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First published online 24 January 2007
doi: 10.1242/dev.02774

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Plants expressing a miR164-resistant CUC2 gene reveal the importance of post-meristematic maintenance of phyllotaxy in Arabidopsis

Laboratoire de Biologie Cellulaire UR501, Institut J. P. Bourgin, INRA, F-78000 Versailles, France.

View larger version (54K): [in this window] [in a new window]   Fig. 1. Phyllotactic pattern in CUC2g-m4 and wild type. (A) Axillary region. An additional leaf develops in the axils of CUC2g-m4 plants (arrow). (B) Frequency of the extra leaf phenotype in independent transgenic lines. (C) Strong CUC2g-m4 plants with an extreme reduction of the inflorescence (arrow) and of flowers with long pedicels (arrowhead). (D) Inflorescence of wild-type and representative CUC2g-m4 plants showing an abnormal phyllotactic pattern. (E) Close-up of D. (F) Detailed analysis of the phyllotactic pattern in wild-type and CUC2g-m4 plants (WS background, see Fig. S2 in the supplementary material for Col). Left: distribution of divergence angle between two successive flowers along the stem. Percentages of total divergence angles measured (n) that fell into 12 classes, each of 30°, and the averages (av) are shown. Right: distribution of internode length between two successive flowers along the stem. Percentages of total internode lengths that fell into classes of 3 mm each are shown. Insets show a finer distribution of internodes that were less than 3 mm long, in classes of 0.5 mm. Notice that the scale is different for the insets.

View larger version (24K): [in this window] [in a new window]   Fig. 2. Meristems and meristematic phyllotaxy are unaltered in CUC2g-m4 plants. (A) Cleared inflorescence meristems of wild type and CUC2g-m4 (Col background). (B) Meristem width of wild type and CUC2g-m4 in WS and Col backgrounds. Number of individuals measured (n) and standard deviations (bars) are indicated. (C) Expression of the auxin response sensor DR5:GFP in wild type and CUC2g-m4 (WS background). (D) Meristematic phyllotaxy in wild type and CUC2g-m4 (WS background; see Fig. S2 in the supplementary material for Col). Percentages of total measurements (n) of divergence angle between two successive flowers in the meristem falling into twelve 30° classes and averages (av) are shown. Scale bars: 10 µm in A; 100 µm in C.

View larger version (17K): [in this window] [in a new window]   Fig. 3. Short internodes in CUC2g-m4 result from a post-meristematic reduction of growth due to a combination of fewer and smaller cells. (A) Developmental series of flowers along the stem. Flowers are ordered from the top (left) to the bottom (right) of the inflorescence stem. Not all of the flowers of a CUC2g-m4 cluster (underlined) are at the same stage. (B) SEM views of the early phases of inflorescence development. Inflorescences were partially dissected to reveal the internodes. (C) Epidermal cell length and number of wild-type (10 mm long), short CUC2g-m4 (less than 1 mm long, yellow bars) or long CUC2g-m4 (30 mm long, red bars) internodes. Cell numbers per internode were calculated based on cell and internode lengths. Number of individuals measured (n) and standard deviations (bars) are indicated. (D) Epidermal cell size was measured on either the inside or the outside of twisted CUC2g-m4 stems. Scale bar: 0.5 mm in B.

View larger version (135K): [in this window] [in a new window]   Fig. 4. CUC2 in situ hybridisation on longitudinal sections of inflorescence apices. (A) Wild type. (B,C,D) CUC2g-m4 transgenic plants. (C) Twisted stem with CUC2 expression on the inner side. (D) Close-up of the inner and outer sides of C. Arrows point to the lowest axils with detectable CUC2 expression. Arrowheads indicate ectopic CUC2 in the internodes. Scale bars: 100 µm.