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Propagation of Hinoki Cypress ( Chamaecyparis obtusa ) Through Tissue Culture Technique as a Sustainable Method for Mass Cloning of Selected Trees

Author

Listed:
  • Tsuyoshi E. Maruyama

    (Department of Forest Molecular Genetics and Biotechnology, Forestry and Forest Products Research Institute, Matsunosato 1, Tsukuba 305-8687, Japan)

  • Momi Tsuruta

    (Department of Forest Molecular Genetics and Biotechnology, Forestry and Forest Products Research Institute, Matsunosato 1, Tsukuba 305-8687, Japan)

  • Asako Matsumoto

    (Tama Forest Science Garden, Forestry and Forest Products Research Institute, Todori 1833-81, Hachioji 193-0843, Japan)

  • Ryouichi Kusano

    (Kumamoto Prefectural Forestry Research and Instruction Center, 8-222-2, Kurokami, Kumamoto 860-0862, Japan)

  • Tetsuji Hakamata

    (Shizuoka Prefectural Research Institute of Agriculture and Forestry, 2542-8, Negata, Hamamatsu 434-0016, Japan)

Abstract

Propagation of hinoki cypress (Japanese cypress, Chamaecyparis obtusa , Cupressaceae) through adventitious bud multiplication was performed using leaf-segment explants from cutting plants of selected adult trees. Explants were successfully surface-sterilized (>90% asepsis) by agitating them in 2.5% (w/v available chlorine) sodium hypochlorite solution for 15 min and then rinsed with sterile distilled water. Explants approximately 2 cm long were cultured on plates containing medium supplemented with 6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D), 20 g/L sucrose, and 7 g/L agar. The cultures were kept at 25 ± 1 °C under a 16-h photoperiod with a photon flux density of approximately 65 µmol m −2 s −1 . The optimal adventitious bud multiplication (31.5 buds per explant) was obtained on a medium supplemented with 10 µM BAP in combination with 1 µM 2,4-D. Proliferated adventitious buds were elongated better on medium supplemented with 1 µM trans -zeatin. The best rooting result (86%) was achieved on a rooting medium supplemented with 1 µM 3-indolebutyric acid in combination with 0.1 µM 1-naphthaleneacetic acid. However, rooting response varied according to genotypes. Clones related to the cultivar ‘Nangouhi’ (Na18, Na14 x Isa, Na14-14, Isa x Na14, and NaS) were easier to root than those derived from the cultivar ‘ShizuokaKenZairai’ (SKZ5 and SKZ8). Regenerated plantlets did not show morphological abnormalities and showed a high survival rate after acclimatization (>90%).

Suggested Citation

  • Tsuyoshi E. Maruyama & Momi Tsuruta & Asako Matsumoto & Ryouichi Kusano & Tetsuji Hakamata, 2025. "Propagation of Hinoki Cypress ( Chamaecyparis obtusa ) Through Tissue Culture Technique as a Sustainable Method for Mass Cloning of Selected Trees," Sustainability, MDPI, vol. 17(7), pages 1-14, March.
    • Handle: RePEc:gam:jsusta:v:17:y:2025:i:7:p:3039-:d:1623435
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    References listed on IDEAS

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    1. Bates, Douglas & Mächler, Martin & Bolker, Ben & Walker, Steve, 2015. "Fitting Linear Mixed-Effects Models Using lme4," Journal of Statistical Software, Foundation for Open Access Statistics, vol. 67(i01).
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