Fig. 6: Cytoplasmic streaming impacts on VN movement in pollen tubes. | Nature Communications

Fig. 6: Cytoplasmic streaming impacts on VN movement in pollen tubes.

From: Distinct functions of microtubules and actin filaments in the transportation of the male germ unit in pollen

Fig. 6

a–c Time-lapse images of H2B-GFP in pollen tubes when VN moves forward (a), backward (b), or at pause (c). Black arrows indicate the vector distribution of the velocity of vesicles. Arrowheads indicate the tube growth direction. Magenta straight lines indicate the anisotropy of vesicle trafficking. Bars, 10 μm. d Quantification of the anisotropy of vesicle trafficking. Forward (n = 186), Pause (n = 154), and Backward (n = 177) from 10 cells. e Time-lapse images of Lifeact-mCherry and H2B-GFP in the germinating pollen grains. “0 s” sets at 10 min after the incubation on medium. Bar, 5 μm. f Trajectory analysis of VN movement in DMSO-, LatB- or PBP-treated pollen grains. Bars, 5 μm. g Quantification of the maximum distance of VN movement in DMSO, LatB, or PBP treatment. DMSO (n = 19), LatB (n = 18) and PBP (n = 18). h Time-lapse images of VN (green) and SCs (magenta) in DMSO-, LatB- or PBP-treated pollen tubes. Bars, 20 μm. i Kymograph analysis of (h). j Quantification of the velocity of vesicle trafficking in DMSO and PBP treatment. n = 36. k Quantification of VN’s movement velocity in DMSO and PBP treatment. Velocity of VN forward movement, WT (n = 310), 0.5 μM PBP (n = 135), 0.625 μM PBP (n = 116), 0.75 μM PBP (n = 44). Velocity of VN backward movement, WT (n = 63), 0.5 μM PBP (n = 56), 0.625 μM PBP (n = 34), 0.75 μM PBP (n = 22). l Quantification of the velocity of vesicle trafficking in WT and myosin mutants pollen tubes. WT (n = 104), myo11c1 (n = 113), myo11c2 (n = 103), myo11c1 myo11c2 (n = 103). m Quantification of VN’s movement velocity in WT and myosin mutants pollen tubes. Velocity of VN forward movement, WT (n = 704), myo11c1 (n = 691), myo11c2 (n = 735), myo11c1 myo11c2 (n = 690).Velocity of VN backward movement, WT (n = 170), myo11c1 (n = 140), myo11c2 (n = 107), myo11c1 myo11c2 (n = 134). n Quantification of the distance of VN to the tip of WT and myosin mutants. WT (n = 792), myo11c1 (n = 812), myo11c2 (n = 863), myo11c1 myo11c2 (n = 800). n from 10 cells for each treatment in (j), (k); n from 10 cells for each genotype in (l), (m), (n). Values are mean ± SD; *P < 0.05, **P < 0.01, ***P < 0.001, ns, not significant; two-tailed Student’s t-test in (d), (g), (j–n). Source data are provided as a Source Data file.

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