• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.375-380
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• 2003

This study was conducted to establish a practical masspropagation system of Phalaenopsis clones from basal shoot segments. The frequency of PLB (protocorm like body) induction was compared with various explants. Basal shoot segments showed the most successful result of 45％, while root tips, stalk node segments, stalk leaves and mature leaves represented low frequency (below 5％). The PLB induction ratio in the culture of basal shoot segments was examined with 11 different Phalaenopsis varieties, and the majority of varieties, including pink flower lines, showed an about 30％ rate of PLB formation. Especially, when whole basal shoot parts without cutting were inoculated onto PLB induction medium, giant PLB was induced from explant. This giant PLB was green color and big in size compared with normal PLB. When dissected giant PLB segments inoculated onto PLB multiplication medium, only normal size of PLBs were induced from them. PLBs induced by basal shoot culture were transferred onto proliferation medium and then shooting medium, from which normal plants were formed. Therefore, this culture method is considered as effective and practical protocol for Phalaenopsis mericlone production. In addition, it is suggested that clones of an infinite number can be produced consecutively by this culture system through repeated cycles of PLB induction and proliferation using the basal shoot segment of flask plant.

• Korean Journal of Plant Resources
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• v.31 no.4
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• pp.372-377
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• 2018

This study was investigated the effect of immature rhizome production according to harvest times-based treatment method for seed production of Gastrodia elata. The results revealed that when the tuber weight of G. elata harvested in spring (GEHS) was ${\geq}100g$, the rate of artificial fertilization, protocorm formation, and immature rhizome formation was 90.9%-94.8%, 3.1%-5.4%, and 10.1%-15.3%, respectively. When G. elata harvested in fall (GEHF) was treated at a low temperature for 4 weeks or more, the rate of artificial fertilization, protocorm formation, and immature rhizome formation was 70.4%-87.6%, 2.2%-2.6%, 8.7%-9.5%, respectively. Therefore, to produce seeds and immature rhizomes, GEHS must have tubers of more than 100 g, whereas GEHF requires breaking dormancy by low-temperature treatment for 4 weeks or more. Compared with those of GEHS, the rate of artificial fertilization, protocorm formation, and immature rhizome formation was lower in GEHF; however, it was higher than those in the natural germination state. Thus, it can be expected that G. elata can be produced throughout the year by ensuring that the seeds and immature rhizomes of G. elata are produced using a constant tuber weight and by breaking dormancy with low temperature treatment.

• Horticultural Science & Technology
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• v.16 no.1
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• pp.42-44
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• 1998

The effect of medium composition on PLB formation and multiplication and shoot regeneration was studied to establish the micropropagation system of the tropical orchid Phalaenopsis. The highest frequence of PLB formation resulted from the VW medium with 1.2 times ion concentration, 1% sucrose, 1.5g/L PVP or 2.5g/L active charcoal, apple and potato extract and 4g/L gellan gum. The highest ratio of PLB multiplication was obtained from the VW medium with 2% sucrose, apple and potato extract and cotton plate. The shoot regeneration was the most effective with the hyponex medium with 3% apple, 3% potato and 4% banana extract.

• Korean Journal of Plant Resources
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• v.21 no.4
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• pp.341-345
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• 2008

Two orchid species of Calanthe discolor Lindl. and C. discolor for. sieboldii (Decne.) Ohwi, which have different form flower color and size. They were crossed in mid April by artificial pollination, and the F1 hybrid seeds were collected mid October. Germination of seeds was investigated on pre-treatment of seeds and under the various environmental conditions. Germination was promoted by moisture absorption and ultrasonic treatment of seeds. Dark culture of F1 hybrid seeds enhanced germination and protocorm formation, and development into seedlings compared with light culture. Although, plant growth regulators such as NAA and BA had a slightly promotive effect on seed germination and protocorm growth, regenerated seeding were showed abnormal growth patterns. Regenerated F1 hybrid plantlets were successfully transferred to pot.

• Horticultural Science & Technology
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• v.16 no.2
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• pp.236-238
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• 1998

This study was conducted to investigate the effective media and optimal days after pollination for germination of seeds which were crossed by selfing or reciprocal pollination in Cymbidium spp. The elongation of ovary stopped about 110 days after pollination regardless of cross combinations. The increase of ovary width stopped about 140 days after pollination showing some differences among cross combinations. The cross combinations expressing high capsule number and viability showed good germination when they were sown in vitro. The capsules collected 150 to 180 days after pollination showed good germination performance and short days of germination. Hyponex media was the most effective for protocorm development and germination.

• Journal of Plant Biotechnology
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• v.38 no.4
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• pp.293-300
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• 2011

This study is conducted to develop an efficient transformation system via particle bombardment with PLBs (Protocorm-like bodies) in Cymbidium. For this, pCAMBIA3301 vector which carries a herbicide-resistant bar gene and gus gene as a reporter gene was used for transformation with Cymbidium cultivars 'Youngflower ${\times}$ masako' line. To select transformants, proper concentration of herbicide, PPT (phosphinotricin), should be determined. As a result, 5 mg/l of PPT was selected as a proper concentration. Further, proper conditions for particle bombardment were determined to obtain a high frequency of transformation. Results showed that 1.0 ${\mu}g$ of DNA concentration, 1,100 and 1,350 psi for helium gas pressure, 1.0 ${\mu}m$ of gold particle and 6 cm of target distance showed the best result for the particle bombardment experiment. Also, pre-treatment with combination 0.2 M sorbitol and 0.2 M mannitol for 4 hrs prior to genetic transformation increased the transformation efficiency up to 2.5 times. Using transformation system developed in this study, 3.2 ~ 4.0 transgenic cymbidium plants can be produced from 100 bombarded PLBs on average. Putative transgenic plants produced in this system confirmed the presence of the bar gene by PCR analysis. Also, leaves from randomely selected five transgenic lines were applied for Basta solution (0.5% v/v) to check the resistance to the PPT herbicide. As a result, three of them showed resistance and one of them showed the strongest resistance with the maintenance of green color as non-transformed plants showed. Using this established transformation system, more genes of interests can be introduced into Cymbidium plants by genetic transformation in the future.

• Journal of Bio-Environment Control
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• v.31 no.2
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• pp.133-141
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• 2022

Bulbophyllum auricomum Lindl. is a rare orchid and has flowers with an attractive fragrance. The present study investigated the tissue culture method for micropropagation. Capsules derived from artificial self-pollination were obtained for the best seed germination in MS basal medium. Plant growth regulators (1.0 mg·L^-1 of BAP and 2.0 mg·L^-1 of NAA) were affected by callus induction from subcultured pseudobulb explants. For the callus subculture, different natural plant extracts were tested in 11 treatment media. Among them, MS medium with 150 mL·L^-1 of coconut water was generally effective in fresh weight (1.75 ± 0.08) and (3.01 ± 0.20) of callus proliferation and PLBs induction at 1 and 2 months, respectively, followed by an MS combination of 30 g·L^-1 of banana and 20 g·L^-1 of potato extract. The results of a comparative study of different MS mediums containing plant growth regulators with a natural extract combination and MS medium supplemented with natural extract only showed that MS medium supplemented with a combination of natural extracts (150 mL·L^-1 of coconut water) and plant growth regulators (2.0 mg·L^-1 of BAP and 1.0 mg·L^-1 of NAA) obtained the highest shoot regeneration (3.37 ± 0.17) and (6.41 ± 0.68) after 1 month and 2 months of culturing, respectively.