Table 2.
Identification of two-dimensional separated protein spots from rice coleoptile tips (top 7–11 mm) that differed in abundance between anoxic and aerated treatment
| Spot no. |
Gene indices |
Description |
MP |
Cov |
Score |
Match (MM/pI) |
Gel (MM/pI) mol. wt/PI |
Fold change (72 h anoxia/aerated) |
|---|---|---|---|---|---|---|---|---|
| 1 | 20279443 | Elicitor-inducible protein | 9 | 38% | 266 | 17 509/8·42 | 14 kDA/6·5 | 3, 2·4, 5·2, 2·4 |
| 2 | 13129480 | Unknown protein | 16 | 37% | 248 | 17 552/7·66 | 15 kDA/6·5 | 4·6, 3·8, >10, >10 |
| 3 | 585551 | Nucleoside diphosphate kinase (NDPK) | 21 | 57% | 474 | 16 851/6·30 | 16 kDA/6·6 | 1·3, 1·8, 3·4, >10 |
| 4 | 7619799 or 14090367 | Bowman–Birk trypsin inhibitor | 5 | 10% | 108 | 27 766/5·36 | 23 kDA/6·2 | 0·3, 0·5, <0·1 |
| 5 | 14090367 or 7619799 | Bowman–Birk trypsin inhibitor | 6 | 23% | 82 | 27 716/5·36 | 24 kDA/6·0 | 0·2, 0·4, <0·1 |
| 6 | 2331131 or 2293480 | Glycine-rich protein | 11 | 19% | 148 | 15 862/7·79 | 16kDA/5·4 | 0·1, 0·26, <0·1, <0·1 |
| 7 | 7433348 | Mn-superoxide dismutase (Mn-SOD) | 14 | 72% | 557 | 24 982/6·50 | 24 kDA/6·0 | 2·1, 1·5, 1·4 |
| 8 | 3550549 | Pyruvate orthophosphate dikinase (PPDK) | 4 | 5% | 87 | 96 504/5·42 | 96 kDA/5·2 | >10, 3·2, 3·8, 5·5 |
| 9 | 6979319 | Alcohol dehydrogenase 2 (ADH2) | 3 | 13% | 137 | 41 180/6·04 | 42 kDA/6·3 | – |
| 10 | 113360 | Alcohol dehydrogenase 1 (ADH1) | 5 | 14% | 140 | 40 826/6·32 | 42 kDA/6·5 | – |
| 11 | 7436606 | Fructose 1,6-bisphosphate aldolase | 4 | 16% | 82 | 38 799/8·35 | 33 kDA/6·4 | – |
| 12 | 2293480 or 1221131 | Glycine-rich protein | 4 | 24% | 121 | 16 534/8·97 | 16 kDA/5·4 | 0·3, 0·8, 0·9, 0·55 |
| 13 | 29124123 | Actin-depolymerizing factor | 14 | 65% | 242 | 15 936/5·72 | 16 kDA/5·2 | 1·2, 0·8, 1·5, 0·8 |
Tips were either dissected from intact seedlings which had been exposed to anoxia, or excised and healed before anoxia was imposed with 50 mm glucose. Numbered spots were derived from gels in Figs 1 and 3. MS/MS spectra derived from trypsinated peptides of proteins were matched at Mascot against a translated NCBI database (P < 0·05). MP = number of peptides matched; MM = molecular mass from gel or predicted from matched gene product sequence; pI = isoelectric point from gel or predicted from matched gene product sequence; Cov = percentage coverage of the matched sequence with the matched peptides; Score = Mascot match score, protein score >50 (P < 0·05). Fold change is the ratio of spot intensity following 72 h anoxia treatment in 50 mm glucose with the spot intensity in aerated treatment (n = 3 or 4 from Figs 1 and 3 and other independent experiments (gels not shown). Spots 9–11 could only be detected as [35S]methionine-labelled proteins from Fig. 3.