• Journal of The Korean Society of Grassland and Forage Science
• /
• v.19 no.3
• /
• pp.273-280
• /
• 1999

Hypocotyl explants of Medicago saliva cv. Vernal were cultured on Murashige and Skoog (MS) medium supplemented with various combinations of growth regulators. After six weeks of culture, somatic embryos were formed from calli on MS medium containing $4mg/{\ell}$ 2,4-D and $0.1mg/{\ell}$ kinetin, or $4mg/{\ell}$ 2,4-D and $0.5mg/{\ell}$ kinetin. The mature somatic embryos were developed to plantlets when subcultured on MS basal medium. In order to obtain secondary somatic embryogenic calli, cotyledon of regenerated plantlets were cultured on a callus induction medium. Embryogenic calli were formed on MS medium containing $4mg/{\ell}$ 2,4-D alone. For induction and development of secondary somatic embryogenesis, the embryogenic calli were transferred to MS basal medium containing either 2,4-D or NAA. Multi-secondary somatic embryogenesis was the most effective on MS basal medium with $0.1mg/{\ell}$ 2,4-D. The rate of secondary somatic embryo formation of regenerated plants was 18 times higher than that of seed grown plants. The mature secondary somatic embryo were germinated into plantlets on MS basal medium after six weeks of culture.

• Korean Journal of Plant Resources
• /
• v.24 no.4
• /
• pp.461-471
• /
• 2011

This study was carried out to establish an in vitro culture method of callus having a high antioxidant activity from Ginkgo biloba L. Leaf explants were cultured on Murashige and Skoog's medium supplemented with various growth regulators. The explants were incubated in the dark or 3,000 lux cool-white light. Methanol extracts from incubated callus were evaluated for scavenging activity of the free radicals using DPPH. The best callus growth rate was achieved in MS medium combined with 10 ${\mu}M$ NAA and 5 ${\mu}M$ kinetin in the light condition. Total antioxidant activity of cell aggregates in suspension culture [MS medium supplemented with 10 ${\mu}M$ NAA in the light] was up to 80% of ascorbic acid. By means of HPLC analysis, quantification of the quercetin dehydrate and keamperol profiles from suspension callus was compared. Contents of quercetin dehydrate and keamperol from leaf extracts were 0.07 and 2.24 ${\mu}g/20{\mu}l$, and those from callus 0.56 and 0.18 ${\mu}g/20{\mu}l$, respectively.

• Journal of Animal Science and Technology
• /
• v.46 no.5
• /
• pp.879-886
• /
• 2004

Effects of culture mediwn supplements and osmotic stress treatment on embryogenic callus induction and somatic embryogenesis were investigated in order to optimize tissue culture conditions of alfalfa(Medicago sativa L.). SH mediwn containing 5mgIL 2,4-D and 0.2mgIL kinetin was optimal for embryogenic callus induction from cotyledon tissue of alfalfa. Somatic embryos were formed when the embryogenic callus was cultured on SH mediwn supplemented with ImgIL 2,4-D and 2mgIL BA. Supplementation of 5mM L-proline and IgIL casein hydrolysate into the regeneration mediwn further increased plant regeneration frequency. Osmotic stress treatment of callus appeared to improve the frequency of somatic embryo formation, but the frequency of somatic embryo formation differed by the osmotic stress treatment using different osmotic stressors. The highest plant regeneration frequency of 30.7% was observed when embryogenic callus was treated with 0.7M sucrose for 18h. Efficient regeneration system established in this study will be useful for molecular breeding of alfalfa through genetic transformation.

• Applied Biological Chemistry
• /
• v.43 no.3
• /
• pp.184-189
• /
• 2000

Callus was induced from the petioles of scented-geranium (Pelargonium graveolense) in MS medium containing various concentrations of plant growth regulators. The highest frequency of more than 70% of callus was induced in 2 mg/l NAA and 0.5 mg/l BAP or 2 mg/l 2,4-D and 0.5 mg/l BAP combined treatment, while not in 2,4-D, NAA or BAP alone. When the callus was transferred to the MS medium containing 0.05 mg/l NAA and 0.5 mg/l BAP, were highest intensity of shoot formation, 14 shoots/callus, was induced after 5 weeks. The highest rooting was observed on hormone-free rooting media from the regenerated shoots after 3 weeks, indicating that the regeneration from geranium callus might be possible by changing the hormone ratios. Peroxidase (POD) specific activities in callus induced from 2 mg/l NAA and 0.5 mg/l BAP were higher than those of 2 mg/l 2,4-D and 0.5 mg/l BAP callus during the entire culture periods. POD isozyme C3 was the main cathodic POD isozyme expressed in NAA and BAP callus, while C1 was the main in 2,4-D and BAP callus. However, anodic POD isozymes, A1, A2 and A3 were expressed with similar activities in both hormone combinations.

• Korean Journal of Plant Tissue Culture
• /
• v.25 no.3
• /
• pp.195-200
• /
• 1998

Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

• Proceedings of the Korean Society of Grassland Science Conference
• /
• 2001.06a
• /
• pp.84.1-84
• /
• 2001

• Journal of Animal Science and Technology
• /
• v.47 no.6
• /
• pp.1067-1074
• /
• 2005

In order to optimize tissue culture conditions of Kentucky bluegrass(Poa pratensis L.), effects of culture medium supplements, media and cultivars on embryogenic callus induction and regeneration of plants were investigated. MS medium containing 3mg/L 2,4-D and 0.1mg/L BA was optimal for embryogenic callus induction from mature seeds. The highest plant regeneration frequency(57.7%) was observed when the embryogenic calli were cultured on N6 medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Among several basic media, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. Genotype was an important factor in plant regenerability. ‘Newport’ showed to have higher regeneration frequency of 53.4%. Regenerated plants were grown normally when shoots transplanted to the soil. A short tissue culture period and high-frequency regeneration system would be beneficial for molecular breeding of Kentucky bluegrass through genetic transformation.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.19 no.1
• /
• pp.23-30
• /
• 1999

The conditions for callus formation and plant regeneration were confirmed in four varieties of alfalfa(Medicago sativa L.). Among four varieties of alfalfa, "Vernal" expressed the highest rate for both of callus formation and plant regeneration. Otherwise, among SH(Schenk and Hildebrandt), MS(Murachige and Skoog) and N6 medium (Chu), SH medium was highest degree of efficiencies respectively in callus formatio and plant regeneration. In this study, we determined volume of hormones and other compounds appended in media. For callus formation, only $3mg/{\ell}$ of 2,4-D (2,4-dichlorophenoxy acetic acid) was appended in their media. For plant regeneration, the three kinds of media were used; the medium appended $5mg/{\ell}$ of NAA (1-naphtalene acetic acid) and $2mg/{\ell}$ of kinetin (6-furfurylaminopurine), the medium appended $11mg/{\ell}$ of 2,4-D and $1mg/{\ell}$ of kinetin, and the medium appended $1.6g/{\ell}$ of ammonium sulfate and $5.75g/{\ell}$ of proline. We obtained alfalfa plants from callus by regeneration, about sixty five days later transfer calli to regeneration media.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.1
• /
• pp.35-39
• /
• 1995

In order to investigate the effects of gelling agent on rice anther culture, anthers of rice (Japonica cv Daecheongbyeo) were cultured on N$_{6}$ media supplemented with 0.8, 1.2 or 1.6% Junsei agar and 05, 0.4, 0.6, 0.8 or 1.0% Gelrite (Phytagel, Sigma). On Junsei agar media, the frequency of callus induction was decreased in proportion to agar concentration. The frequency of callus induction was more increased as 67.6% and 54.8% in media containing 0.4 and 0.6% Gelrite than in agar media. The frequency of plant regeneration and spontaneous doubled-diploid was directly proportional to Junsei agar and Gelrite concentration. The number of green and spontaneous doubled diploid plant was highest on 0.6% Gelrite medium. In order to optimize the concentration of growth regulators for the callus induction medium containing 0.6% Gelrite, anthers were cultured on N$_{6}$ media supplemented with 2mg/L NAA, 2 mg/L 2,4-D, 1mg/L NAA and 1mg/L 2, 4-D, or 1mg/L NAA, 1mg/L 2,4-D and 0.5mg/L kinetin. The maximum frequency of callus induction and plant regeneration was obtained from the medium supplemented with 2 mg/L NAA and 0.6% Gelrite. In conclusion the induction of embryogenic callus, the frequency of plant regeneration and in vivo chromosome doubling was more effective in Gelrite media than in Junsei agar media.dia.

• Current Research on Agriculture and Life Sciences
• /
• v.10
• /
• pp.1-9
• /
• 1992

Culture condition for callus formation and plant regeneration were optimized by the selection of explants and the manipulation of hormonal combination in the culture medium. The calli induced from seed, cotyledon, hypocotyl and mesophyll segments were more vigorously proliferated under dark condition than those under continuous light condition. Hypocotyl-and cotyledon-derived calli were more regenerative as compared with those of seed and mesophyll. Callus formation from hypocotyl and cotyledonary explants was enhanced on MS medium with 1.0 mg/${\ell}$ 2, 4-D and 0.1 to 0.5 mg/${\ell}$ kinetin or BAP. The combination of 0.1 mg/${\ell}$ NAA and 2.0 to 4.0 mg/${\ell}$ kinetin was the most effective for shoot regeneration from the callus. The maximum frequency (24.0%) of shoot regeneration was obtained from the hypocotyl-derived callus transferred to MS medium supplemented with 0.1mg/${\ell}$ NAA and 4.0 mg/${\ell}$ kinetin. The capacities for callus, root and shoot formation from cotyledon and hypocotyl explants were remarkably different among cultivars of B. napus tested. The calli induced from hypocotyl produced more shoots than those from cotyledon.