Table 2.
Fatty Acid Hydroxide Analysis of Germinating Seedlings
| Genotype | 13-HODE cis/trans | 13-HODE trans/trans | 13-HOTE | 9-HODE cis/trans | 9-HODE trans/trans | ΣLOH |
|---|---|---|---|---|---|---|
| 0 d | ||||||
| Col | NDa | ND | ND | ND | ND | ND |
| vte1-1 | ND | ND | ND | ND | ND | ND |
| vte2-1 | 0.4 ± 0.3 | 0.3 ± 0.3 | ND | 0 ± 0.3 | 0 ± 0.2 | 0.3 ± 0.5 |
| Ws | ND | ND | ND | ND | ND | ND |
| vte2-2 | 1.4 ± 0.6 | 1.8 ± 0.9 | ND | 1.0 ± 0.5 | 0.8 ± 0.5 | 2.6 ± 1.3 |
| 3 d | ||||||
| Col | 0.4 ± 0.2 | 0.6 ± 0.3 | ND | 0.3 ± 0.2 | ND | 0.4 ± 0.1 |
| vte1-1 | 0.6 ± 0.1 | 0.3 ± 0.2 | 0 ± 0.1 | 0.3 ± 0.1 | 0 ± 0.1 | 0.4 ± 0.2 |
| vte2-1 | 27.7 ± 2.7 (53:47) | 34.5 ± 5.4 | 2.2 ± 0.8 (56:44) | 23.0 ± 0.1 (50:50) | 20.1 ± 2.4 | 48.3 ± 6.0 |
| Ws | 0.5 ± 0.2 | ND | ND | ND | ND | 0.1 ± 0.1 |
| vte2-2 | 25.6 ± 1.3 (52:48) | 30.0 ± 2.5 | 1.3 ± 0.5 (60:40) | 20.5 ± 1.6 (51:49) | 16.7 ± 1.1 | 46.6 ± 3.8 |
| 6 d | ||||||
| Col | 1.2 ± 0.2 (51:49) | 1.0 ± 0.7 | 0.1 ± 0.1 | 0.6 ± 0.4 (52:48) | 0.6 ± 0.4 | 0.8 ± 0.4 |
| vte1-1 | 0.8 ± 0.1 (54:46) | 0.9 ± 0.2 | ND | 0.6 ± 0.1 (50:50) | 0.3 ± 0.2 | 0.7 ± 0.1 |
| vte2-1 | 31.3 ± 6.3 (52:48) | 30.0 ± 13.9 | 0.5 ± 0.9 (55:45) | 28.0 ± 4.6 (51:49) | 24.9 ± 9.2 | 42.6 ± 13.2 |
| Ws | 0.3 ± 0.1 (52:48) | 0.4 ± 0.2 | 0.0 ± 0.1 | 0.1 ± 0.2 (47:53) | 0.3 ± 0.2 | 0.2 ± 0.1 |
| vte2-2 | 30.8 ± 3.9 (52:48) | 27.1 ± 6.0 | 0.3 ± 0.3 (56:44) | 26.1 ± 3.1 (51:49) | 19.8 ± 1.3 | 42.1 ± 6.2 |
The five prevalent hydroxy fatty acid species were quantified using normal phase HPLC. Each compound is expressed as pmol/nmol of the corresponding PUFA (linoleic or linolenic acid) from which it was derived in the sample. The enantiomeric composition of 9-HODE cis/trans, 13-HOTE, and 13-HODE cis/trans were determined by chiral phase HPLC. The ratio of R to S is given in parentheses for samples that contained sufficient quantities for chiral analysis. Chiral analysis indicated that both 9- and 13-HODE trans/trans peaks contained two enantiomers in relatively equal proportions, though R and S could not be assigned because of the lack of commercially available standards. ΣLOH is sum of all LOHs expressed as pmol/nmol total PUFAs (linoleic and linolenic acid). The zero day time point is seed that have been imbibed for 5 d at 4°C. The data are the means ± sd (n = 4). The LOH values for both vte2-1 and vte2-2 were highly significant (Student's t test, P < 0.01) at all points relative to the wild type.
ND, not detected.