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

Tsuyoshi E. Maruyama (), Momi Tsuruta, Asako Matsumoto, Ryouichi Kusano and Tetsuji Hakamata
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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

Sustainability, 2025, vol. 17, issue 7, 1-14

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%).

Keywords: Cupressaceae; clonal propagation; bud organogenesis; multiple shoots; sustainable forestry (search for similar items in EconPapers)
JEL-codes: O13 Q Q0 Q2 Q3 Q5 Q56 (search for similar items in EconPapers)
Date: 2025
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