Figure 3.

Absence of cellulose dependent florescence arising between columella striations in mutants involving CESA5. Analysis of Calcofluor white stained seeds showed lack of florescence between striations of fluorescence arising from regions coincident with columella in the mutant seeds involving CESA5 (cesa5–1 and cesa2–1cesa5–1cesa9–1) compared with wild type and other mutant seeds. The intensity and length of the florescence halo surrounding the seed was reduced in cesa2–1, cesa9–1 and cesa2–1cesa9–1 compared with wild type. To visualize the calcofluor florescence, wild type and mutant seeds were stained with Calcofluor white for 5 min and observed under laser scanning confocal microscopy. Three dimensional reconstructions were made by taking images on a Z-series at 2 μm intervals and threading them into an animation. The images represent three different angles and illustrate the uniformity of the phenotype across a three dimensional plane. Scale bar = 250 μm.