

Cs8g06410.1

26.11/5.68

42.59/5.12

238

48

20

2.58 ± 0.27a

1.10 ± 0.10b

1.00 ± 0.09b

18

1

 S26

ATP synthase gamma chain 1, chloroplastic

Cs2g03080.1

40.62/6.08

59.62/5.82

208

103

10

0.19 ± 0.04c

0.44 ± 0.06b

1.00 ± 0.06a

9

1

 S30

Probable ATP synthase 24 kDa subunit, mitochondrial

Cs1g04030.1

27.52/8.90

46.81/6.24

368

96

22

0.22 ± 0.06b

0.55 ± 0.11b

1.00 ± 0.14a

20

1

 S36

Probable ATP synthase 24 kDa subunit, mitochondrial

Cs1g04030.1

27.52/8.90

45.64/6.86

543

102

27

0.29 ± 0.07b

0.68 ± 0.08ab

1.00 ± 0.24a

25

1

 S47

DNA-damage-repair/toleration protein DRT102; ribose-5-phosphate isomerase B

Cs3g11320.1

33.52/5.25

53.47/5.39

393

85

19

0.44 ± 0.07b

1.02 ± 0.10a

1.00 ± 0.16a

17

1

Protein and amino acid metabolism

 S40

Peptidyl-prolyl cis-trans isomerase CYP37, chloroplastic

Cs1g06710.1

50.39/6.42

60.41/5.11

579

139

21

3.06 ± 0.91a

1.36 ± 0.22ab

1.00 ± 0.11b

19

1

 S43

T-complex protein 1 subunit beta

Cs3g26890.2

57.19/5.56

79.99/5.91

226

73

22

0.30 ± 0.13b

1.04 ± 0.05a

1.00 ± 0.19a

20

1

 S41

Proteasome subunit beta type-6-A like protein

Cs7g07630.1

25.57/5.21

40.62/5.73

254

86

12

0.23 ± 0.01c

0.68 ± 0.04b

1.00 ± 0.10a

11

1

 S44

26S proteasome non-ATPase regulatory subunit 11A

Cs4g04180.1

47.05/5.79

70.34/6.16

367

97

24

0.22 ± 0.028b

0.83 ± 0.091a

1.00 ± 0.06a

22

1

 S50

Unknown protein DS12 from 2D-PAGE of leaf, chloroplastic; ACT domain-containing protein, putative, expressed

orange1.1 t04488.1

30.69/5.59

51.57/4.75

333

105

15

0.35 ± 0.07b

0.89 ± 0.02a

1.00 ± 0.04a

14

1

 S51

Kynurenine formamidase

Cs8g05140.1

30.62/5.32

55.58/5.48

270

75

13

0.44 ± 0.01b

1.04 ± 0.03a

1.00 ± 0.08a

12

1

Cell wall and cytoskeleton

 S45

Tubulin alpha-1 chain

Cs9g03120.1

49.75/4.99

73.13/5.41

487

85

20

0.40 ± 0.05b

0.93 ± 0.12a

1.00 ± 0.13a

18

1

 S46

Caffeic acid 3-O-methyltransferase 1

orange1.1 t05216.1

29.98/5.73

63.37/6.19

194

66

11

0.53 ± 0.09b

1.18 ± 0.04a

1.00 ± 0.04a

10

1

Nucleic acid metabolism

 S48

Hypoxanthine-guanine phosphoribosyltransferase

Cs3g21990.1

20.48/5.54

38.60/5.61

143

94

8

0.41 ± 0.03b

1.07 ± 0.17a

1.00 ± 0.08a

7

1

Lipid metabolism

 S2

Plastid lipid-associated protein 2, chloroplastic; Chromoplast-specific carotenoid-associated protein, chromoplast

Cs2g02520.1

43.18/6.08

50.48/4.67

503

92

24

1.68 ± 0.09a

0.79 ± 0.18b

1.00 ± 0.02b

22

1

 S7

Epoxide hydrolase 4

Cs2g06360.1

36.94/5.90

59.20/6.19

352

73

21

2.51 ± 0.27a

0.90 ± 0.10b

1.00 ± 0.18b

19

1

 S31

Cinnamoyl-CoA reductase 1

Cs8g20610.1

35.48/5.57

55.34/6.06

220

77

14

0.31 ± 0.02b

0.66 ± 0.21ab

1.00 ± 0.12a

13

1

Cellular transport

 S15

Ferritin-1, chloroplastic

Cs6g09150.2

28.97/5.46

43.81/4.92

88

64

8

1.97 ± 0.34a

0.69 ± 0.09b

1.00 ± 0.11b

7

1

Signal transduction

 S38

14–3-3-like protein GF14 kappa; General regulatory factor 8

Cs1g20220.1

27.69/4.87

45.64/4.90

339

106

20

4.71 ± 0.32a

1.20 ± 0.15b

1.00 ± 0.06b

18

1

Others

 S52

Putative uncharacterized protein Sb02g003450

Cs5g09380.2

27.69/7.84

45.27/5.58

193

60

14

0.38 ± 0.11b

1.00 ± 0.09a

1.00 ± 0.12a

13

1

 S42

Putative uncharacterized protein Sb09g010000

Cs8g19010.1

55.68/7.66

79.38/5.66

248

102

19

0.19 ± 0.06b

0.73 ± 0.13a

1.00 ± 0.22a

17

1

 S34

Flavoprotein WrbA

Cs4g11860.1

22.29/5.75

40.45/6.51

256

136

10

0.44 ± 0.05b

0.67 ± 0.07ab

1.00 ± 0.14a

9

1

 S49

NAD(P)H-dependent 6′-deoxychalcone synthase

orange1.1 t00001.2

67.34/9.06

57.67/6.61

247

100

20

0.46 ± 0.07b

0.67 ± 0.11b

1.00 ± 0.07a

18

1

Unidentified protein spots

 S33

Probable phosphoglucomutase, cytoplasmic 1

orange1.1 t05474.1

16.20/5.32

84.77/6.25

100

91

4

0.40 ± 0.05b

0.77 ± 0.08ab

1.00 ± 0.17a

4

1

 S25

Glucose-6-phosphate 1-dehydrogenase 1, chloroplastic

Cs7g11110.1

66.95/7.68

25.37/5.76

65

23

21

0.46 ± 0.04b

1.34 ± 0.08a

1.00 ± 0.23b

19

1

 S53

Endo-1,3;1,4-beta-D-glucanase

Cs9g05910.3

28.00/7.07

43.84/5.09

43

/

12

3.32 ± 0.29a

1.02 ± 0.07b

1.00 ± 0.09b

11

1

 S54

E3 ubiquitin-protein ligase MARCH9

Cs9g05100.7

32.53/8.56

16.93/6.1

44

/

12

4.26 ± 0.19a

0.45 ± 0.02b

1.00 ± 0.18b

11

1

 S55

Cytochrome b6-f complex iron-sulfur subunit, chloroplastic

Cs2g22650.3

22.32/8.76

30.95/8.76

52

36

6

0.33 ± 0.04c

0.75 ± 0.05b

1.00 ± 0.09a

5

1

  1. aSpot number corresponds to the 2-DE imagines in Fig. 5
  2. bNMP means the number of matched peptides
  3. cRatio means the ratio of pH 2.5 and pH 3.0 to pH 6.0; Different letters within a row indicate significant differences at P < 0.05
  4. dCovered sequence (%) means the ratio of the number of amino of the matched peptides to the number of amino acids of the full-length protein
  5. eLow pH-responsive proteins shared by the two Citrus species were highlighted in bold