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Mechanism of action of nitrous oxide gas applied as a polyploidizing agent during meiosis in lilies

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Abstract

Nitrous oxide gas (N2O) can be used to produce polyploid plants, but the mechanism of action is unknown. The actin and microtubule cytoskeleton was observed in N2O-treated microsporocytes of Lilium spp ‘Asiatic hybrid lilies’ using fluorescence microscopy after staining with DAPI, FITC-conjugated tubulin antibody, and phalloidin-conjugated Alexa Fluor 546. Additionally, microsporocytes of L. longiflorum were observed with acetocarmine staining following N2O treatment. A typical metaphase I microtubule distribution was observed in control microsporocytes. After treatment with N2O for 24 h, microtubules were effectively depolymerized; this prevented chromosomes from moving to the poles, resulting in chromosome retention in the center of N2O-treated cells. Cell plate formation took place without delay, however, yielding one daughter cell with a diploid genome and another daughter without chromosomes. In addition, N2O treatment often induced micronuclei due to aberrant chromosome separation during cytokinesis. Actin filaments in microsporocytes are insensitive to N2O. These findings indicate that N2O mediates polyploidization by inhibiting microtubule polymerization, but not actin filament formation, during microsporocyte meiosis.

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Acknowledgments

The authors gratefully acknowledge the helpful advice of Professor K. Takeno of Niigata University. This research was supported by a Grant-in-Aid for Scientific Research (No. 19580024) from the Ministry of Education, Culture, Sports, Science and Technology, Japan and by a Research Project for Utilizing Advanced Technology in Agriculture, Forestry and Fisheries from the Ministry of Agriculture, Forestry and Fisheries of Japan.

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  1. Masako Akutsu

    Present address: National Agricultural Research Center for Hokkaido Region, 1 Hitsujigaoka, Toyohiraku, Sapporo, Hokkaido, 062-8555, Japan

Authors and Affiliations

  1. Faculty of Agriculture, Niigata University, Ikarashi, Niigata, 950-2181, Japan

    Satomi Kitamura, Masako Akutsu & Keiichi Okazaki

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  1. Satomi Kitamura
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  2. Masako Akutsu
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  3. Keiichi Okazaki
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Correspondence to Keiichi Okazaki.

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Communicated by Hugh Dickinson.

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Kitamura, S., Akutsu, M. & Okazaki, K. Mechanism of action of nitrous oxide gas applied as a polyploidizing agent during meiosis in lilies. Sex Plant Reprod 22, 9–14 (2009). https://doi.org/10.1007/s00497-008-0084-x

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  • DOI: https://doi.org/10.1007/s00497-008-0084-x

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