Tissue culture literation
| Plant latin name | Cinnamomum cassia Blume | |||||||||||||||||||||||||||||||||||||||||||||
| Original code | Cinnamomum_cassia-Exp-1 | |||||||||||||||||||||||||||||||||||||||||||||
| Publication | 吉松ら．AROMA RESEARCH, 21 (3), 211-219 (2020) | |||||||||||||||||||||||||||||||||||||||||||||
| Objectives | Development of an efficient in vitro propagation system for Cinnamomum cassia Blume | |||||||||||||||||||||||||||||||||||||||||||||
| Materials | Cinnamomum cassia seeds obtained from the greenhouse-cultivated plants at Takeda Garden for Medicinal Plant Conservation, Kyoto, Takeda Pharmaceutical Company Limited in 2017 (9 seeds, weight of 1 seed: 210.8±33.1 mg) | |||||||||||||||||||||||||||||||||||||||||||||
| Explant | Seeds | |||||||||||||||||||||||||||||||||||||||||||||
| Explant sterilization | Seeds were surface sterilized by immersion in a 75% ethanol solution for 1 min, rinsed with sterile Milli-Q water and then in sterilization solution (3% sodium hypochlorite with 0.1% v/v Tween- 20) for 30 min, rinsed seven times with sterile Milli-Q water. | |||||||||||||||||||||||||||||||||||||||||||||
| Initial culture | Nine sterilized seeds were placed on a half macro salts Murashige and Skoog medium containing 2% sucrose [(2)/2 MS]and solidified with 0.25% w/v gelrite (30 ml/3 cm i.d. ×15 cm culture tube, 1 seed/tube) and cultured at 20℃ in the dark. Rooted seeds were transferred onto Woody Plant（WP） mediurm containing 3% sucrose, 0.25% gelrite, glutamine 10 mg/L (WPG HF) at 23℃ under 14 hr/day light (LED, Pale Pink, NKsystem at 8,000 Lux). All the seeds germinated without expanding the cotyledons until the day 238 after sowing. Germination frequency at day 131 was 88.9%. | |||||||||||||||||||||||||||||||||||||||||||||
| Shoot multiplication | The axenic seedlings were subjected to the micropropagation studies. Multiple shoots formed in the presence of 3-indolebutyric acid (IBA) 1 mg/L and 6-benzyladenine (BA) 3 mg/L from the apical shoot segment of the seedling, the subsequent cultured shoot and the plantlet. | |||||||||||||||||||||||||||||||||||||||||||||
| Rooting | The shoot segment prepared from the multiple shoot rooted in the presence of IBA 1 mg/L at a ratio of 37%. Theoretically, 22 plantlets could be obtained from a seed using this micropropagation system within one and a half year. | |||||||||||||||||||||||||||||||||||||||||||||
| Acclimation | In vitro rooted plantlet was washed with a weak stream of tap water, transplanted in a pot filled with vermiculite and kept in a greenhouse (over 20℃). The aerial part of the plantlet was coverred with a rap to maintain high humidity for a week. The pots were set on the tray which was filled with oxidized glutathion solution (1 mg/L) or water for irrigation for the first 2weeks from transplant. After 2 weeks, the plantlet were irrigated with water. In vitro plantlet obtained by culturing shoot segment with IB 0.5 mg/L were successfully acclimatized with water supply and acclimatized plants could be potted for further growth. This results imply GSSG is not necessary for the acclimatization. | |||||||||||||||||||||||||||||||||||||||||||||
| Planting | The acclimatized plants were potted in the earthen bowl (15 cm i.d.) filled with soil-compost-nursery soil for vegetable and flower (Takii & Co., Ltd)-sand (6:2:1:1) and cultivated in a greenhouse (over 20℃). Plants grew healthily. | |||||||||||||||||||||||||||||||||||||||||||||
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| Traints of regenerant | ||||||||||||||||||||||||||||||||||||||||||||||
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