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Dynamic morphologies of pollen plastids visualised by vegetative-specific FtsZ1–GFP in Arabidopsis thaliana

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Abstract

The behaviour and multiplication of pollen plastids have remained elusive despite their crucial involvement in cytoplasmic inheritance. Here, we present live images of plastids in pollen grains and growing tubes from transgenic Arabidopsis thaliana lines expressing stroma-localised FtsZ1–green-fluorescent protein fusion in a vegetative cell-specific manner. Vegetative cells in mature pollen contained a morphologically heterogeneous population of round to ellipsoidal plastids, whilst those in late-developing (maturing) pollen included plastids that could have one or two constriction sites. Furthermore, plastids in pollen tubes exhibited remarkable tubulation, stromule (stroma-filled tubule) extension, and back-and-forth movement along the direction of tube growth. Plastid division, which involves the FtsZ1 ring, was rarely observed in mature pollen grains.

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Acknowledgements

The authors are grateful to Prof. Danny J. Schnell (University of Massachusetts at Amherst, USA) for the gift of antiserum against Arabidopsis Tic110, Dr. Yasuo Niwa (University of Shizuoka, Japan) for the original GFP vector, Arabidopsis Biological Resource Center (Ohio State University, USA) for providing the arc11 seeds and Prof. Rachel M. Leech and Ms. Joanne L. Marrison (University of York, UK) for donating them. We also thank Dr. Yukihisa Shimada (RIKEN Plant Science Center, Japan), Dr. Takeshi Nakano, Ms. Sumie Ohbu (RIKEN), Mr. Tadasuke Chiba (KS Olympus, Japan) and the facility of RIKEN Radioactive Isotope Beam Factory for supporting this study. This work was supported by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan (No. 17780077 to M.T.F., No. 20657015 to R.D.I.) and the Agricultural Chemical Research Foundation (to M.T.F. and R.D.I.).

Conflict of Interest

The authors declare that they have no conflict of interest.

Author information

Correspondence to Makoto T. Fujiwara.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Time-lapse observation of FtsZ1–GFP in a germinating pollen grain of the wild-type. Fluorescence images were taken for 120 s at 4-s intervals. Germination has occurred at the left part of grain; 120 s corresponds to 3.0 s in this movie. The pollen sample is the same as that in Fig. 4d. (MPG 380 kb)

Time-lapse observation of FtsZ1–GFP in germinated wild-type pollen. Fluorescence images were taken for 116 s at 4-s intervals. 116 s correspond to 2.9 s in this movie. The pollen sample is the same as that in Fig. 4f. (MPG 288 kb)

Z-series images of FtsZ1–GFP in a mature pollen grain of arc11. Fluorescence images taken for 52 s at 4-s intervals are shown in this movie. The pollen sample is the same as that in Fig. 5b. (MPG 232 kb)

Movie S1

Time-lapse observation of FtsZ1–GFP in a mature wild-type pollen grain. Fluorescence images were taken for 60 s at 4-s intervals; 60 s corresponds to 1.5 s in this movie. Note that pollen was incubated on in vitro germination medium before observation. (MPG 194 kb)

Movie S2

Time-lapse observation of FtsZ1–GFP in a mature wild-type pollen grain. Fluorescence images were taken for 100 s at 4-s intervals; 100 s corresponds to 2.5 s in this movie. Note that the pollen sample is same with that of Fig. 2h and was not incubated before observation. (MPG 210 kb)

Movie S3

Time-lapse observation of a mature wild-type pollen grain. DIC images were taken for 76 s at 4-s intervals; 76 s corresponds to 1.9 s in this movie. Note that pollen was incubated on in vitro germination medium before observation. (MPG 396 kb)

Movie S4

Z-series images of FtsZ1–GFP in a maturing pollen grain of the wild type. Fluorescence images taken for 60 s at 4-s intervals are shown in this movie. (MPG 142 kb)

Movie S5

Time-lapse observation of FtsZ1–GFP in maturing wild-type pollen grain. Fluorescence images were taken for 120 s at 4-s intervals; 120 s corresponds to 3.0 s in this movie. The pollen sample is the same as that in Fig. 3c. (MPG 286 kb)

Movie S6

Time-lapse observation of FtsZ1–GFP in a germinating wild-type pollen grain. Fluorescence images were taken for 80 s at 4-s intervals; 80 s corresponds to 2.0 s in this movie. Germination is just about to occur at the left part of the grain. The pollen sample is the same as that in Fig. 4b. (MPG 164 kb)

Movie S7

Time-lapse observation of FtsZ1–GFP in a germinating pollen grain of the wild-type. Fluorescence images were taken for 120 s at 4-s intervals. Germination has occurred at the left part of grain; 120 s corresponds to 3.0 s in this movie. The pollen sample is the same as that in Fig. 4d. (MPG 380 kb)

Movie S8

Time-lapse observation of a germinating wild-type pollen tube. DIC images were taken for 132 s at 4-s intervals; 132 s corresponds to 3.3 s in this movie. (MPG 268 kb)

Movie S9

Time-lapse observation of FtsZ1–GFP in germinated wild-type pollen. Fluorescence images were taken for 116 s at 4-s intervals. 116 s correspond to 2.9 s in this movie. The pollen sample is the same as that in Fig. 4f. (MPG 288 kb)

Movie S10

Time-lapse observation of the growing tip of the wild-type pollen tube. DIC images were taken for 128 s at 4-s intervals; 128 s correspond to 3.2 s in this movie. (MPG 348 kb)

Movie S11

Time-lapse observation of FtsZ1–GFP in the pollen tube of the wild type. Fluorescence images were taken for 136 s at 4-s intervals. Tip growth (the arrow shows the tip point) was in the right-to-left direction; 136 s correspond to 3.4 s in this movie. The pollen sample is the same as that in Fig. 4j. (MPG 1326 kb)

Movie S12

Time-lapse observation of FtsZ1–GFP in a wild-type pollen tube. Fluorescence images were taken for 195 s at 5-s intervals. Tip growth was in the left-to-right direction; 195 s corresponds to 3.9 s in this movie. (MPG 358 kb)

Movie S13

Z-series images of FtsZ1–GFP in a mature pollen grain of arc11. Fluorescence images taken for 52 s at 4-s intervals are shown in this movie. The pollen sample is the same as that in Fig. 5b. (MPG 232 kb)

Movie S14

Time-lapse observation of FtsZ1–GFP in a mature pollen grain of arc11. Fluorescence images were taken for 51 s at 3-s intervals; 51 s corresponds to 1.7 s in this movie. (MPG 124 kb)

Movie S15

Time-lapse observation of FtsZ1–GFP in a pollen tube of arc11. Fluorescence images were taken for 72 s at 4-s intervals. A whole picture of germinated pollen is presented; 72 s corresponds to 1.5 s in this movie. (MPG 402 kb)

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Fujiwara, M.T., Hashimoto, H., Kazama, Y. et al. Dynamic morphologies of pollen plastids visualised by vegetative-specific FtsZ1–GFP in Arabidopsis thaliana . Protoplasma 242, 19–33 (2010). https://doi.org/10.1007/s00709-010-0119-7

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Keywords

  • Arabidopsis thaliana
  • FtsZ ring
  • Leucoplast
  • Plastid division
  • Stromule