Abstract
Main conclusion
Endogenous auxin determines the pattern of adventitious shoot formation. Auxin produced in the dominant shoot is transported to the internodal segment and suppresses growth of other shoots.
Abstract
Adventitious shoot formation is required for the propagation of economically important crops and for the regeneration of transgenic plants. In most plant species, phytohormones are added to culture medium to induce adventitious shoots. In ipecac (Carapichea ipecacuanha (Brot.) L. Andersson), however, adventitious shoots can be formed without phytohormone treatment. Thus, ipecac culture allows us to investigate the effects of endogenous phytohormones during adventitious shoot formation. In phytohormone-free culture, adventitious shoots were formed on the apical region of the internodal segments, and a high concentration of IAA was detected in the basal region. To explore the relationship between endogenous auxin and adventitious shoot formation, we evaluated the effects of auxin transport inhibitors, auxin antagonists, and auxin biosynthesis inhibitors on adventitious shoot formation in ipecac. Auxin antagonists and biosynthesis inhibitors strongly suppressed adventitious shoot formation, which was restored by exogenously applied auxin. Auxin biosynthesis and transport inhibitors significantly decreased the IAA level in the basal region and shifted the positions of adventitious shoot formation from the apical region to the middle region of the segments. These data indicate that auxin determines the positions of the shoots formed on internodal segments of ipecac. Only one of the shoots formed grew vigorously; this phenomenon is similar to apical dominance. When the largest shoot was cut off, other shoots started to grow. Naphthalene-1-acetic acid treatment of the cut surface suppressed shoot growth, indicating that auxin produced in the dominant shoot is transported to the internodal segment and suppresses growth of other shoots.
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Abbreviations
- 4-Cl-PEO-IAA:
-
4-Chloro-α-(phenyl ethyl-2-one)-indole-3-acetic acid
- Kyn:
-
L-Kynurenine
- NAA:
-
Naphthalene-1-acetic acid
- NPA:
-
N-(1-Naphthyl)phthalamic acid
- PIN:
-
PIN-FORMED
- PPBo:
-
4-Phenoxybenzeneboronic acid
- TIBA:
-
2,3,5-Triiodobenzoic acid
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Acknowledgements
This study was supported by the Inoue Enryo Memorial Foundation for Promoting Science from Toyo University (KI), and by the Research Center for Life and Environmental Sciences, Toyo University. We thank Shosaku Kashiwada, Hiroki Higashibata (Toyo University), and Uma Maheswari Rajagopalan (Shibaura Institute of Technology) for their constructive comments on this study.
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425_2020_3367_MOESM1_ESM.tiff
Supplementary file1 (TIFF 9526 kb) Fig. S1 Time course of adventitious shoot formation after NAA treatment. Data are means ± SE (n = 3)
425_2020_3367_MOESM2_ESM.tiff
Supplementary file2 (TIFF 9526 kb) Fig. S2 Effect of TIBA on adventitious shoot formation on internodal segments after 0–8 weeks of culture. Data are means ± SE (n = 3). n.f., adventitious shoot formation was not found. Different letters above bars indicate significant difference (Tukey’s HSD, P < 0.05)
425_2020_3367_MOESM3_ESM.tiff
Supplementary file3 (TIFF 9526 kb) Fig. S3 Effect of NPA on adventitious shoot formation on internodal segments after 0–8 weeks of culture. Data are means ± SE (n = 3). n.f., adventitious shoot formation was not found. Different letters above bars indicate significant difference (Tukey’s HSD, P < 0.05)
425_2020_3367_MOESM4_ESM.tiff
Supplementary file4 (TIFF 9526 kb) Fig. S4 Patterns of adventitious shoot formation on internodal segments placed vertically on phytohormone-free culture medium. a Internodal segments with adventitious shoots formed. Bar, 5 mm. b Numbers of adventitious shoots formed after 6 weeks of culture. c Internodal segments were partitioned into four regions, and adventitious shoots formed were counted in each region. Data are means ± SE (n = 3). n.s., not significant (t-test, P ≥ 0.05)
425_2020_3367_MOESM5_ESM.tiff
Supplementary file5 (TIFF 9526 kb) Fig. S5 IAA levels in internodal segments and adventitious shoots after 10 weeks of culture. a Apical region of the internodal segment. b Basal region of the internodal segment. c Lower part of the shoot stem. d Upper part of the shoot stem. e Leaf. Red lines indicate cut sites. Data are means ± SE (n = 4). Different letters above bars indicate significant difference (Tukey’s HSD, P < 0.05)
425_2020_3367_MOESM6_ESM.tiff
Supplementary file6 (TIFF 9526 kb) Fig. S6 Effects of auxin concentration on adventitious shoot formation. Bar, 5 mm. Adventitious shoots (white circles) were normally formed on the apical side of an internodal segment. In contrast, a small number of adventitious shoots were formed on the basal side at a lower concentration of auxin. High concentration of auxin induced adventitious roots on callus formed on the basal side. Optimum concentration of auxin is required for adventitious shoot formation
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Koike, I., Watanabe, S., Okazaki, K. et al. Endogenous auxin determines the pattern of adventitious shoot formation on internodal segments of ipecac. Planta 251, 73 (2020). https://doi.org/10.1007/s00425-020-03367-5
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DOI: https://doi.org/10.1007/s00425-020-03367-5