Abstract
Graptopetalum paraguayense E. Walther is an edible succulent plant that is native to Mexico. When leaves of G. paraguayense are detached from the mother plant, shoots and roots regenerate from the leaf base. To investigate the endogenous hormonal regulation of plant regeneration in G. paraguayense, we evaluated the effects of applying gibberellin (GA), abscisic acid (ABA), and phytohormone biosynthesis inhibitors to leaves detached from the mother plant. GA3 and paclobutrazol did not affect plant regeneration of G. paraguayense, whereas ABA treatment significantly suppressed plant regeneration. When leaves were treated with the ABA biosynthesis inhibitor fluridone at 10 μM, the frequency of shoot regeneration was more than twice that of the control on the 6th day after leaf detachment. In addition, when both fluridone and ABA were applied at the same time, the frequency of plant regeneration decreased to control levels or below. Endogenous ABA in leaves was approximately 1.3 ng gFW−1 just after leaf detachment, decreased to 0.3 ng gFW−1 by the 1st day after leaf detachment, and increased to 3.0 ng gFW−1 by the 9th day after leaf detachment. These findings indicate that ABA plays an important role in suppression of plant regeneration in G. paraguayense. Although plant regeneration during storage and transportation currently reduces the economic value of G. paraguayense, our findings may provide a method to delay unwanted plant formation at the leaf base.
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Acknowledgements
We thank Mr. Akira Sugimoto (Agri Assist Japan Corporation) and Mr. Masayuki Chiba (Kashiwa Senko Co., Ltd.) for providing the young plantlets of G. paraguayense. We also thank Dr. Shosaku Kashiwada and Dr. Hiroki Higashibata (Toyo University) for their constructive comments. This study was supported by the Inoue Enryo Memorial Foundation for Promoting Science from Toyo University to T.T., and was in part supported by the Research Center for Life and Environmental Sciences, Toyo University.
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Figure S1
Inner tissue structure at the base of a detached leaf. (a) A leaf base of G. paraguayense. Bar = 3 mm. (b) Observation of the leaf base (indicated by arrow in (a)). The red dotted line in (b) indicates the plane of sectioning. Bar = 1 mm. (C–G) Frozen sections of the leaf base stained with toluidine blue O. In all sections, the leaf base is toward the left side of the figure. (c) 0 day, (d) 2 days, (e) 4 days, (f) 6 days, and (g) 8 days after leaf detachment. Yellow arrows show cell layer with active cell division. Black arrow shows a regenerating shoot. Bars = 100 μm. Figure S2 Effects of GA3 (a) and paclobutrazol (b) on plant regeneration from leaves of G. paraguayense. Photographs show regenerated shoots and roots on the 18th day after leaf detachment. Bars = 1 mm. Figure S3 Effects of ABA (a) and fluridone (b) on plant regeneration from leaves of G. paraguayense. Photographs show regenerated shoots and roots on the 18th day after leaf detachment. Bars = 1 mm. Figure S4 Effects of combining fluridone and ABA on plant regeneration from leaves of G. paraguayense. Both ABA and fluridone were applied to the basal end of detached leaves every 3 days. (a) Regenerated shoots and roots on the 18th day after leaf detachment. Bar = 1 mm. (b) Frequency of shoot regeneration from the detachment site. (c) Frequency of root regeneration from the detachment site. Values are means ± SEM (n = 3). +, #, *, and × indicate significant differences between the indicated treatment and the control (0 μM fluridone) (Student’s t-test, P < 0.05). In the figure key, “A” indicates 10 μM ABA and concentrations indicate the level of fluridone. Data of the 9th day shown in Supplemental Figs. S4b and S4c are picked up in Figs. 5a and 5b, respectively. Figure S5 Effect of water supplementation during plant regeneration in G. paraguayense. (A) Leaves cultured with or without 50 ml of Milli-Q water in a plastic plate (105 × 195 × 25 mm). (B) Leaves cultured on dry or wet Kimwipe (moistened with 10 ml Milli-Q water) in a plastic plate (105 × 195 × 25 mm). Values are means ± SEM (n = 3). Student’s t-test, *P < 0.05, n.f.: not found, n.s.: not significant. Bars = 3 cm. (C) Regeneration from a detached leaf on the 18th day after leaf detachment with or without water supply. Bars = 1 mm. Figure S6 Comparison of ABA levels in each region of a detached leaf (0–12 days after detachment) and the stem of the mother plant. Bar = 1 cm. Values are means ± SEM (n = 4). In the graphs for 1 d and 6 d, values marked with the same letter were not significantly different (Tukey’s HSD, P < 0.05). S, stem; n.d., not detected; n.s., not significant. Figure 6 shows the same data analyzed by leaf region. (PDF 9703 kb)
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Tamaki, T., Kubo, S., Shimomura, K. et al. Effects of Gibberellin and Abscisic Acid on Asexual Reproduction from Graptopetalum paraguayense Leaves. J Plant Growth Regul 39, 1373–1380 (2020). https://doi.org/10.1007/s00344-020-10068-6
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DOI: https://doi.org/10.1007/s00344-020-10068-6