• Journal of Plant Biotechnology
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• 제44권3호
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• pp.335-342
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• 2017

Purple passion fruit (Passiflora edulis Sims) is one of the introduced tropical plants, an increasing interest has arisen due to its distinctive taste and attractive flavor. It is expected that passion fruit production and planted area will increase gradually in the years ahead because of high profitability and consumer's demands of healthful ingredients. So we tried to investigate the effect of plant growth regulators and antioxidants on in vitro plant regeneration and callus induction from leaf explants of passion fruit for an establishment of optimal mass propagation system. Young leaf explants of purple passion fruit were cultured in Murashige and Skoog (MS) medium containing different growth regulators and antioxidant additives to induce the shoot organogenesis. After 8 weeks, the highest embryogenic callus formation rate was obtained in MS medium supplemented with $1mg{\cdot}L^{-1}$ 6-benzylaminopurine (BAP) and $2mg{\cdot}L^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), furthermore, the shoot development via organogenesis was also observed. Silver nitrate ($AgNO_3$), which was added into the medium to minimize the adverse effects of leached phenolics, was effective for reduction of medium browning and sudden explant death. In the medium supplemented with $1mg{\cdot}L^{-1}$ BAP and $1mg{\cdot}L^{-1}$ gibberellic acid ($GA_3$), shoots were most vigorously regenerated and elongated. Most shoots rooted successfully in half strength medium with $1mg{\cdot}L^{-1}$ indol-3 acetic acid (IAA), and more than 90% of plantlets survived after 4-month acclimatization period.

• Plant Biotechnology Reports
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• 제5권2호
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• pp.147-156
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• 2011

Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.173-173
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• 2017

Phalaenopsis orchid is one of the most important flowers in flower industry. We conducted an experiment to find out the response of different concentration of chlorocholine chloride (CCC) for the in vitro regeneration of protocorm like bodies (PLBs) of Phalaenopsis 'Fmk02010'. We used five different concentrations of CCC and these were 0 (control), 0.01, 0.1, 1 and $10mgL^{-1}$ in modified MS medium and cultured for 42 days. We added two major salts ammonium nitrate ($412.5mgL^{-1}$) and potassium nitrate ($950.0mgL^{-1}$) to the MS medium for the modification. Maximum numbers of PLBs were found from media with $0.01mgL^{-1}$ of CCC (15.667) and maximum fresh weight (0.211 g) as well. The 100% PLB formation rate was also found from $0.01mgL^{-1}$ of CCC. We found 58.83% variation in number of PLBs ($R^2=0.5883$) and 47.44% variation in fresh weight ($R^2=0.4744$) to the different CCC concentrations. Our study suggested that increase in the CCC concentration negatively affect the PLBs organogenesis of Phalaenopsis. We can suggest that the addition of very low concentration of CCC in plant culture medium can increase the number, formation rate and fresh weight of PLBs of Phalaenopsis.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.177-177
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• 2017

Phalaenopsis orchid is one of the most important flowers in flower industry. We conducted an experiment to find out the response of different concentration of chlorocholine chloride (CCC) for the in vitro regeneration of protocorm like bodies (PLBs) of Phalaenopsis 'Fmk02010'. We used five different concentrations of CCC and these were 0 (control), 0.01, 0.1, 1 and $10mgL^{-1}$ in modified MS medium and cultured for 42 days. We added two major salts ammonium nitrate ($412.5mgL^{-1}$) and potassium nitrate ($950.0mgL^{-1}$) to the MS medium for the modification. Maximum numbers of PLBs were found from media with $0.01mgL^{-1}$ of CCC (15.667) and maximum fresh weight (0.211 g) as well. The 100% PLB formation rate was also found from $0.01mgL^{-1}$ of CCC. We found 58.83% variation in number of PLBs ($R^2=0.5883$) and 47.44% variation in fresh weight ($R^2=0.4744$) to the different CCC concentrations. Our study suggested that increase in the CCC concentration negatively affect the PLBs organogenesis of Phalaenopsis. We can suggest that the addition of very low concentration of CCC in plant culture medium can increase the number, formation rate and fresh weight of PLBs of Phalaenopsis.

• Journal of Plant Biotechnology
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• 제7권4호
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• pp.267-272
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• 2005

Common bean (Phaseolus vulgaris L.) plants were regenerated via organogenesis from mature embryonic axes, cultured on MS medium supplemented with ildole-3-ecetic acid (IAA) and thidiazuron (TDZ) for one week in the dark. Embryonic axillary regions were excised, longitudinally cut to split the both sides, and cultured for two weeks on MS medium supplemented with IAA and TDZ. The combination 0.5 mg $l^{-1}$ TDZ/0.5 mg $l^{-1}$ IAA presented the higher efficiency in shoot regeneration and the combination 0.5 mg $l^{-1}$ TDZ/0.25 mg $l^{-1}$ IAA presented the higher efficiency in conversion of shoots to plants. Regenerating explants were transferred to MS medium containing 1 mg $l^{-1}$ BAP for shoot development. All elongated shoots were rooted in vitro, presented normal phenotype and produced viable seeds. Histological analysis confirmed the mode of regeneration as de novo shoot organogenesis.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2002년도 춘계학술대회
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• pp.178-178
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• 2002

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.27-35
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• 2006

Plantlet regeneration in Asteracantha longifolia(L.) Nees (Acanthaceae), a medicinal herb has been achieved from seedling explants on basal MS medium. Three different seedling explants including node, internode and leaf segments on used. Of these three explant, leaf explants gave better response for both callus mediated organogenesis and direct multiple shoot induction. Number of explants showing differentiation of shout buds was higher on MS media supplemented with BA compared to kinetin. MS medium fortified with BA ($2.0mgl^{-1}$) and NAA ($0.5mgl^{-1}$) was found to be most suitable for both callus mediated organogenesis and elongation of shouts. The elongated shoots were successfully routed on MS medium fortified with NAA or IBA. Among them $0.1mgl^{-1}$ NAA or $0.2mgl^{-1}$ IBA provides better response for rhizogenesis. Regenerated plantlets were successfully established in soil where 85.4% or them developed into morphologically normal and fertile plants. RAPD profiling using four decamer primers confirmed the genetic uniformity of the regenerated plantlets and substantiated the efficacy and suitability of this protocol for in vitro propagation of A. longifolia.

• Plant Biotechnology Reports
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• 제2권3호
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• pp.213-218
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• 2008

Swertia chirata is an endangered gentian species that prefers to grow at higher altitudes. This ethnomedicinal herb is known primarily for its bitter taste caused by the presence of important phytochemicals that are directly associated with human health benefits. Due to a continuous loss of habitat and inherent problems of seed viability and seed germination, alternative strategies for propagation and conservation are urgently required to prevent the possible extinction of this species. We have formulated a reproducible protocol for the rapid propagation and conservation of this plant using leaves taken from in vitro shoot cultures. Direct induction of more than seven shoot buds per explant was achieved for the first time when the explants were placed on MS medium supplemented with $2.22{\mu}M$ N-6-benzyladenine, $11.6{\mu}M$ kinetin, and $0.5{\mu}M$ ${\alpha}-naphthalene$ acetic acid. Direct organogenesis was noted exclusively from the adaxial surface of the basal segments of leaves. Leaves closer to the apical meristem were more responsive than those farther away from the meristem. Plants raised through direct organogenesis were evaluated for their clonal fidelity by chromosomal analysis and DNA fingerprinting. Complete plants were successfully transferred to the field condition and produced viable seeds. Given the enormous potential of this age-old medicinal plant in terms of potential health-benefitting drugs, this protocol can be used for commercial propagation purposes and to initiate future genetic improvement studies.

• 한국자원식물학회지
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• 제32권3호
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• pp.237-243
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• 2019

Soybean is the one of recalcitrant legume species for shoot induction. Shoot regeneration via direct organogenesis was investigated in five soybean cultivars, 'Dawon', 'Pungsan', 'Daewon', 'Taekwang' and 'Chongdoo 1' by using cotyledonary node explants. Out of 5 soybean cultivars, an efficient shoot regeneration condition was developed in the two soybean cultivars, 'Dawon' and 'Pungsan'. When various kinds of plant growth regulators with different concentration were estimated, the optimum medium condition for shoot induction in both soybean cultivars was MS + B5 vitamin supplemented with BA at concentration 2 mg/L. In addition, shoot formation efficiency was increased with 97.09% and 93.88% by the pretreatment of BA onto the explants before in vitro culture in both cultivars. Shoot induction in 'Dawon' cultivar was originated from epidermal tissue and sub-epidermal layers when histological changes were investigated under shoot regeneration after culturing cotyledonary node segments on shoot induction medium for 0 to 21 days. Especially, cell dedifferentiation was observed from parenchyma cells to meristematic cell in 3-day cultured segments.

• Journal of Forest and Environmental Science
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• 제39권3호
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• pp.150-154
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• 2023

Heloniopsis orientalis (Liliaceae) is an important horticultural crop native to Korea. Under natural conditions, germination is poor and plant growth is delayed. Therefore, we have developed a vegetative propagation method to produce plants with vigorous growth characteristics via tissue culture. The regenerated shoots were then initiated directly from leaf explants on an MS medium containing either 0.5 to 2.0 mg/L 2,4-D or 1.0 to 3.0 mg/L BA. Healthy plantlets with adventitious roots were formed on the medium supplemented with 1.0 mg/L BA (81%). BA triggered callus initiation without caulogenesis or rhizogenesis, and callus formation was better on the half-strength MS medium than on the full-strength medium. This in vitro propagation protocol will be useful for conservation, as well as for mass propagation.