Article Figures & Data
Figures
- Figure 1.
Scheme of the Chronological Progression of Principal Growth Stages in Arabidopsis.
Horizontal bars indicate the period during wild-type Col-0 plant development when the indicated trait can be used in growth stage determination. Numbers in parentheses correspond to principal growth stages listed in Table 1.
- Figure 2.
Arabidopsis Growth Stages.
(A) Stage 0.1, imbibition.
(B) Stage 0.5, radicle emergence.
(C) Stage 0.7, hypocotyl and cotyledons emerged from seed coat.
(D) Stage 1.0, cotyledons opened fully.
(E) Stage 1.02, two rosette leaves >1 mm in length.
(F) Stage 1.04, four rosette leaves >1 mm in length.
(G) Stage 1.10, ten rosette leaves >1 mm in length.
(H) Stage 5.10, first flower buds visible (indicated by arrow in inset).
(I) Stage 6.00, first flower open.
(J) Stage 6.50, midflowering.
(K) Stage 6.90, flowering complete.
(L) Stage 9.70, senescent and ready for seed harvest.
(A) to (F) were determined in the early analysis platform. (G) to (L) were determined in the soil-based platform.
- Figure 3.
Representative Data from Wild-Type Col-0 Plants.
(A) Number of rosette leaves >1 mm in length produced over time.
(B) Maximum rosette radius (i.e., length of the longest rosette leaf) over time.
(C) Plant height over time.
Arrows indicate the time at which growth stages 5.10, 6.00, and 6.50 occur. Data are given as averages ±sd for >300 individual plants. Days are given relative to date of sowing, including a 3-day stratification at 4°C to synchronize seed germination.
- Figure 4.
Growth Stage Progression for Wild-Type (Col-0) and Five Mutant Lines.
(A) Progression as determined in the plate-based early analysis platform.
(B) Progression as determined in the soil-based analysis platform.
Arrows define the time (days after sowing) at which Col-0 plants reached the growth stages indicated. Boxes represent the time elapsed between the occurrence of successive growth stages. Junctions between boxes of different shading indicate the occurrence of a growth stage. In (B), overlapping boxes for fae1-1 indicate that growth stage 5.10 was reached before stage 1.10. Days are given relative to date of sowing, including a 3-day stratificaton at 4°C to synchronize seed germination.
- Figure 5.
Detection of Phenotypic Differences between Mutants and Wild-Type Col-0 Plants.
(A) Comparison of leaf initiation between hls1-1 and Col-0. The number of leaves >1 mm were measured on day 14. Data are averages of 40 or >100 plants for hls1-1 and Col-0, respectively.
(B) Comparison of shoot growth between fae1-1 and Col-0. Total exposed leaf area was determined via a computerized analysis of digital images of intact rosettes (see Methods). Data are averages of 10 or >100 plants for fae1-1 and Col-0, respectively.
(C) Comparison of root growth between adg1-1 and Col-0. Both measurements were taken on day 14 (11 days after transfer to the growth chamber) and represent 20 seedlings grown on two plates.
(D) Comparison of silique area and the number of seed per half-silique between adg1-1 and Col-0. Silique area was determined via computerized analysis of digital images of mature filled siliques (see Methods). The number of seed per half-silique was observed after removal of the outer layer of one valve of a mature filled silique. Both measures were averaged from three siliques per plant and are reported as the average of 10 or >300 plants for adg1-1 and Col-0, respectively.
(E) Comparison of silique number and yield per plant between adg1-1 and Col-0. The final number of siliques per plant was determined after the completion of flower production (stage 6.90). Yield is reported as the desiccated mass (mg) of seed produced per plant. Data are averages and standard deviations of 10 or >300 plants for adg1-1 or Col-0, respectively.
(A) to (C) were obtained in the plate-based assay, and (D) and (E) were obtained in the soil-based assay.
Error bars indicate ±sd.
Tables
Col-0 Data Stage Description Daysa sd CVb Principal growth stage 0 Seed germination 0.10 Seed imbibition 3.0 NAc NA 0.50 Radicle emergence 4.3 0.4 10.3 0.7 Hypocotyl and cotyledon emergence 5.5 0.6 11.2 Principal growth stage 1 Leaf development 1.0 Cotyledons fully opened 6.0 0.5 8.5 1.02 2 rosette leaves >1 mm 10.3 0.6 5.8 1.04 4 rosette leaves >1 mm 14.4 0.5 3.4 Stage R6 More than 50% of the seedlings have primary roots ≥6 cm in length NDd ND ND a Average day from date of sowing, including a 3-day stratification at 4°C to synchronize germination.
b CV, coefficient of variation, calculated as (sd/days) × 100.
c NA, not applicable.
d ND, not determined (see text for details).
Col-0 Data Stage Description Daysa sd CVb Principal growth stage 1 Leaf development 1.02 2 rosette leaves >1 mm in length 12.5 1.3 10.7 1.03 3 rosette leaves >1 mm in length 15.9 1.5 9.5 1.04 4 rosette leaves >1 mm in length 16.5 1.6 9.8 1.05 5 rosette leaves >1 mm in length 17.7 1.8 10.2 1.06 6 rosette leaves >1 mm in length 18.4 1.8 9.8 1.07 7 rosette leaves >1 mm in length 19.4 2.2 11.1 1.08 8 rosette leaves >1 mm in length 20.0 2.2 11.2 1.09 9 rosette leaves >1 mm in length 21.1 2.3 10.8 1.10 10 rosette leaves >1 mm in length 21.6 2.3 10.9 1.11 11 rosette leaves >1 mm in length 22.2 2.5 11.2 1.12 12 rosette leaves >1 mm in length 23.3 2.6 11.3 1.13 13 rosette leaves >1 mm in length 24.8 3.2 12.8 1.14 14 rosette leaves >1 mm in length 25.5 2.6 10.2 Principal growth stage 3 Rosette growth 3.20 Rosette is 20% of final size 18.9 3.0 16.0 3.50 Rosette is 50% of final size 24.0 4.1 17.0 3.70 Rosette is 70% of final size 27.4 4.1 15.0 3.90 Rosette growth complete 29.3 3.5 12.0 Principal growth stage 5 Inflorescence emergence 5.10 First flower buds visible 26.0 3.5 13.3 Principal growth stage 6 Flower production 6.00 First flower open 31.8 3.6 13.3 6.10 10% of flowers to be produced have opened 35.9 4.9 13.6 6.30 30% of flowers to be produced have opened 40.1 4.9 12.3 6.50 50% of flowers to be produced have opened 43.5 4.9 11.2 6.90 Flowering complete 49.4 5.8 11.7 Principal growth stage 8 Silique ripening 8.00 First silique shattered 48.0 4.5 9.3 Principal growth stage 9 Senescence 9.70 Senescence complete; ready for seed harvest NDc ND ND a Average day from date of sowing, including a 3-day stratification at 4°C to synchronize germination.
b CV, coefficient of variation, calculated as (sd/days) × 100.
c ND, not determined (see text for details).
First-Phase Measurements Measurement/Query Growth Stage Defined Has radicle emergence been reached or passed? Stage 0.5 Are the hypocotyl and cotyledons visible? Stage 0.7 Have the cotyledons opened fully? Stage 1.0 Number of rosette leaves >1 mm Principal growth stage 1 Length of primary root ≥6 cm Stage R6 First-Phase Measurements Measurement/Query Growth Stage Defined Rosette radius Principal growth stage 3 Are flower buds visible? Stage 5.10 Is first flower open? Stage 6.00 Length of stem Stage 6.50a Number of open flowers Principal growth stage 6 Number of senescent flowers Principal growth stage 6 Number of filled siliques Principal growth stages 6 and 7 Number of shattered siliques Principal growth stage 8 Is flower production complete? Stage 6.90 Col-0 Data Measurement Unit Growth Stage Average sd CVa Number of rosette leaves Count 1.0b 3.3 0.5 13.7 Length of primary root mm 1.0b 45.2 4.1 9.0 Number of secondary roots Count R6c 10.5 1.4 13.2 Rosette, total exposed leaf area mm2 R6c 22.2 2.6 11.9 Rosette, perimeter mm R6c 42.1 5.2 12.2 Rosette, sd of radius None R6c 39.2 3.1 7.9 Rosette, major axis mm R6c 7.9 0.7 9.1 Rosette, minor axis mm R6c 5.9 0.6 9.5 Rosette, eccentricity None R6c 0.63 0.05 7.2 a CV, coefficient of variation, calculated as (sd/days) × 100.
b Data collection initiated at stage 1.0 and continued until the end of the experiment.
c R6 or 14 days after sowing, whichever comes first.
Col-0 Data Measurement Unit Growth Stage Average sd CVb Number of cotyledons Count 1.04 2.0 0.1 5.0 Rosette, total exposed leaf area mm2 1.10 580.0 202.2 34.9 Rosette, perimeter mm 1.10 418.0 119.1 28.5 Rosette, sd of radius None 1.10 45.7 3.2 7.0 Rosette, major axis mm 1.10 40.4 8.0 19.8 Rosette, minor axis mm 1.10 34.6 7.5 21.7 Rosette, eccentricity None 1.10 0.5 0.1 20.0 Rosette, total exposed leaf area mm2 6.00 3225.0 1088.3 33.7 Rosette, perimeter mm 6.00 808.1 181.3 22.4 Rosette, sd of radius None 6.00 36.7 3.5 9.5 Rosette, major axis mm 6.00 82.3 15.3 18.6 Rosette, minor axis mm 6.00 73.1 13.4 18.3 Rosette, eccentricity None 6.00 0.4 0.1 25.0 Rosette, dry weight mg 6.00 117.4 45.9 39.1 Number of stem branches on main bolt Count 6.50 3.4 0.6 17.6 Number of side bolts >1 cm Count 6.50 4.2 1.2 28.6 Length of peduncle of second flower on main bolt mm 6.50 11.5 1.6 13.9 Distance across face of open flower mm 6.50 3.9 0.3 7.7 Sepal length mm 6.50 2.2 0.2 9.1 Pollen grain, area μm2 6.50 589.0 132.0 22.4 Pollen grain, perimeter μm 6.50 114.5 13.0 11.2 Pollen grain, sd of radius None 6.50 9.1 2.5 27.5 Pollen grain, major axis μm 6.50 30.6 3.3 10.8 Pollen grain, minor axis μm 6.50 24.3 3.0 12.3 Pollen grain, eccentricity None 6.50 0.6 0.1 16.7 Silique, area mm2 6.50 10.6 1.9 17.9 Silique, perimeter mm 6.50 40.9 6.1 14.9 Silique, sd of radius None 6.50 55.8 0.5 0.9 Silique, major axis mm 6.50 17.2 1.7 9.9 Silique, minor axis mm 6.50 1.2 0.2 16.7 Silique, eccentricity None 6.50 1.0 0.0 0.0 Total number of seeds per silique valve Count 6.50 29.9 2.8 9.4 Number of abnormal seeds per silique valve Count 6.50 0.2 0.4 200 Dry weight of stem mg 6.50 188.8 39.3 20.8 Dry weight of rosette mg 6.50 163.7 52.0 31.8 Total number of siliques Count 6.90 160.4 60.7 37.8 Seed, area mm2 9.70 0.14 0.01 7.1 Seed, perimeter mm 9.70 1.95 0.04 2.1 Seed, sd of radius None 9.70 16.92 0.94 5.6 Seed, major axis mm 9.70 0.53 0.03 5.7 Seed, minor axis mm 9.70 0.33 0.02 6.1 Seed , eccentricity None 9.70 0.78 0.02 2.6 Seed yield per plant (desiccated) mg 9.70 127.9 52.7 41.2 a Used to define the working definition of stage 6.50 as described in the text.
b CV, coefficient of variation, calculated as (sd/days) × 100.
