• Journal of Plant Biotechnology
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• v.35 no.3
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• pp.209-213
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• 2008

This study was conducted to regenerate shoots from internode sections(about 1mm in thickness) of Ardicia pusilla de Candolle. Internode sections were cultured on MS medium supplemented with TDZ or both TDZ and IBA. At one month after culture, primodium, which looks like protocorm like body(PLB) of orchid, appeared around swollen internodes. And then it grew and changed into the shape similar to granule of orange at two or three months after culture. At four to five months after culture, explants covered with them became a cluster, and then multiple shoots were regenerated from them. Primodia formation was the best when internode was cultured on MS medium supplemented with 0.25 $mg{\cdot}L^{-1}$ thidiazuron(TDZ) and 0.5 $mg{\cdot}L^{-1}$ indole-3-butyric acid(IBA). That internodes were cultured on MS medium supplemented with either higher concentration of TDZ than that of IBA, or equal concentration of TDZ and IBA, or TDZ only was little effective for primodia formation.

• Current Research on Agriculture and Life Sciences
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• v.15
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• pp.47-52
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• 1997

The experiment was carried out to find the optimal condition for seed germination in vitro of Calanthe species. Seeds of Calanthe spp. after 90 days self pollination were well germinated in the MS medium. Germination condition of cross seeds between C. discolor and C. striata was much improved by pretreatment of micro sonic waves for 10 min. The days of germination was more shortened. and protocorm growth was more promoted in microsonic wave treatment than in non-treatment.

• Journal of Plant Biology
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• v.22 no.1_2
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• pp.35-43
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• 1979

The present investigation has been made to study the deficiency symptoms of boron on the formation of cell wall and the development of the individual components of the orchid cell wall. Analytical samples were taken from two sources; one from the individual orchid plants started from an apical meristem culture followed by the generation of the protocorm-like body which was developed into a plant, the other from the plant cultivated in water for 30 days. The amount of boron in the cultrues were controlled and the deficiency symptoms were observed under theelectron microscope, optical microscope with samples taken from the zones of elongation of leaves and compared the dry weight of cell walls and finally the various fractions of the cell wall components. The following results were obtained: (1) The growth of roots and leaves was hampered in the boron deficient plants. (2) In the boron-deficient leaves a severe necrosis and cracks were developed in the tissue of zone of elongation besides the decrease in growth. (3) under the electorn microscope the cell walls of boron-deficient plants showed rough undulated structures unlike the smooth control cell walls. (4) the dry weight of total cells and cell walls of boron deficient plants were higher than the control plants. (5) In the boron deficient plant the amout of pectin and hemicellulose isolated from cell walls were higher and the amount of protein was lower than the controlled plots.

• Plant Biotechnology Reports
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• v.4 no.2
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• pp.157-163
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• 2010

A protocol for in vitro propagation from pseudobulb sections of Lycaste armomatica (Graham ex Hook) Lindl., an ornamental and fragrant orchid, was developed. The effect of four cytokinins: kinetin (K), metatopolin (mT), $N^6$-benzyladenine (BA), and thidiazuron (TDZ), in equimolar concentrations, was investigated. Shoot formation from apical and basal pseudobulb sections was obtained in all treatments. A few medial sections cultured in media supplemented with BA formed protocorm-like bodies. Shoot formation was greater from the basal section than the apical, and mainly occurred in explants cultured in media containing TDZ. The highest average numbers of shoots per explant were achieved from basal sections cultured in media supplemented with TDZ at 4.4, 8.87 and 2.2 ${\mu}M$, forming on average 9.9, 8.6 and 7.3 shoots per explant, respectively. Since the medial pseudobulb section was the worst explant for propagation of L. aromatica, we recommend that pseudobulbs be divided into two sections; the basal half should be cultured in MS medium supplemented with TDZ at 4.4 ${\mu}M$ and the apical half with TDZ at 2.2 ${\mu}M$. Subculturing individual shoots in MS medium without plant growth regulators allows further development and rooting. A survival rate of more than 90% under greenhouse conditions was achieved. This research represents a direct contribution to the conservation and sustainable use of this valuable natural resource.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.145-152
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• 2008

Aerides vandarum and Vanda stangeana are two rare and endangered vandaceous orchids with immense floricultural traits. The intergeneric hybrids were synthesized by performing reciprocal crosses between them. In vitro germination response of the immature hybrid embryos was found to be best on half-strength Murashige and Skoog medium supplemented with 20% (v/v) coconut water/liquid endosperm from tender coconut. Determination of hybridity was made as early as the immature seeds or embryos germinated in vitro, using randomly amplified polymorphic DNA (RAPD) markers. Out of 15 arbitrarily chosen decamer RAPD primers, two were found to be useful in amplification of polymorphic bands specific to the parental species and their presence in the reciprocal crosses. However, a decisive profile that can identify the reciprocal crosses could not be provided by RAPD. Amplification of the trnL-F non-coding regions of chloroplast DNA of the parent species and hybrids aided easy identification of the reciprocal crosses from the fact that maternal inheritance of chloroplast DNA held true for these intergeneric hybrids. Subsequent restriction digestion of the polymerase chain reaction (PCR) amplified trnL-F non-coding regions of chloroplast DNA also consolidated the finding. Such PCR-based molecular markers could be used for early determination of hybridity and easy identification of the reciprocal crosses.

• Korean Journal of Plant Resources
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• v.35 no.4
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• pp.508-514
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• 2022

Bulbophyllum auricomum Lindl. is very popular among orchid growers due to the attractive fragrance of its flowers and has become an endangered orchid in Myanmar. In this study, we carried out an aseptic technique that can be used to conserve this endangered orchid species. The seeds of B. auricomum Lindl. were obtained from artificial pollination and cultured in MS basal medium for seed germination. The effect of coconut water and BAP in MS basal medium on callus induction was investigated. The highest callus induction was found at 2.0 mg/L BAP. The maximum growth of protocorm-like bodies (PLBs) was evaluated, and the best response was observed on MS medium supplemented with 150 mL/L coconut water at pH 5.6. MS basal medium supplemented with 150 mL/L coconut water along with 2.0 mg/L BAP and 1.0 mg/L NAA (MCBN) showed the highest number of plantlets at 15℃ at the second week of culture. At the second and third week of culture, MS medium supplemented with 2.0 mg/L BAP and 1.0 mg/L NAA (MBN) showed the best result in terms of the number of leaves and the longest leaves at 15℃ and 25℃, respectively. The present study showed evidence of successful in vitro propagation of B. auricomum Lindl.

• 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.

• Horticultural Science & Technology
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• v.33 no.6
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• pp.900-910
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• 2015

This study aimed to investigate the efficiency of colchicine and oryzalin in inducing polyploidy in two Cymbidium hybrids [Showgirl 'Silky' and Mystery Island 'Silk Road' (Silk Road-4)]. Colchicine was used at concentrations ranging from 50 to $500mg{\cdot}L^{-1}$, with treatments lasting 1 to 3 weeks. Oryzalin was used at concentrations ranging from 3 to $20mg{\cdot}L^{-1}$, with treatments lasting 3 to 6 days or 1 to 3 weeks. The survival rate of PLBs was better in colchicine than in oryzalin solutions. The ploidy levels were screened using flow cytometry. In C. Showgirl 'Silky', the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (60%) and the 2-week treatment in $5mg{\cdot}L^{-1}$ oryzalin (46.7%). In C. Mystery Island 'Silk Road' (Silk Road-4), the highest chromosome doubling efficiencies were obtained with the 1-week treatment in $50mg{\cdot}L^{-1}$ colchicine (16.7%) and the 3-day treatment in $10mg{\cdot}L^{-1}$ oryzalin (6.7%). Colchicine was more efficient than oryzalin in terms of polyploidy induction. Furthermore, pre-treatment, which entailed poking 10 times with forceps, improved the efficiency of chromosome doubling.

• 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.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.163-168
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• 2012

An optimization in vitro seed germination was established by using triphenol tetrazolium chloride (TTC) test, which has been known as two rare orchids (Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama) in Jeju island. We have established proper NaOCl treatment for in vitro germination of G. verrucosa and H. sikokiana through TTC test. In the case of H. sikokiana, seed viability through TTC test was high with 95% in control. However, NaOCl 1% treatment for 30 minutes showed the highest embryo swelling rate to seed viability. Likewise, swelling formation of embryos, diameter of embryos, protocorm formation and diameter of protocorms of G. verrucosa was 87%, $59{\mu}m$, 91% and $138{\mu}m$ through NaOCl 1% treatment for 30 minutes. This result will be applied on the basic information for improving in vitro seed germination rate of G. verrucosa and H. sikokiana.