• Current Research on Agriculture and Life Sciences
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• v.11
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• pp.11-17
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• 1993

The transfer of genetic material into pea tissue was accomplished by using an avirulent strain of Agrobacterium tumefaciens containing the binary vector. The method used for transformation requires non-tissue culture steps as it involves the inoculation of the site of the shoot removed of germinating seeds. The identification of ${\beta}$-glucuronidase activity in the tissues of $T_0$ pea plants indicates that the plant expressible ${\beta}$-glucuronidase gene, contained the T-DNA region from pLPBO2, had been transferred at least into somatic tissues. Putative transformed $T_0$ pea plants were advanced to produce $T_1$ plants which were also assayed for the presence of the transferred ${\beta}$-glucuronidase gene. The presence of the ${\beta}$-glucuronidase gene in DNAs isolated from $T_1$ plant was demonstrated by DNA gel blot hybridization. This analysis revealed that the transformed plants contained ${\beta}$-glucuronidase gene.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.53-55
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• 1997

The shoot tips of Delphinium cv. Princess Caroline were cultured on the MS medium supplemented with cytokinin and auxin alone or in combination. Among cytokinins, BA was most effective in shoot multiplication, adequqte concentrations being 1.0-5.0 mg/L. Shoot multiplication was very favorable on the media with 1.0-3.0 mg/L BA and 0.1-0.5 mg/L IAA. Additions of BA and IAA did not stimulate shoot multiplication, but increased a little fresh weight. Shoots were scarcely rooted on the media with IBA or NAA, and were not done utterly on the media containing activated charcoal. Therefore, shoots were treated by Rootone and planted in the cultural media for in vivo rooting. The highest rate of rooting was 68% in the mixed cultural medium composed of Perlite 1 and Vermiculite 1.

• Korean Journal of Plant Tissue Culture
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• v.28 no.1
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• pp.25-32
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• 2001

Shoot tips of grape were cultured in uitro and tried to identify optimal culture conditions for regeneration, multiple shoot formation from meristemoid tissue and those subsequent elongation of multi-shoots. Healthy growing shoots were taken in early May, rinsed with running tap water, soaked in a neutral detergent and washed with soft brushing, and washed out with tap water, then sterilized with 10g Ca(ClO)$_2$/140 mL distilled water (Wilson's solution) for 5 min. Survival percentage of the cultures which were sterilized as above procedures was highly increased, compared with the other sterilized method. Propagation of multi-shoots from meristemoid showed a good response in 3/4 strength MS medium enriched with 0.1 mg/L NAA and 3.0 mg/L BA. Shoot elongation from multi-shooting clump well occurred in 3/4 strength MS medium supplemented with 80 mg/L adenine sulfate, 0.1 mg/L NAA and 1.0~2.0 mg/L BA.

• Korean Journal of Plant Tissue Culture
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• v.21 no.5
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• pp.281-286
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• 1994

The hormonal regulation of organ differentiation was investigated in the tissue culture of sweet potato. Embryogenic callus was induced from shoot tips cultured on MS medium supplemented with 1 mg/L 2,4-D. When embryogenic callus was transferred to medium containing 0.1 mg/L GA$_4$, it proliferation was stimulated. The callus gave rise to plantlets when cultured on medium containing 0.1 mg/L BA. Addition of 0.1 mg/L jasmonic acid or 0.01 mg/L brassinolide to medium was effective for the development of healthy normal plantlets.

• Korean Journal of Plant Resources
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• v.31 no.6
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• pp.684-694
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• 2018

This study describes the successful establishment of a cryopreservation protocol for Citrus limon cultivars: 'Frost Eureka limon' and 'Cook Eureka limon', using a droplet-vitrification method. The shoot tips that were excised from in vitro grown seedlings of the two cultivars were preserved in liquid nitrogen (LN) and successfully regenerated into whole plants. Excised shoot tips were pre-cultured for 1 or 2 days in 0.3 M and 0.5 M sucrose solutions at $25^{\circ}C$ and incubated in a loading solution (LS) composed of 17.5% glycerol + 17.5% sucrose in Murashige and Skoog (MS) medium for 40 min at $25^{\circ}C$. Prior to direct immersion in LN for 1 h, the shoot tips were dehydrated with plant vitrification solution 2 (PVS2) at $0^{\circ}C$ or PVS3 at $25^{\circ}C$. The frozen shoot tips were re-warmed and unloaded with 1.2 M sucrose in $\text\tiny{^1/_2}$ MS for 30 min at $25^{\circ}C$. Shoot tips were post-cultured overnight on survival medium and then micrografted onto 'trifoliate orange' (Poncirus trifoliate (L.) Raf. seedling rootstocks for recovery and to produce whole plants. The highest regrowth rates were 53.5% and 50.3% for cryopreserved shoot tips of 'Frost Eureka limon' and 'Cook Eureka limon', respectively, when pre-cultured in 0.3 M and 0.5 M sucrose concentrations in a sequencing manner, with LS and treated with PVS2 for 60 min at $0^{\circ}C$. We also investigated whether the ammonium ion concentration on post-culture medium affected the viability of the cryopreserved Citrus shoot tips. The viability of cooled samples, following culturing on woody plant media (WPM) containing $\text\tiny{^1/_4}$ ammonium nitrate overnight before micrografting, was the highest (70.3%) in 'Frost Eureka limon'. The study described here is a cost-effective and safe method to conserve Citrus fruit cultivars, for the improvement and large-scale multiplication of fruit plants and for breeding disease resistance.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.46 no.6
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• pp.464-467
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• 2001

In order to establish a in vitro propagation system for gold tree[Dendropanax morbifera $L_{EV}$], the effects of auxins and cytokinins on shoot multiplication and rooting were investigated. Germination rate was the best in MS medium. The fresh weight and number of shoot were the best on the medium containing 0.1 or 1.0 mg/l BAP and 0.5 or 1.0 mg/l NAA. Shoots were successfully rooted in MS medium with 1.0 mg/l NAA. Roots were easily formed by the addition of auxins, especially 0.1 or 1.0 mg/l BAP.P.

• Journal of Plant Biotechnology
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• v.50
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• pp.82-88
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• 2023

Shoot-tip culture was used to produce clonal plants of radish stock seeds. Using 6-benzyladenine (BA), the largest number of RA2 line multi-shoots were formed with an average of 14.67 shoots on 1.33 µM BA in early seedlings and 11.33 shoots on 1.78 µM BA in juvenile seedlings. The largest number of RA4 line multi-shoots were formed with an average of 11.67 shoots on 2.22 µM BA in early seedlings and 13.67 shoots on 1.33 µM BA in juvenile seedlings. There was little difference in the significance level by BA concentration in both lines. Using Thidiazuron (TDZ), the number of RA2 line multi-shoots increased with increasing TDZ concentration, forming the largest number of multi-shoots in 0.45 µM TDZ (7.0 and 3.0 multi-shoots for early and juvenile seedlings, respectively), but few multi-shoots were formed from TDZ 2.25 and 4.5 µM. RA4 line produced almost no multi-shoots in early seedlings, and 3.7 multi-shoots were produced in 0.23 and 0.45 µM TDZ in juvenile seedlings, but not at higher concentrations. Analysis of the tissue culture seedlings grown by cultivating the generated multi-shoots with Radish Foundation seeds using SSR marker revealed a weak pattern of mutation in the generated tissue culture seedlings, but there was no mutant. In addition, in terms of root roots, both RA2 and RA4 lines generally had the best rooting, number of roots, and degree of root development in 4.9 µM indol-3- butyric acid (IBA).

• Korean Journal of Plant Resources
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• v.29 no.6
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• pp.635-641
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• 2016

Sweet potato (Ipomoea batatas L.) shoot tips grown in vitro were successfully cryopreserved by encapsulation-vitrification. Encapsulated explants are very easily manipulated, due to the relatively large size of the alginate beads, and a large number of samples can be treated simultaneously. In this study, the effects of sucrose preculture, cryoprotectant preculture, and post-warm recovery media on regrowth, following liquid nitrogen (LN) exposure, were investigated to establish an efficient encapsulation-vitrification protocol for sweet potato. Shoot tips of plants grown in vitro were precultured in 0.3 M sucrose for 2 d before encapsulation. Encapsulated shoot tips were pre-incubated in liquid MS (Murashige and Skoog) medium containing 0.5 M sucrose for 16 h, before preculturing in sucrose-enriched medium (0.7 M sucrose) for 8 h. Shoot tips were osmoprotected with 35% plant vitrification solution 3 (PVS3) for 3 h, before being dehydrated with PVS3 for 2 h at $25^{\circ}C$. The encapsulated and dehydrated shoot tips were transferred to 2 mL cryotubes, suspended in 0.5 mL PVS3, and plunged directly into liquid N. High levels of shoot formation were obtained for the cv. Yeulmi (65.7%) and Yeonwhangmi (80.3%). The regrowth rates of cryopreserved samples in Yeulmi (78.9%) and Yeonwhangmi (91.3%), following culture on ammonium-free MS medium for 5 d, were much higher than those cultured on standard MS medium (65.7% and 80.3%, respectively). This encapsulation-vitrification is a promising method for the long-term preservation of sweet potato.

• Journal of Plant Biology
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• v.37 no.3
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• pp.381-385
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• 1994

The highest degree of callus formation was obtained from the shoot tips of Dianthus superbus when cultured on the MS medium supplemented with 2.0 mg/L NAA and 0.5 mg/L BAP. Embryogenic calluses were obtained from the seperated friable calluses on MS medium containing 2.0 mg/L 2,4-D after 7-8 wk of culture. For plant regeneration, embryogenic calluses were selected and cultured on te proliferation medium. After 3 wk, somatic embryos appeared on MSK medium (0.5 mg/L NAA, 2.0 mg/L kinetin) and N6 medium (2.0 mg/L kinetin, 0.1 mg/LNAA, 0.1 mg/L 2,4-D and 2.0 g/L casein hydrolysate). When these somatic embryos were kept under continuous illumination, shoots were successfully regenerated on the both media. The shoots were rooted on MS medium supplemented with 2.0 mg/L NAA.

• Korean Journal of Plant Resources
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• v.32 no.1
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• pp.53-62
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• 2019

The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.