Table I. MALDI-TOF MS identification of proteins preferentially accumulated in wild-type or in fl mutant cotton ovules.
Spot no.a | Protein name | NCBI Accession no. | pI/exp.b molecular mass (kDa) | pI/theo.c molecular mass (kDa) | Score/cov.d (%) | Matched/searchede | Relative protein content |
Ratio WT/FL | |
---|---|---|---|---|---|---|---|---|---|
WT-10 | FL-10 | ||||||||
1 | Profilin | ABO43717 | 5.33/13.01 | 5.38/14.41 | 72/56 | 10/60 | 0.397 ± 0.034 | 0.205 ± 0.026 | 1.94 |
2f | Major latex-like protein | FJ415202 | 5.44/14.83 | 5.46/17.16 | 86/60 | 10/86 | 0.236 ± 0.028 | 0.121 ± 0.014 | 1.95 |
3 | Annexin 1 | AAR13288 | 6.31/15.81 | 6.19/36.15 | 172/55 | 20/64 | 0.103 ± 0.017 | 0.017 ± 0.009 | 6.06 |
6 | Annexin | AAB67993 | 6.08/16.10 | 6.41/36.03 | 99/35 | 13/68 | 0.037 ± 0.005 | 0 | g |
18f | Annexin | FJ415173 | 6.52/28.49 | 6.74/35.98 | 123/38 | 14/60 | 0.065 ± 0.021 | 0.022 ± 0.008 | 2.95 |
5 | Fiber annexin | AAC33305 | 6.11/16.04 | 6.34/36.21 | 99/37 | 15/81 | 0.075 ± 0.015 | 0.038 ± 0.011 | 1.97 |
4f | Copper,zinc-superoxide dismutase | FJ415203 | 5.83/15.98 | 5.47/15.36 | 106/92 | 10/59 | 0.162 ± 0.021 | 0.084 ± 0.025 | 1.93 |
7f | Copper,zinc-superoxide dismutase | FJ415203 | 5.83/16.35 | 5.47/15.36 | 88/92 | 10/93 | |||
8f | Peroxiredoxin | FJ415174 | 5.35/17.41 | 5.58/17.30 | 140/74 | 10/44 | 0.127 ± 0.028 | 0.048 ± 0.008 | 2.65 |
9f | Dimethylmenaquinone methyltransferase | FJ415179 | 5.41/18.54 | 5.60/18.05 | 104/50 | 7/42 | 0.145 ± 0.023 | 0.080 ± 0.008 | 1.81 |
10 | Benzoquinone reductase | ABN12321 | 6.09/21.65 | 6.09/21.65 | 69/39 | 5/49 | 0.295 ± 0.013 | 0.053 ± 0.007 | 5.57 |
13 | Benzoquinone reductase | ABN12321 | 6.27/27.60 | 6.09/21.65 | 80/39 | 5/56 | |||
14 | Benzoquinone reductase | ABN12320 | 6.47/27.83 | 6.20/21.79 | 82/31 | 5/29 | 0.348 ± 0.039 | 0.034 ± 0.012 | 10.24 |
88f | Benzoquinone reductase | FJ415183 | 7.68/24.72 | 6.97/21.74 | 94/48 | 12/53 | 0.131 ± 0.019 | 0 | g |
11 | Ascorbate peroxidase | ABR18607 | 5.13/27.49 | 5.93/27.74 | 84/52 | 9/50 | |||
12 | Ascorbate peroxidase | ABR18607 | 5.43/27.51 | 5.93/27.74 | 79/44 | 7/100 | |||
15 | Ascorbate peroxidase | ABR18607 | 5.68/27.84 | 5.93/27.74 | 90/46 | 6/77 | |||
16 | Ascorbate peroxidase | ABR18607 | 5.29/27.87 | 5.93/27.74 | 78/47 | 9/59 | 0.239 ± 0.025 | 0.040 ± 0.017 | 5.98 |
19 | Ascorbate peroxidase | ABR18607 | 5.32/28.70 | 5.93/27.74 | 101/49 | 9/51 | |||
20 | Ascorbate peroxidase | ABR18607 | 5.64/28.75 | 5.93/27.74 | 151/64 | 15/41 | |||
21 | Ascorbate peroxidase | ABR18607 | 5.15/29.28 | 5.62/27.58 | 112/60 | 13/67 | |||
17f | Ascorbate peroxidase | FJ415185 | 4.96/27.89 | 5.62/27.58 | 66/38 | 5/60 | 0.207 ± 0.032 | 0.043 ± 0.015 | 4.81 |
38f | Stromal ascorbate peroxidase | FJ415186 | 6.24/36.78 | 8.89/41.05 | 85/35 | 13/75 | 0.102 ± 0.014 | 0.047 ± 0.015 | 2.17 |
40f | Stromal ascorbate peroxidase | FJ415186 | 6.10/37.04 | 8.89/41.05 | 73/28 | 9/100 | |||
22f | α-1,4-glucan phosphorylase | FJ415211 | 4.94/30.21 | 5.32/10.67 | 108/23 | 19/94 | 0.132 ± 0.010 | 0 | g |
23f | S-Formylglutathione hydrolase | FJ415188 | 6.59/30.65 | 6.82/32.18 | 77/47 | 10/89 | 0.120 ± 0.036 | 0.050 ± 0.002 | 2.40 |
24f | Triose-phosphate isomerase | FJ415177 | 6.53/31.54 | 6.00/27.47 | 142/82 | 15/110 | 0.253 ± 0.035 | 0.170 ± 0.019 | 1.49 |
25f | 20 S proteasome subunit α-1 | FJ415181 | 6.44/31.80 | 5.91/27.39 | 112/47 | 11/49 | 0.105 ± 0.009 | 0.035 ± 0.01 | 3.00 |
26f | Heat shock protein 70 | FJ415196 | 6.27/32.83 | 5.07/71.57 | 74/19 | 9/77 | 0.146 ± 0.010 | 0.036 ± 0.025 | 4.06 |
68f | Heat shock protein 70 | FJ415196 | 4.75/52.95 | 5.10/71.37 | 84/26 | 15/99 | |||
63f | Heat shock protein 70 | FJ415199 | 4.59/49.92 | 5.10/71.35 | 88/27 | 9/95 | 0.113 ± 0.007 | 0.043 ± 0.041 | 2.63 |
67f | Heat shock protein 70 | FJ415194 | 4.69/52.90 | 5.14/71.28 | 89/21 | 9/37 | 0.468 ± 0.027 | 0.134 ± 0.030 | 3.49 |
90f | Heat shock protein 70 | FJ415194 | 7.52/33.12 | 5.14/71.28 | 84/20 | 9/65 | |||
73f | Heat shock protein 70 | FJ415195 | 4.75/54.07 | 5.07/71.57 | 90/13 | 8/47 | 0.059 ± 0.007 | 0.010 ± 0.011 | 5.90 |
27f | Catalase | FJ415187 | 6.30/33.59 | 6.68/57.25 | 73/30 | 16/84 | 0.197 ± 0.032 | 0.091 ± 0.015 | 2.16 |
28f | Serine hydroxymethyltransferase | FJ415180 | 5.54/34.18 | 7.57/52.38 | 95/26 | 9/59 | 0.091 ± 0.008 | 0.014 ± 0.001 | 6.50 |
31f | Serine hydroxymethyltransferase | FJ415180 | 5.83/34.82 | 7.57/52.38 | 75/36 | 12/76 | |||
29f | Lactoylglutathione lyase | FJ415204 | 5.71/34.38 | 5.69/32.61 | 69/34 | 11/69 | 0.202 ± 0.016 | 0 | g |
30f | α-Soluble NSFh attachment protein | FJ415171 | 5.08/34.75 | 5.11/33.05 | 120/39 | 9/23 | 0.110 ± 0.033 | 0.034 ± 0.012 | 3.24 |
32f | UER1 | FJ415167 | 5.94/34.94 | 5.73/33.95 | 180/62 | 18/79 | 0.253 ± 0.030 | 0.253 ± 0.030 | 1.78 |
33f | UER1 | FJ415167 | 6.22/34.97 | 5.73/33.95 | 215/64 | 15/31 | |||
34f | Fructokinase | FJ415169 | 5.05/36.08 | 5.28/35.20 | 206/58 | 16/41 | 0.080 ± 0.002 | 0.045 ± 0.009 | 1.78 |
35 | Enolase | ABW21688 | 5.21/36.28 | 5.49/47.98 | 85/29 | 7/78 | 0.049 ± 0.003 | 0 | g |
36 | Actin | AAP73454 | 5.43/36.64 | 5.23/41.90 | 90/46 | 12/63 | 0.178 ± 0.027 | 0.027 ± 0.028 | 6.59 |
52 | Actin | AAP73452 | 5.62/42.35 | 5.44/41.94 | 69/29 | 7/86 | 0.264 ± 0.017 | 0.018 ± 0.020 | 14.67 |
53 | Actin | AAP73457 | 5.56/43.29 | 5.31/41.91 | 149/60 | 28/142 | 0.244 ± 0.03 | 0.039 ± 0.036 | 6.26 |
71 | Actin | AAP73460 | 5.45/53.58 | 5.37/41.94 | 163/57 | 22/91 | 0.189 ± 0.023 | 0.120 ± 0.032 | 1.58 |
37f | Granule-bound starch synthase | FJ415189 | 4.97/36.64 | 8.79/63.84 | 80/20 | 8/55 | 0.087 ± 0.012 | 0 | g |
39f | Granule-bound starch synthase | FJ415205 | 4.91/36.78 | 8.59/67.73 | 135/35 | 13/89 | 0.090 ± 0.016 | 0.019 ± 0.011 | 4.76 |
41f | Glutamine synthase | FJ415178 | 5.64/37.12 | 5.77/39.36 | 140/43 | 10/50 | 0.078 ± 0.006 | 0 | g |
42f | Malate dehydrogenase | FJ415192 | 6.14/38.50 | 6.10/36.45 | 93/27 | 10/77 | 0.337 ± 0.004 | 0.222 ± 0.028 | 1.52 |
56f | Malate dehydrogenase | FJ415192 | 6.67/46.15 | 6.10/35.87 | 88/43 | 8/100 | |||
43 | Phenylcoumaran benzylic ether reductase-like protein | ABN12322 | 5.58/38.89 | 5.76/33.89 | 92/35 | 11/48 | 0.227 ± 0.021 | 0.102 ± 0.022 | 2.23 |
51 | Phenylcoumaran benzylic ether reductase-like protein | ABN12322 | 5.87/41.46 | 5.76/33.89 | 129/58 | 18/97 | |||
45 | β-Tubulin 19 | ABY86665 | 5.75/39.76 | 4.76/50.65 | 173/58 | 26/70 | 0.073 ± 0.034 | 0 | g |
46 | α-Tubulin 4 | AAN33000 | 5.57/39.90 | 5.36/34.41 | 207/64 | 19/40 | 0.145 ± 0.007 | 0.059 ± 0.018 | 2.46 |
49 | α-Tubulin 4 | AAN33000 | 5.43/40.90 | 5.36/34.41 | 107/44 | 12/65 | |||
48 | α-Tubulin | ABO47738 | 5.72/40.45 | 4.97/50.29 | 159/43 | 19/38 | 0.155 ± 0.029 | 0 | g |
50f | Glyceraldehyde-3-phosphate dehydrogenase C subunit | FJ415206 | 6.61/41.24 | 7.70/36.65 | 73/33 | 7/87 | 0.234 ± 0.030 | 0.115 ± 0.052 | 2.04 |
54f | 2-Nitropropane dioxygenase | FJ415176 | 5.43/43.73 | 5.32/36.17 | 211/68 | 18/62 | 0.330 ± 0.009 | 0.134 ± 0.033 | 2.46 |
55f | Quinone oxidoreductase | FJ415175 | 5.21/45.90 | 5.28/34.39 | 149/67 | 16/106 | 0.088 ± 0.014 | 0 | g |
57 | Gibberellin 20-oxidase 1 | ABA01482 | 5.23/48.01 | 5.35/41.72 | 215/64 | 24/84 | 0.030 ± 0.007 | 0 | g |
58 | Flavanone 3-hydroxylase | ABM64799 | 5.33/48.10 | 5.43/41.75 | 171/66 | 26/130 | 0.485 ± 0.007 | 0.226 ± 0.023 | 2.14 |
59f | Mannitol dehydrogenase | FJ415191 | 5.93/49.11 | 5.85/39.57 | 100/61 | 16/90 | 0.252 ± 0.032 | 0.144 ± 0.026 | 1.75 |
60f | Adenosine kinase | FJ415170 | 5.46/49.66 | 5.47/37.81 | 200/59 | 15/38 | 0.059 ± 0.018 | 0.145 ± 0.028 | 1.40 |
61f | Adenosine kinase | FJ415170 | 5.31/49.66 | 5.47/37.81 | 128/55 | 18/64 | g | ||
62f | Phosphoglycerate dehydrogenase | FJ415190 | 5.20/49.84 | 7.14/64.06 | 81/23 | 7/54 | 0.111 ± 0.009 | 0 | g |
65f | Phosphoglycerate dehydrogenase | FJ415190 | 5.13/50.35 | 7.14/64.06 | 77/27 | 8/100 | |||
64f | Pyruvate dehydrogenase α subunit | FJ415197 | 6.74/50.17 | 7.16/43.69 | 79/30 | 9/100 | 0.222 ± 0.013 | 0.157 ± 0.008 | 1.41 |
66 | Anthocyanidin reductase | ABM64802 | 5.53/51.74 | 5.54/36.54 | 138/49 | 16/86 | 0.358 ± 0.003 | 0 | g |
69f | Luminal binding protein | FJ415200 | 4.57/53.09 | 5.13/73.57 | 81/31 | 17/95 | 0.062 ± 0.016 | 0.015 ± 0.012 | 4.13 |
70f | Luminal binding protein | FJ415200 | 4.50/53.55 | 5.13/73.57 | 76/23 | 9/100 | |||
72f | Luminal binding protein | FJ415200 | 4.55/53.94 | 5.13/73.57 | 104/25 | 10/67 | |||
74f | Phosphoglycerate kinase | FJ415172 | 6.14/56.40 | 5.97/42.29 | 140/44 | 14/59 | 0.063 ± 0.008 | 0.035 ± 0.003 | 1.80 |
75 | Chloroplast biotin carboxylase | ABP98813 | 6.30/57.92 | 7.57/59.17 | 94/34 | 19/85 | 0.083 ± 0.011 | 0.049 ± 0.010 | 1.69 |
77 | Chloroplast biotin carboxylase | ABP98813 | 6.31/59.64 | 7.57/59.17 | 139/61 | 26/101 | |||
76f | Dihydrolipoamide dehydrogenase | FJ415193 | 6.62/59.37 | 6.93/54.13 | 70/25 | 8/100 | 0.118 ± 0.026 | 0 | g |
78f | UGP2 | FJ415165 | 5.54/60.41 | 5.62/51.45 | 155/49 | 18/100 | 0.205 ± 0.015 | 0.145 ± 0.023 | 1.42 |
79f | UGP1 | FJ415164 | 6.07/61.70 | 5.81/51.74 | 132/52 | 17/61 | 0.178 ± 0.016 | 0.092 ± 0.014 | 1.93 |
80f | UGP1 | FJ415164 | 6.10/61.70 | 5.81/51.74 | 216/61 | 22/63 | |||
81f | UGD1 | FJ415166 | 6.11/62.84 | 5.84/53.64 | 183/59 | 23/109 | 0.124 ± 0.016 | 0.057 ± 0.013 | 2.18 |
83f | UGD1 | FJ415166 | 6.30/64.12 | 5.84/53.64 | 188/53 | 18/49 | |||
82f | myo-Inositol-1-phosphate synthase | FJ415168 | 5.69/63.53 | 5.46/56.54 | 207/54 | 24/106 | 0.090 ± 0.006 | 0.046 ± 0.010 | 1.96 |
84 | Acyltransferase-like protein | AAL67994 | 5.67/64.47 | 5.67/48.29 | 107/40 | 22/108 | 0.100 ± 0.013 | 0.019 ± 0.008 | 5.26 |
85 | Acyltransferase-like protein | AAL67994 | 5.56/64.82 | 5.67/48.29 | 72/38 | 15/110 | |||
86f | Pyruvate decarboxylase | FJ415201 | 6.57/69.52 | 6.13/60.77 | 99/30 | 11/100 | 0.128 ± 0.033 | 0.025 ± 0.014 | 5.00 |
87f | Glycine-rich RNA-binding protein | FJ415184 | 7.82/15.01 | 7.82/17.08 | 111/62 | 13/72 | 0.116 ± 0.030 | 0 | g |
89 | Manganese-superoxide dismutase | AAC78469 | 7.49/25.40 | 8.54/22.14 | 158/75 | 16/100 | 0.089 ± 0.015 | 0.041 ± 0.008 | 2.17 |
91f | Glyceraldehyde-3-phosphate dehydrogenase | FJ415182 | 7.74/43.72 | 7.06/37.04 | 92/58 | 15/100 | 0.167 ± 0.023 | 0.048 ± 0.015 | 3.48 |
92f | Isocitrate dehydrogenase | FJ415198 | 7.32/53.15 | 6.29/46.41 | 188/63 | 28/100 | 0.381 ± 0.030 | 0.256 ± 0.029 | 1.49 |
93f | Isocitrate dehydrogenase | FJ415198 | 7.07/53.29 | 6.29/46.41 | 140/57 | 23/100 | |||
94f | Eukaryotic translation initiation factor 5A | GU295062 | 5.55/18.88 | 5.61/17.63 | 96/52 | 9/61 | 0.156 ± 0.026 | 0.293 ± 0.024 | 0.53 |
95f | Chalcone isomerase | GU295063 | 4.84/29.85 | 4.85/23.42 | 95/67 | 10/66 | 0.104 ± 0.021 | 0.177 ± 0.017 | 0.59 |
96f | Triose-phosphate isomerase | GU295064 | 5.52/30.18 | 6.66/33.50 | 101/53 | 16/138 | 0.051 ± 0.009 | 0.122 ± 0.016 | 0.42 |
97f | Thiazole biosynthetic enzyme | GU295068 | 5.03/36.81 | 5.64/38.24 | 170/63 | 17/49 | 0.048 ± 0.008 | 0.155 ± 0.024 | 0.31 |
98f | Transaldolase | GU295065 | 4.91/45.56 | 5.78/43.06 | 129/33 | 10/18 | 0.036 ± 0.005 | 0.089 ± 0.012 | 0.40 |
100f | Transaldolase | GU295065 | 5.09/46.62 | 5.78/43.06 | 123/29 | 12/32 | |||
99f | U2 small nuclear ribonucleoprotein A | GU295066 | 5.06/45.90 | 4.97/32.19 | 204/68 | 18/50 | 0.062 ± 0.003 | 0.237 ± 0.033 | 0.26 |
101 | Chalcone synthase | ABS52573 | 6.11/54.17 | 6.12/42.98 | 87/35 | 11/45 | 0.039 ± 0.010 | 0.104 ± 0.026 | 0.38 |
102 | Protein-disulfide isomerase | ABO41843 | 5.01/65.30 | 5.07/55.89 | 203/59 | 24/77 | 0.051 ± 0.010 | 0.102 ± 0.003 | 0.50 |
103f | RNA helicase-like protein | GU295067 | 5.91/69.24 | 5.65/56.15 | 201/43 | 17/32 | 0.062 ± 0.012 | 0.121 ± 0.020 | 0.51 |
104 | Betaine-aldehyde dehydrogenase | AAR23816 | 5.44/69.43 | 5.60/55.37 | 100/42 | 19/103 | 0.055 ± 0.011 | 0.114 ± 0.012 | 0.48 |
a Protein spots are arranged from lowest to highest molecular mass with spots encoded by the same cotton cDNA quantified as one protein and spots presumably encoded by the same gene family located next to each other. Spots 1–93 were preferentially accumulated in the wild type, and spots 94–104 were preferentially accumulated in the mutant.
b Experimental.
c Theoretical.
d Coverage.
e Number of matched/searched polypeptides.
f Sixty-eight polypeptides encoded by 50 putative full length and one partial (spot 22, FJ415211) cotton cDNAs obtained for the first time in the current work.
g Protein spots only observed in WT-10.
h N-Ethylmaleimide-sensitive factor.