Wang et al. 10.1073/pnas.0603571103. |
Supporting Figure 6
Supporting Table 1
Supporting Figure 7
Supporting Figure 8
Fig. 6. Characterization of the AtFAAH insertional mutants and overexpressors. (A) The locations of the T-DNA insertions in SALK_118043 and SALK_095108 are shown on the map of the AtFAAH gene. Boxes represent exons, and lines represent introns and untranscribed flanking sequences. (B) Construct for constitutive expression of AtFAAH (35S::AtFAAH). NOS, nopaline synthase. (C) Semiquantitative RT-PCR analysis of transcript levels for AtFAAH knockouts and overexpressors. Total RNA was extracted from leaves of each homozygous AtFAAH mutant, overexpressing line and wild-type. The eIF4A-2 gene was used as control.
Fig. 7. Seedlings of AtFAAH overexpressors have higher fresh weights than vector controls. Three-day-old seedlings were transplanted onto fresh MS media, and seedling fresh weight was taken 8 days after transplant. Data show pooled fresh weight from 12 seedlings and five independent experiments.
Fig. 8. Effect of exogenous NAE18:2 on growth of Arabidopsis seedlings. (A) Arabidopsis seedlings exposed to 20 mM NAE18:2 for 3 days have significantly reduced organ growth. (B) Hypocotyl length of wild-type and AtFAAH knockouts exposed to 20 and 40 mM NAE18:2. Note that wild-type seedlings exhibit similar sensitivity to NAE18:2 as the knockouts. (C) Vector control and AtFAAH overexpressor (OE) seedlings exposed to NAE18:2 for 3 days. (D) Cotyledon area of wild-type and AtFAAH OE exposed to 20 and 40 mM NAE18:2. Note that AtFAAH OEs are more tolerant to NAE18:2 compared to vector controls and wild-type.
Table 1. Summary of phenotypic characteristics of WT, FAAH-overexpressing lines (2 and 11), and T-DNA mutants (SALK 095108 and 118043)
Phenotypic parameter | WT | OE2 | OE11 | SK095108 | SK118043 |
Seedling root length, 6-d old, mm† | 18.2 ± 0.7 | 23.8 ± 0.5*** | 22.5 ± 0.5*** | 18.5 ± 0.4 | 16.5 ± 0.5* |
Seedling hypocotyl length, 6-d old, mm† | 2.38 ± 0.04 | 3.37 ± 0.09*** | 3.30 ± 0.05*** | 2.44 ± 0.04 | 2.29 ± 0.06 |
Seedling cotyledon area, 6-d old, mm2† | 1.65 ± 0.08 | 2.20 ± 0.09*** | 2.27 ± 1.10*** | 1.45 ± 0.08 | 1.38 ± 0.08* |
Number of rosette leaves, 20-d old‡ | 5.6 ± 0.8 | 7.0 ± 0.7*** | 7.9 ± 1.0*** | 5.6 ± 1.6 | 5.2 ± 2.2 |
Number of rosette leaves, 28-d old‡ | 9.3 ± 1.4 | 12.4 ± 1.3*** | 12.5 ± 1.2*** | 9.2 ± 1.6 | 8.5 ± 1.1 |
Rosette diameter, 28-d old, cm‡ | 4.0 ± 1.0 | 4.7 ± 0.8** | 5.0 ± 0.7*** | 3.8 ± 0.7 | 3.2 ± 1.1* |
Number of days to bolting‡ | 39.3 ± 3.8 | 37.2 ± 3.6* | 32.6 ± 2.4*** | 38.8 ± 3.9 | 39.9 ± 4.3 |
Number of days first flower‡ | 43.0 ± 3.4 | 40.9 ± 3.0* | 36.8 ± 2.6*** | 41.6 ± 4.7 | 41.7 ± 3.5 |
Number of days to first silique‡ | 45.3 ± 3.0 | 43.3 ± 2.3* | 39.8 ± 2.6*** | 44.8 ± 3.8 | 44.5 ± 4.4 |
Height of inflorescence, 45 d-old, cm‡ | 8.9 ± 4.1 | 19.2 ± 7.0** | 27.0 ± 5.9*** | 10.3 ± 5.2 | 9.1 ± 4.6 |
Height of inflorescence, 52 d-old, cm‡ | 21.5 ± 7.6 | 35.7 ± 6.2*** | 42.1 ± 7.8*** | 23.7 ± 8.5 | 19.6 ± 7.5 |
Dry weight/plant, mg | 315 (n=35) | 334 (n=35) | 404 (n=33) | 255 (n=38) | 387 (n=44) |
Seed yield/plant, mg§ | 82.2 | 79.6 | 123 | 62.4 | 88.4 |
*P<0.01, **P< 0.0001, *** P<0.000001 (t-test versus WT).
†
Mean ± standard error.‡
Mean ± standard deviation.§
No difference in seed size or morphology, only in seed numbers.