• Korean Journal of Plant Tissue Culture
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• v.25 no.1
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• pp.1-5
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• 1998

This study describes the effect of the growth regulators such as 2,L-D and BA, on the structural abnormalities of somatic embryos derived from leaf explants of Angelica gigas Nakai, Also, the relationship between the cotyledon number of a somatic embryo and its germinability is explored. Embryogenic calli were selected from calli formed on explants cultured on MS solid basal medium supplemented with 0.5mg/L 2,4-D, 1mg/L 2,4-D, 1mg/L plus 0.1mg/L BA, and 1 mg/L 2,4-D plus 0.5mg/L BA. Cotyledonary abnormalities were observed in somatic embryos which were developed from embryogenic calli cultured on MS medium containing 1mg/L 2,4-D for 8 weeks and then subcultured on 2,4-D free MS medium for 3 weeks. The frequency of abnormalities was as follows: 22.8% one cotyledon, 42.5% two cotyledons, 16.8% three cotyledons, 7.8% four cotyledons, 1.8% five cotyledons, and 8.2% jar shaped cotyledon. In addition, ABA treatment indicated an improvement of the somatic embryo with normal cotyledon (65.3%). ABA was important role to the high production of normal somatic embryos. Two cotyledon embryos showed germinability 77.8%. However the germinability of somatic embryos with anomalous cotyledons was prominently low: One cotyledon, 62.5%; three cotyledons, 43.3%; four cotyledons, 60%; five cotyledons, 50% and jar shaped cotyledon, zero%. Thus, germinability was essentially, inversely proportional to cotyledon number.

• Journal of Plant Biotechnology
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• v.33 no.1
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• pp.27-32
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• 2006

To clarify the roles of bromelain in plants, we isolated BL1 gene encoding bromelain from pineapple stem tissues and sequenced. The full length cDNA is 933 bp and encodes a polypeptide of 311 amino acid residues. The cDNA is most similar 94% at the amino acid level to bromelain previously isolated from pineapple (BAA21929). Explants of Lactuca sativa were co-cultivated with Agrobacterium tume-faciences LBA 4404 strains containing nptII and BL1 gene for transformation. Through initial selection of regenerated explants by culturing on a kanamycin and carbenicillin containing MS medium, multiple shoots were obtained after 2 months of culture. For a complementary step of selection, putative transgenic shoots were transferred to 1/2 Ms basal medium supplemented with 100 mg/L kanamycin and 500 mg/L carbenicillin. The selected shoots were obtained T1 generation seeds with emasculation, and tested with PCR analysis using 35S promoter and BL1 specific primers whether BL1 gene was introduced to genome of the plants. These results confirmed that produced the specific PCR bands in the putative transgenic lines. Additionally the Northern blot and endo protease activity showed that transcripts of BL1 gene were detected in transgenic lines. Theses results suggest that BL1 gene be successfully integrated and transcripted in the transgenic lettuce plants.

• Journal of Plant Biotechnology
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• v.33 no.1
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• pp.49-56
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• 2006

Gerber (Gerbera hybrida) is a valuable ornamental species grown as a potted plant and cut flowers. However, genetic variability within the gerbera genus is very limited. So it is absolutely needed to introduce and widen genetic resources into gerbera lines by genetic transformation. For the purpose, 18 Korean gerbera lines were screened to establish Agrobacterium-mediated genetic transformation procedure. In an experiment to select Korean gerbera lines which are amenable to Agrobacterium-inoculation, 12 lines turned out to be positive in Agrobacterium-inoculation. More callus were produced from BA 2ppm, Zeatin 2ppm, IAA 0.2ppm in pre-culture and regeneration medium (2X media) but there was no difference in the frequency of GUS expression rate. In another experiment to find out optimal condition for highly efficient Agrobacterium-inoculation, petiole and leaf explants have been treated with four different pre-culture periods, two different co-culture periods and two different Agrobacterium strains. As a result, high GUS expression has been showed from petiole and leaf explants treated no pre-culture period with LBA4404 Agrobacterium tumerfaciens, 5 day co-culture period and dipping treatment.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.37-41
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• 1997

The in vitro regeneration and genetic transformation systems in hot pepper(Capsicum annuum L.) have not been routinely available, which has been a major limiting factor in the application of new genetic manipulations. An efficient procedure to regenerate whole pepper plants and to generate transgenic plants expressing a foreign gene was established. A relatively high frequency of plant regeneration was observed when hypocotyl and cotyledon explants were cultured on MS medium supplemented with NAA 0.1 mg/L plus zeatin 2.0 mg/L or IBA 10.0 mg/L plus BAP 1.0 mg/L. Addition of AgNO$_3$5 $\mu$M to these media improved the regeneration frequency up to 8%. For plant transformation, hypocotyl and cotyledon explants of hot pepper were precultured on shoot induction media without kanamycin added for 2 days, and then cocultured with Agrobacterium tumefaciens pDY183 for 2 days. Putative transformants were obtained from selection media containing 100 mg/L kanamycin sulfate and 500 mg/L carbenicillin. Putatively selected transformants were confirmed by amplification of selectable marker genes (ADA and NPT II) by polymerase chain reacion. Successful transcripts of ADA gene were detected by Northern blot analysis. Enzyme activity of ADA was also examined by spectrophotometric analysis, and expression of ADA gene in hot pepper suggests the potential application of ADA gene as a selectable marker in plants.

• Journal of Bio-Environment Control
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• v.11 no.3
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• pp.115-120
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• 2002

Growth and development of Limonium spp.‘Ocean Blue’plantlets were studied under three levels of photosynthetic photon flux (PPF),70,150 and 220 $\mu$mol. $m^{-2}$ . $s^{-1}$ , two levels of $CO_2$ concentration, 500 and 1000 $\mu$mol. $m^{-1}$ , and two levels of number of air exchanges per hour (NAEH),0.1 $h^{-1}$ and 2.8 $h^{-l}$. Explants were obtained from photomixotrophically-micropropagated plantlets. Four explants per vessel were cultured under cool-white fluorescent lamps for 16 h. $d^{-1}$ at 25$\pm$11$^{\circ}C$ and 70~80% relative humidity. In treatments of 2.8 $h^{-1}$ NAEH, a 10 mm round hole made on the vessel cap was sealed with a microporous filter and two $CO_2$ concentrations in the culture rooms were provided from a liquefied $CO_2$ tank. Fresh and dry weights, height, length of the longest root, number of loaves, and leaf area significantly increased with increasing PPF and especially, $CO_2$ concentration. Growth was enhanced by a 2.8 $h^{-1}$ NAEH. Overall, treatment with a 220 $\mu$mol. $m^{-2}$ . $s^{-1}$ PPF and a 1000 $\mu$mol. $m^{-1}$ $CO_2$ resulted in the most vigorous growth of Limonium spp. ‘Ocean Blue’ plantlets.s.

• Journal of Plant Biotechnology
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• v.35 no.2
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• pp.155-161
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• 2008

This paper reported the establishment of mass production system of adventitious roots of Codonopsis lanceolata through shake flask and bioreactor culture. Induction of adventitious roots was started from the explants of leaf, stem and root on 1/2 strength Murashing and Skoog (MS) solid medium. Stem segments were the best explants for induction of adventitious roots compared to leaf and root segments. Among the different auxins tested (NAA, IBA and IAA), number of adventitious root per explant was highest on solid medium with 1.0 mg/L IBA, and produced $9.9{\pm}1.2$ roots per explant. However, growth of adventitious roots was fast in the presence of IBA at low concentration (0.1 mg/L). In shake flask culture, maximum production of adventitious roots (fresh weight) was obtained in half-strength MS medium compared to full-strength and one-third MS medium. When the adventitious roots produced in shake-flask culture were transferred to 5 L air-lift bioreactor, 16 times of fresh weight increase was gained after one month of culture. These results indicate that this protocol for the production of C. lanceolata adventitious roots can be applied to large scale culture for practical application.

• Journal of The Korean Society of Grassland and Forage Science
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• v.23 no.1
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• pp.59-64
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• 2003

Comparisons of callus formation ratios from seed explants, callus sizes, regeneration ratios from callus and regeneration efficiency for 4 orchardgrass (Dactylis glomerata L.) varieties (Three were developed in Korea and one was imported from foreign nation) are as follows; Jangbeol 102 (67.0%) has the highest callus formation ratio in 4 weeks incubated callus after bedding the seed explants, but Potomac (68.4%) has the highest ratio in 6-week callus. Potomac (3.91cm) has the highest callus size in 4-week callus, but Jangbeol 101 (4.32cm) has the highest size in 6-week callus. Jangbeol 101 (17.7%) has the highest plant regeneration ratio in 4-week callus, but Potomac (37.4%) has the highest ratio in f-week callus. Jangbeol 102 (11.5%) has the highest plant regeneration efficiency in 4-week callus, but Potomac (25.6%) has the highest efficiency in 6-week callus.

• Korean Journal of Plant Resources
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• v.24 no.5
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• pp.483-488
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• 2011

Cacalia firma recently has been used increasingly as leaf vegetables but endangered in natural forest. In this work, we established the plant regeneration via adventitious shoot formation from petiole segments of seedling and in vitro plantlets. Wounding of seed coats and $GA_3$ treatments were effective to induce in vitro germination of seeds, whereas, seed did not germinate at all without these treatment. When cotyledon, leaf, petiole, and root segments of seedling were cultured on medium with 2 $mg{\cdot}L^{-1}$ benzyl adenine (BA) and 0.5 $mg{\cdot}L^{-1}$ naphthaleneacetic acid (NAA), petiole segments showed highest number of shoots per explant among the other segments. Among the various kinds of cytokinins, BA, isopentyl adenine (2-ip), kinetin, zeatin, thidiazuron (TDZ), TDZ and BA treatments were effective to induce high frequency of adventitious shoot formation from petiole segments of in vitro propagated plants. NAA stimulated the frequency of adventitious shoot formation but not for number of adventitious shoots per explants compared to TDZ or BA treatment alone. Most of adventitious shoots were developed directly from surfaces of explants. Adventitious shoots were transferred on medium with IBA for root formation, thereafter the plantlets were successfully transferred to soil.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.80-80
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• 2018

Hosta is a genus of the family Asparagaceae and distributed in East Asia. There are six Hosta species (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) native to Korea and among them, four species (H. minor, H. jonesii, H. venusta and H. yingeri) are endemic to the Korea peninsula. Hosta is generally propagated by seed, crown division or tissue culture. However, tissue culture is a more efficient method to mass proliferation, a new cultivar development and disease-free plantlet production in a limit time. Hence, we conducted this study to evaluate the influence of various plant growth regulators (PGRs) treatments on the induction of callus and somatic embryo of the six Hosta species. Leaf, petiole and root were used to select optimum tissue culture explants. Petiole explants thus only were used for callus induction and somatic embryogenesis with TDZ (0.1, 0.5 or 1.0mg/L) and NAA (0.1 or 0.5 mg/L) combinations. After 12 weeks of culture, the highest rate of somatic embryogenesis was achieved on modificated MS medium containing 1.0 mg/L TDZ and 0.1 mg/L NAA in H. capitata and H. minor (15.5%, respectively), 0.1 or 0.5 mg/L TDZ and 0.1 mg/L NAA in H. jonesii (22.2%), 1.0 mg/L TDZ and 0.5 mg/L NAA in H. yingeri (26.7%), and 0.1 mg/L TDZ and 0.5 mg/L NAA in H. venusta (53.3%). H. clausa showed very low effect on somatic embryogenesis by PGRs; 2.2%. There was interspecies difference to PGRs respond for callus and somatic embryo induction. Regenerated multiple shoots and plantlet of H. minor, H. jonesii, H. venusta and H. yingeri were obtained via somatic embryogenesis.

• Korean Journal of Medicinal Crop Science
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• v.2 no.2
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• pp.154-161
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• 1994

This experiment was carried out to establish micropropagation system in Fritillaria thunbergii Miq. Through the culture of bulblet scales, stems, node-buds and shoot tips with special reference to the effect of physiological age of explant and plant growth regulators on bulblet formation. Number of formed bulblets was significantly increased in node-bud or stem tissue compared to scals segments and on the medium supplemented with kinetin than BA containing medium. Optimum levels of kinetin for bulblet formation from node-bud taken from above 3 cm shoot length and stem segments excised from below 3 cm shoot length were 5.0 mg /L and $1.0{\sim}3.0\;mg$ /L kinetin, respectively. Interesting phenomenon was observed, the direct formation of bulblets from the axilliary bud of cultured explants. Bulblet forming capacity in stem tissue was depended on stem age, young stem had high regeneration ability compared to old stem taken from above 10 cm shoot length. 1.0 mg /L kinetin was optimum concentration for the formation of bulblets from old stem segments. Stem tissue taken from underground growing plant was promoted coampare to shoot tips or bulb scale segments. Optimum concentration of sucrose was $5{\sim}7%$. Summariged above results revealed that effective explant for micropropagation was stem and /or node-bud tissue excised from less than 3 cm plant height compared to those of bulb scale segments which showed high contamination after culture. Maximum multiplication rate of young stem and /or node-bud segment was about 20 times. Kinetin requirement for stimulation of bulblet formation from cultured explant depended on source of explants but favorable levels of kinetin for organogenesis ranged from 1.0 mg /L to 5.0 mg /L.