A new application of the SFDA-staining method to assessment of the freezing tolerance in leaves of alpine plants

• Main Authors: Wataru Yamori, Hiroyuki Kogami, Yoshitaka Yoshimura, Takashi Tsuji, Takehiro Masuzawa
• Format: Report
• Language: English
• Published: Department of Biology and Geosciences, Faculty of Science, Shizuoka University 2006
• Subjects: alpine plants; FDA (fluorescein diacetate); freezing tolerance; SFDA (5- (6-) sulfofluorescein diacetate); survival ratio; Polar bioscience
• Online Access: https://nipr.repo.nii.ac.jp/?action=repository_uri&item_id=6262; http://id.nii.ac.jp/1291/00006262/; https://nipr.repo.nii.ac.jp/?action=repository_action_common_download&item_id=6262&item_no=1&attribute_id=18&file_no=1

For the first time, this study used 5- (6-) sulfofluorescein diacetate (SFDA), a fluorescent product in plant cells converted by esterase activity to fluorescein-5- (and 6-) sulfonic acid (FSA), to assess the freezing tolerance of leaf cells. We were able to readily distinguish living and dead cells, and detect differences in freezing tolerance among five alpine plants using the SFDA-staining method. We also compared this method with two conventional methods, the electrolyte leakage test and fluorescein diacetate (FDA) staining method. The electrolyte leakage test often over- or underestimated freezing injury. With the uninjured control samples, the FDA-staining method failed to stain all leaf cells, while the SFDA-staining method stained almost 100%. From these results, we concluded that SFDA-staining is a more convenient, accurate and reproducible method for analyses of freezing tolerance.