Figure 5.
Loss of function of ADF7 and ADF10 has differential effects on vesicle trafficking in pollen tubes. (A) Images of pollen tubes harboring YFP-RabA4b. Yellow dots mark the V-shaped region of vesicle accumulation in WT, adf7, and adf10 pollen tubes. Bar = 5 μm. (B) Quantification of the average angles of the V-shaped area of vesicle accumulation in the apical region of WT adf7 and adf10 pollen tubes. Values represent mean ± SD. n > 25. **P < 0.01 (unpaired two-tailed Student’s t test). (C) Quantification of the distance of the top of the V shape from the bottom of the V (depth of V) in the pollen tube. Values represent mean ± SD, n > 15. **P < 0.01 (unpaired two-tailed Student’s t test). (D) Quantification of the distance of the top of the V from the tip of the pollen tube. Values represent mean ± SD, n > 15. **P < 0.01 (unpaired two-tailed Student’s t test). (E) Time-lapse images of YFP-RabA4b fluorescence recovery after photobleaching at the tip of pollen tubes. The red box indicates the photobleached region. Bar = 5 μm. (F) The fitted fluorescence recovery curves of Col-0 (WT), adf7, and adf10 were obtained by measuring the fluorescence intensities in the 5 μm region at the tube tips. More than 20 pollen tubes for each genotype were included. (G) YFP-RabA4b fluorescence recovers much more slowly after photobleaching in adf10 pollen tube tips than in Col-0 (WT) and adf7 pollen tubes. Error bars indicate SE (n > 20). ***P < 0.001 by all-pairwise one-way ANOVA test. (H) Transverse sections of the pollen tubes are shown in A. The distances from the extreme tip are indicated in the images. Bar = 5 μm. (I) Plot of the fluorescence intensity of YFP-RabA4b at the pollen tube tip within the band perpendicular to the growth axis of pollen tubes. (J) Plot of the fluorescence intensity of YFP-RabA4b at the pollen tube tip within the band that is parallel to the growth axis of pollen tubes. (K) Kymographs of RabA4b-positive particles in the first 30 s after photobleaching. The upper panel shows the gray-shaded region of the photobleached pollen tube which was included to build the kymograph. White arrowheads indicate the regions where RabA4b-positive particles are accumulated, and yellow triangles indicate the extreme apical region of vesicle accumulation. Bar = 2 μm.

Loss of function of ADF7 and ADF10 has differential effects on vesicle trafficking in pollen tubes. (A) Images of pollen tubes harboring YFP-RabA4b. Yellow dots mark the V-shaped region of vesicle accumulation in WT, adf7, and adf10 pollen tubes. Bar = 5 μm. (B) Quantification of the average angles of the V-shaped area of vesicle accumulation in the apical region of WT adf7 and adf10 pollen tubes. Values represent mean ± SD. n > 25. **P < 0.01 (unpaired two-tailed Student’s t test). (C) Quantification of the distance of the top of the V shape from the bottom of the V (depth of V) in the pollen tube. Values represent mean ± SD, n > 15. **P < 0.01 (unpaired two-tailed Student’s t test). (D) Quantification of the distance of the top of the V from the tip of the pollen tube. Values represent mean ± SD, n > 15. **P < 0.01 (unpaired two-tailed Student’s t test). (E) Time-lapse images of YFP-RabA4b fluorescence recovery after photobleaching at the tip of pollen tubes. The red box indicates the photobleached region. Bar = 5 μm. (F) The fitted fluorescence recovery curves of Col-0 (WT), adf7, and adf10 were obtained by measuring the fluorescence intensities in the 5 μm region at the tube tips. More than 20 pollen tubes for each genotype were included. (G) YFP-RabA4b fluorescence recovers much more slowly after photobleaching in adf10 pollen tube tips than in Col-0 (WT) and adf7 pollen tubes. Error bars indicate SE (n > 20). ***P < 0.001 by all-pairwise one-way ANOVA test. (H) Transverse sections of the pollen tubes are shown in A. The distances from the extreme tip are indicated in the images. Bar = 5 μm. (I) Plot of the fluorescence intensity of YFP-RabA4b at the pollen tube tip within the band perpendicular to the growth axis of pollen tubes. (J) Plot of the fluorescence intensity of YFP-RabA4b at the pollen tube tip within the band that is parallel to the growth axis of pollen tubes. (K) Kymographs of RabA4b-positive particles in the first 30 s after photobleaching. The upper panel shows the gray-shaded region of the photobleached pollen tube which was included to build the kymograph. White arrowheads indicate the regions where RabA4b-positive particles are accumulated, and yellow triangles indicate the extreme apical region of vesicle accumulation. Bar = 2 μm.

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