Hoechst-tagged Fluorescein Diacetate for the Fluorescence Imaging-based Assessment of Stomatal Dynamics in Arabidopsis thaliana

Yousuke Takaoka; Saki Miyagawa; Akinobu Nakamura; Syusuke Egoshi; Shinya Tsukiji; Minoru Ueda

doi:10.1038/s41598-020-62239-w

Hoechst-tagged Fluorescein Diacetate for the Fluorescence Imaging-based Assessment of Stomatal Dynamics in Arabidopsis thaliana

Sci Rep. 2020 Mar 24;10(1):5333. doi: 10.1038/s41598-020-62239-w.

Authors

Yousuke Takaoka^{1

2}, Saki Miyagawa³, Akinobu Nakamura⁴, Syusuke Egoshi¹, Shinya Tsukiji^{5

6

7}, Minoru Ueda^{8

9}

Affiliations

¹ Department of Chemistry, Graduate School of Science, Tohoku University, Sendai, 980-8578, Japan.
² Precursory Research for Embryonic Science and Technology (PREST), Japan Science and Technology Agency, 5 Sanbancho, Chiyoda-ku, Tokyo, 102-0075, Japan.
³ Department of Molecular and Chemical Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, 980-8578, Japan.
⁴ Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya, 466-8555, Japan.
⁵ Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya, 466-8555, Japan. stsukiji@nitech.ac.jp.
⁶ Department of Nanopharmaceutical Sciences, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya, 466-8555, Japan. stsukiji@nitech.ac.jp.
⁷ Frontier Research Institute for Materials Science (FRIMS), Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya, 466-8555, Japan. stsukiji@nitech.ac.jp.
⁸ Department of Chemistry, Graduate School of Science, Tohoku University, Sendai, 980-8578, Japan. minoru.ueda.d2@tohoku.ac.jp.
⁹ Department of Molecular and Chemical Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, 980-8578, Japan. minoru.ueda.d2@tohoku.ac.jp.

Abstract

In plants, stomata regulate water loss through transpiration for plant growth and survival in response to various environmental stressors; and simple methods to assess stomatal dynamics are needed for physiological studies. Herein, we report a fluorescence-imaging-based method using fluorescein diacetate tagged with Hoechst 33342, a nuclear staining chemical probe (HoeAc₂Fl) for the qualitative assessment of stomatal dynamics. In our method, the stomatal movement is inferred by simple monitoring of the fluorescence intensity in the nucleus of the stomata.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arabidopsis / metabolism
Arabidopsis Proteins / genetics
Carbon Dioxide / metabolism
Fluoresceins / chemistry*
Fluorescence
Fluorescent Dyes / chemistry
Gene Expression Regulation, Plant / genetics
Optical Imaging / methods*
Plant Leaves / metabolism
Plant Stomata / metabolism
Plant Stomata / physiology*
Plant Transpiration / physiology
Water / metabolism

Substances

Arabidopsis Proteins
Fluoresceins
Fluorescent Dyes
Water
Carbon Dioxide
diacetylfluorescein