• Korean Journal of Plant Resources
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• v.32 no.3
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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 Plant Biotechnology
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• v.36 no.3
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• pp.275-280
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• 2009

Direct shoot regeneration from leaf or internode or petiole segments was conducted in 33 cultivars of chrysanthemum. Shoot regeneration rates varied according to cultivars, culture media, and explant types. The high shoot regeneration rate of more than 70% in 15 cultivars (‘Pink Pangpang’, ‘Orange Memory’, ‘Relance’, ‘Zinba’, ‘Beakma’, ‘Innocence’, ‘Sunny Pangpang’, ‘Euro Yellow’, ‘Dublin’, ‘Boramae’, ‘Peak’, ‘Euro White’, ‘Vesuvio White’, ‘Linneker Salmon’ and ‘Pink Pride’) and 2 ones (‘Forward’ and ‘Agason’) was obtained from the segments of leaves and internodes, respectively, cultured on MS medium containing 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 30 g-$L^{-1}$ sucrose. That in 6 cultivars (‘Shuhonochikara’, ‘Hakunosen’, ‘Whitney Pangpang’, ‘Plaisir D’Amour’, ‘Grace’ and ‘Kumsu’) was observed from the segments of leaves or internodes cultured on 1/2 MS medium 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 15 g-$L^{-1}$ sucrose In case of 3 cultivars (‘Ilweol’, ‘Puma White’ and ‘Sharon’), when internode explants excised from mother plants, which were pre-cultured on MS medium containing 2 g-$L^{-1}$ activated charcoal and 30 g-$L^{-1}$ sucrose for two months in the dark, and cultured on MS medium containing 1.0 mg-$L^{-1}$ BAP, 0.5 mg-$L^{-1}$ IAA and 30 g-$L^{-1}$ sucrose, that was shown. Seven cultivars including ‘Puma Yellow’, ‘Argus’, ‘Yes Morning’, ‘Whiparam’, ‘Hakunohikari’, ‘Charming Eye’ and ‘Moon light’ requires more improved culture conditions. Tissues with the highest shoot regeneration rate were in descending order, leaf, petiole, and internode segments.

• Korean Journal of Medicinal Crop Science
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• v.15 no.2
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• pp.73-81
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• 2007

An efficient regeneration system was developed using leaf, petiole, and internode explants. Highly embryogenic callus was obtained following cultivation on MS basal nutrient supplemented with 2 $mg/{\ell}$ 2,4-D. Globular, heart, torpedo and cotyledon shaped somatic embryo were produced from the surface of embryogenic callus. Direct shoot regeneration without intermediate callus formation has been achieved on MS medium supplemented NAA and BAP. The percentage of response varies with different concentration of auxin and cytokinin treated individually or in combination. The best shoot regeneration response (54.28%) and number of shoot per explant (12.67) were achieved on the medium supplemented with 0.1 $mg/{\ell}$ NAA and 1 $mg/{\ell}$ BAP. The regenerated shoot transformed into young plant when cultured into elongation and root induction medium. More than 90% of in vitro propagated plants could survive when transferred to the greenhouse for acclimation. This optimized regeneration system can be used for rapid shoot proliferation and genetic transformation.

• Plant Biotechnology Reports
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• v.5 no.2
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• pp.187-195
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• 2011

The gesneriaceous perennial plant Titanotrichum oldhamii has beautiful foliage and attractive bright yellow flowers. However, breeding of T. oldhamii by conventional sexual hybridization may be difficult because sexual reproduction of this species is very rare. In the present study, plant regeneration systems via both direct and indirect formation of adventitious shoots from leaf explants were established as the first step toward breeding T. oldhamii by using biotechnological techniques. Adventitious shoots were formed efficiently on medium containing $0.1mg\;l^{-1}$ benzyladenine. Histological observation showed that shoot formation on this medium occurred directly from leaf epidermal cells without callus formation. On the other hand, leaf explants formed calluses on medium containing $0.1mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid. The calluses could be maintained by monthly subculturing to fresh medium of the same composition. When the calluses were transferred to plant growth regulator-free medium, they formed adventitious shoots. Directly and indirectly formed shoots rooted well on medium containing $0.1mg\;l^{-1}$ indole-3-butyric acid. Plantlets thus obtained were successfully acclimatized and grew vigorously in the greenhouse. Flow cytometry analysis indicated that no variation in the ploidy level was observed in plants regenerated via direct shoot formation, whereas chromosome doubling occurred in several plants regenerated via indirect shoot formation. Regenerated plants with the same ploidy level as the mother plants showed almost the same phenotype as the mother plants, whereas chromosome-doubled plants showed apparent morphological alterations: they had small and crispate flowers, and round and deep green leaves.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.175-182
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• 2009

Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments from mature plus trees on MS liquid medium supplemented with $0.25mg\;1^{-1}$ KIN and $0.25mg\;1^{-1}$ IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment, while $0.25mg\;1^{-1}$ KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength MS medium with $0.15mg\;1^{-1}$ IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing 1:1 mixture of vermicompost and soil at $27{\pm}2^{\circ}C$ for hardening and acclimatization. 14- to 15-week-old well-established hardened plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation, providing a value addition to Eastern Cottonwood propagation in future.

• Journal of Plant Biotechnology
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• v.2 no.1
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• pp.51-54
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• 2000

Leaf explants of the cucumber (Cucumis sativus L.) were cultured on Murashige and Skoog's (MS) medium supplemented with various concentrations of $\alpha$-naphthalene acetic acid (NAA) and 6-benzylaminopurine (BAP). Direct shoot orgnogenesis as well as callus formation with somatic embryos and multiple shoots was observed from leaf explants of cvs. Shinhukjinju and Chungjang. The highest frequency of shoot formation 80% was observed on MS medium supplemented with NAA/BAP (5.0 ${\mu}{\textrm}{m}$/2.5 ${\mu}{\textrm}{m}$), with explants forming 3-7 shoots. Shoots formation occured within 3 to 4 weeks. Only one subculture of calli was required for plant regeneration on normal growth regulator-free medium. Plantlets transferred to soil developed into plants of normal appearance, which flowered and set fruits.

• Plant Biotechnology Reports
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• v.2 no.1
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• pp.7-11
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• 2008

A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings as well as mature field-grown plants cultured on Murashige and Skoog's (MS) medium supplemented with thidiazuron (TDZ) ($2.27{\mu}M$), 6-benzylaminopurine (BA) ($2.22{\mu}M$) and indole-3-butyric acid (IBA) ($0.49{\mu}M$). The presence of TDZ in the induction medium has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented with BA ($4.44{\mu}M$), kinetin (Kn) ($2.33{\mu}M$), indole-3-acetic acid (IAA) ($1.43{\mu}M$), and gibberellic acid ($GA_3$) ($0.72{\mu}M$). Well-developed shoots were rooted on MS medium supplemented with IBA ($0.5{\mu}M$) after 30 days. Regenerated plants after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation.

• Korean Journal of Weed Science
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• v.8 no.2
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• pp.182-186
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• 1988

This study was conducted to level up the tolerance of tobacco plant against bialaphos herbicide through tissue culture. The relatively good shoot regeneration from the subcultured calli treated with bialaphos at 0.5 ppm was observed in old the tobacco varieties tested such as NC 82, BY 4 and KA 101. However, at the treatment of bialaphos 1.0 ppm, shoot regeneration was only made in KA 101 variety, showing better regeneration than that of untreated one, When these shoots were transfered to the medium containing of bialaphos 10.0 ppm, the percentage of living shoots (i.e. tolerant plant) was very low, showing 2.43% in NC 82, 2.76% in KA 101 and 0.78% BY 4. Calli were induced and multiplied from leaf petiole of the above tolerant plants even under 2.5ppm of bialaphos, showing an average of 9% in NC 82 and 16% in KA 101 as compared with the untreated control. No calli were induced from tolerant plants as bialaphos concentration increased up to 5.0 ppm. Direct shooting from leaves of the above tolerant plants, that is selected at 10.0ppm of bialaphos treatment, was observed even under 10.0ppm of bialaphos treatment both in NC 82 and in KA 101 varieties, indicating that tolerance of tobacco plants against bialaphos can be greatly increased.

• Journal of Plant Biotechnology
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• v.49 no.4
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• pp.325-330
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• 2022

In this study, a plant tissue culture system was established for Atractylodes spp., an economically valuable medicinal crop in Korea that has low domestic production and is increasingly imported. In particular, Atractylodes ovata was treated with four types of cytokinins, 6-benzylaminopurine (BA), zeatin, kinetin, and thidiazuron (TDZ), in two different concentrations (0.5 and 1.0 mg/L). Among the four types of cytokinins, the BA treatment was effective for the shoot and root growth of A. ovata. Both the 0.5 mg/L and 1.0 mg/L concentrations of BA showed similar results; however, the 1.0 mg/L concentration of BA was more effective in promoting shoot and root growth. The treatments showed that the TDZ treatment was not effective for the shoot and root growth, except for the number of shoots and the fresh weight (FW) of the root; therefore, it was unsuitable for this species. In this study, we established a mass production system of A. ovata. Our results showed that direct in vitro regeneration may make a significant contribution to improving the cultivation of the medicinal plant A. ovata.

• Journal of Plant Biotechnology
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• v.34 no.2
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• pp.153-159
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• 2007

Plant regeneration of Pulsatilla koreana was achieved via adventitious shoot formation indirectly from callus and directly from adventitious root segments. For the callus induction from leaf or petiole explants, combination of 2,4- dichlorophenoxyaceticacid (2,4-D) with $2.22\;{\mu}M$ 6-benzyladenine (BA) was effective. Adventitious shoot induction from callus was enhanced by the combined treatment with $0.1\;{\mu}M$ polyvinylpyrrolidone (PVP) compared to cytokinin treatment alone. Adventitious roots were induced from the petiole segments on 1/2 MS medium with $4.93\;{\mu}M$ IBA. High frequency direct adventitious shoot formation from the segments of adventitious roots was achieved on medium with $4.92\;{\mu}M$ 2-isopentenyladenine (2-ip). Elongated shoots were rooted on half-strength MS medium containing $5.71\;{\mu}M$ indole acetic acid (IAA). Regenerated plantlets with well-developed shoots and roots were successfully transferred to soil. This in vitro propagation protocol might be useful for mass propagation as well as conservation of this plant.