Article Figures & Data
Figures
- Figure 1.
KNAT6 Is Expressed in Boundaries.
(A) Expression of pKNAT6-GUS was detected at the torpedo stage. The embryos were cleared.
(B) Longitudinal section through a globular embryo. The KNAT6 mRNA was not detected.
(C) Longitudinal section through a triangular embryo showing KNAT6 mRNA accumulation in the presumptive SAM.
(D) and (E) Longitudinal sections through late-torpedo (D) and mature (E) embryos showing KNAT6 mRNA accumulation in the boundaries between the SAM and the cotyledons.
(F) Transverse section through a mature embryo showing KNAT6 mRNA accumulation in a region surrounding the SAM. This region corresponds to the boundaries between the SAM (S) and the cotyledons (C) and the boundaries between the cotyledons.
Bars = 50 μm.
- Figure 2.
Characterization of the Loss-of-Function knat6 and knat2 Mutant Alleles.
(A) RT-PCR analysis using KNAT6 gene-specific primers. After 38 cycles, the PCR products were transferred to a nylon membrane and hybridized with a 33P-labeled KNAT6 probe. Amplification occurred in wild-type seedlings (lane 1) but not in knat6-1 and knat6-2 alleles (lanes 2 and 3, respectively), indicating that both alleles are null. Control amplification of APT1 indicates that equal amounts of cDNA were present in each sample.
(B) RT-PCR analysis using KNAT2 gene-specific primers. After 40 cycles, the PCR products were transferred to a nylon membrane and hybridized with a 33P-labeled KNAT2 probe. Amplification occurred in wild-type seedlings (lane 1) but not in the knat2-5 allele (lane 2), indicating that the knat2-5 allele is null. Control amplification of APT1 indicates that equal amounts of cDNA were present in each sample.
- Figure 3.
Phenotype of the knat6 stm Double Mutant.
All seedlings shown are 8 d old. Bars = 100 μm.
(A) Wild-type seedling. The SAM gives rise to leaf primordia.
(B) stm-2 seedling.
(C) knat6-1 stm-2 seedling showing a fusion that extends to the blade of the cotyledons.
(D) Longitudinal section through a wild-type vegetative meristem.
(E) Longitudinal section through a stm-2 seedling. A primordium is visible between the cotyledons.
(F) Longitudinal section through a knat6-1 stm-2 seedling. The cells are large and vacuolated.
- Figure 4.
WET368 GUS Expression in knat6-1 Single Mutant and knat6-1 stm-2 Double Mutant.
(A) knat6-1/knat6-1 mature embryo showing GUS activity in the SAM boundaries. This activity was identical in the wild-type embryo.
(B) knat6-1/knat6-1 stm-2/stm-2 mature embryo showing a dramatic reduction of GUS activity.
(C) knat6-1/knat6-1 8-d-old seedling showing GUS activity in the meristem. Seedlings were cleared.
(D) stm-2/stm-2 8-d-old seedling exhibiting GUS activity.
(E) knat6-1/knat6-1 stm-2/stm-2 8-d-old seedling exhibiting no GUS activity.
Bars = 50 μm.
- Figure 5.
Triple Mutants of knat6-1 stm-2 cuc1-1, knat6-1 stm-2 cuc2, and knat6-1 stm-2 cuc3-2.
Eight-day-old seedlings of the wild type ([A] and [E]), cuc1-1 cuc2 ([B] and [F]), cuc1-1 cuc3-2 ([C] and [G]), cuc2 cuc3-2 ([D] and [H]), knat6-1 stm-2 ([I] and [M]), knat6-1 stm-2 cuc1-1 ([J] and [N]), knat6-1 stm-2 cuc2 ([K] and [O]), and knat6-1 stm-2 cuc3-2 ([L] and [P]).
- Figure 6.
pKNAT6-GUS Expression in Mutant Backgrounds.
(A) Wild-type mature embryo showing GUS activity in the SAM boundaries and in the tips of cotyledons. The embryo was cleared.
(B) cuc1-1/cuc1-1 cuc2/cuc2 double mutant mature embryo showing no GUS activity in the SAM. The expression in the cotyledon tips is maintained.
(C) stm-5/stm-5 mature embryo showing a reduction of GUS activity in the SAM. This activity is reduced to two faint spots. The expression in the cotyledon tips is maintained.
(D) Wild-type 8-d-old cleared seedling showing GUS activity in the meristem.
(E) cuc1-1/cuc1-1 cuc2/cuc2 seedling showing no GUS activity.
(F) stm-5/stm-5 seedling exhibiting no GUS activity.
Bars = 50 μm.
Additional Files
Supplemental Data
Files in this Data Supplement:
- Supplemental Figure 1 (Revised) - pKNAT6-GUS expression. (A) Histolocalization of KNAT6-GUS activity in the SAM. Longitudinal section through a vegetative meristem. The expression of pKNAT6-GUS was restricted to the boundaries of the meristem and the stipules. (B) The expression of pKNAT6-GUS was localized in the vasculature elements of the mature root. (C) The KNAT6-GUS activity was detected in the lateral roots. Bars = 50 μM
- Supplemental Figure 2 - KNAT2 expression in embryo. (A) Longitudinal section through a late globular stage embryo showing KNAT2 mRNA accumulation. (B) Longitudinal section through a heart stage embryo showing KNAT2 mRNA accumulation in the center of the embryo. (C) Longitudinal section through a mature embryo showing KNAT2 mRNA accumulation in the SAM. (D) Longitudinal section through a mature embryo showing KNAT2-GUS activity in the SAM. Bars = 50 μM
- Supplemental Figure 3 - Phenotype of the knat2-5 stm-2and the knat2-5 knat6-1 stm-2 mutants. (A and D) knat2-5 seedlings. (B and E) knat2-5 stm-2 seedlings. (C and D) knat2-5 stm-2 knat6-1 seedlings.
