• Journal of Plant Biotechnology
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• v.40 no.4
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• pp.203-209
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• 2013

Genetic manipulation of pear (Pyrus pyrifolia Nakai) breeding is still difficult due to lack of reliable regeneration system. The aim of this research is to establish shoot regeneration system from leaf explants for pear (P. pyrifolia cv. Niitaka) using various concentrations of plant growth regulators and carbon source supplemented to medium. The highest regeneration rate of about 20% was found on a medium containing 4.4 g/L of Murashige and Skoog (MS) without vitamins, Linsmaier and Skoog (LS) vitamins were added separately. Leaf explants of pear were cultured on MS medium containing 7 g/L of Daishin agar supplemented with various concentrations of NAA (0.01, 0.05, 0.1, 0.5 mg/L) in combination with BA(3, 5, 10 mg/L) for shoot regeneration. In medium with 5 mg/L of BA and 0.01 mg/L of NAA, adventitious shoot regeneration rate was higher than others treated. The optimal results were observed using MS medium supplemented with 30 g/L sorbitol as carbon source on regeneration system. Sorbitol is considered better carbon source than sucrose for shoot regeneration of pear (P. pyrifolia cv. Niitaka). In order to increase of shoot regeneration in pear (P. pyrifolia cv. Niitaka), plant agar and Daishin agar used as gelling agents, Daishin agar is more efficient in shoot regeneration.

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.187-193
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• 2005

In an effort to optimize tissue culture conditions for genetic transformation of tall fescue (Festuca arundinacea Schreb.), an efficient plant regeneration system from seed-derived calli was established. MS medium containing 6 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.1 mg/L benzyladenine (BA) were optimal for embryogenic callus formation from mature seed and had a strong effect on successive plant regeneration. The plant regeneration frequency above 50% was observed when embryogenic calli induced in this medium were transferred to N6 medium supplemented with 1 mg/L 2,4-D and 3 mg/L BA. Among several basic media, MS and N6 medium were optimal for callus induction and plant regeneration, respectively. 'Kentucky-31' showed to have high frequencies of embryogenic callus induction and plant regeneration up to 58.3 and 50%, respectively. Addition of sucrose to the regeneration medium as a carbon source increased regeneration frequency up to 55%. A short tissue culture period and high-frequency regeneration system established in this study will be useful for molecular breeding of tall fescue through genetic transformation.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.514-518
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• 1992

The present study has been perromed to investigate the optimal conditions for protoplast formation and regeneration of oleandomycin-producing Streptomyces antibioticus (S. antibioticus) KCTC 1081. Mycelia were grown in YME medium containing 0.2% (w/v) glycine and converted into the protoplast by incubating at 35.deg.C for 60 minutes in protoplast buffer (P buffer) containing 4 mg/ml lysozyme. The reversion of protoplasts to the normal filamentous state was examined by the growth on various synthetic agar media. A high reversion rate was obtained by incubating the protoplasts on a hypertonic agar medium containing 20 mM $Mg^{++}$, 5 mM $Ca^{++}$ and 0.3 M sucrose at 28.deg.C for 5 days. From these experiments, we established the improved regeneration medium and a protocol which supports higher and more consistent levels of regeneration of S. antibioticus protoplasts. The regenerant showed an increased antimicrobial activity compared with that of the initial strain.n.n.

• Journal of Plant Biotechnology
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• v.6 no.4
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• pp.253-258
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• 2004

Plant regeneration depending on explant type was inves-tigated with cotyledon, hypocotyl, and leaf explants of garland chrysanthemum (Chrysanthemum coronarium L.) cultured on MS basal medium supplemented with various concentrations of SAP and NAA combination. Among the three different types of explants, hypocotyl explants grown on MS medium containing $1.0{\mu}M\;NAA,\;1.0{\mu}M\;BA\;and\;6{\mu}M\;AgNO_3$ produced the highest adventitious shoots (4.67 per explant). Hypocotyl explants not only produced more vigorous shoots, which regenerated aster than the cotyledon and leaf explants. An efficient root formation was observed in MS medium containing $3\%$sucrose. The concentration of NAA did not show significant effects on root formation. Results from this experiment suggested that hypocotyl explants were efficient for the regeneration of garland chrysanthemum.

• Archives of Pharmacal Research
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• v.16 no.4
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• pp.300-304
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• 1993

Genetic transformation of streptomyces gaespitosus by plasmid plJ 702 was camied out. Optimal conditions for the protoplast preparation of streptomyces casepitosus, its regeneration, and its transformation by plJ 702 were evaluated. Addition of 2% glycine to the culture broth was optimal for protoplast yield. Formation and regeneration of protoplasts were most efficient when the mycelium were harvested at between late log and stationary growth phase. The regeneration frequency of the protoplasts was 15% when the protoplats were regenerated on R2YE agar media containing 0.5M sucrose. Under the best condition for protoplats (M.W. 4,000) treatment for 2 minutes.

• Korean Journal of Medicinal Crop Science
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• v.7 no.4
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• pp.275-281
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• 1999

Embryogenic calli of Codonopsis lanceolata were cultured on MS agar medium containing various concentrations of sucrose as a carbon source. Upon transfer to MS basal medium, somatic embryos of cotyledonary stage converted to plantlets. When sucrose was added with greater than 4%, the number of shoots and roots regenerated from somatic embryo increased. However, the growth of shoots and roots was retarded in agar medium with more than 2% sucrose, but promoted in medium with lower concentration of sucrose. Saponin contents of shoots regenerated from somatic embryos, embryogenic calli, non-embryogenic calli, and native roots were determined by HPLC. Saponin contents of native root was variable, depending on regenerant, embryogenic calli, and cotyledonary embryos. The saponin contents of regenerated roots in medium with high sucrose was similar to native roots. Saponins content based on cell differentiation to shoot and root was dramatically decreased. This results could be effectively controlled for the production of useful secondary metabolites.

• Korean Journal of Plant Tissue Culture
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• v.27 no.2
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• pp.101-107
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• 2000

The explants of yacon (Polymnia sonchifolia Poeppig & Endlicher) were cultured to invest th8e dedifferentiation condition, and formative callus from leaf was cultured to find the regeneration and micro-crown bud formation. Basal MS medium was more effective to form callus than 1/2 MS and B$_{5}$ medium. Calli formations from leaf, petiole and lateral bud were more effective on MS medium supplemented with 1.0, 2.0 mg/L 2,4-D and 0.2, 0.4 mg/L kinetin or BA than 1.0, 2.0 mg/L NAA and 0.2, 0.4 mg/L kinetin or BA. Formative callus from leaf was proliferated about 70% on medium supplemented with 1.0 mg/L BA. When callus was proliferated, 63% regeneration rate was shown on medium supplemented with 1.0, 2.0 mg/L BA in case of subculture for 3~4 months but was not shown on medium supplemented with 1.0, 2.0 mg/L kinetin. Micro-crown bud formed as addition of BA at 3~4 months after callus culture and then was obtained many at 5~6 months, it was most formed about 82% on medium supplemented with 5 mg/L BA. Rate of micro-crown bud formation was increased as more over 5 mg/L BA concentration, when this time, however, shoot had thick leaves and short internodes, and then withered before long, Micro-crown bud was formed about 88.0% on medium supplemented with 5% sucrose, that was more increased 28% than with 3% sucrose. The buds of crown bud between harvested in field and formed in vitro were difference only in size, but both were similar in shape according to histological view.

• Korean Journal of Plant Resources
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• v.18 no.1
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• pp.154-160
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• 2005

To establish rapid micropropagation through organogenesis from apices-derived callus or direct adventitious shoot of three calla lily cultivars(Zantedeschia spp, cv. Sunlight, cv. Chiante, cv. Pink Persuation) were cultured on Murashige and Skoog medium supplemented with different plant growth regulators. The formation rate of callus, organogenesis and in viかo tuber production among the three cultivars were tested. Callus was obtained from cvs. Sunlight, Chiante and Pink Persuasion; the best cultivar was Sunlight. Sunlight induced $53.3\%$ callus and Chiante had the highest rate of $56.7\%$ direct shoot regeneration on medium with 2.0 mg/L BA. Regeneration frequencies ranged from 20 to $70\%$ on medium with 2.0-3.0 mg/L BA. The highest percentage of regeneration and the greatest number of shoots were obtained on medium containing 3.0 mg/L BA in three cultivars. Cytokinins induced multiple shoot formation; 1.0 mg/L of 2ip, 5.0 mg/L of BA, and 1.0 m/L of BA induced 16, 14 and 12 multiple shoots in cvs. Sunlight, Chiante and Pink Persuasion, respectivly. 1.0 mg/L of IAA enhanced root growth in cvs. Sunlight and Chiante while cv. Pink Persuasion exhibited enhanced root growth at 2.0 mg/L of IBA. NAA, however, induced no change in root growth. The addition of 90 g/L sucrose enhanced in vitro tuber formation and following tuber expansion in cv. Sunlight, while 70 g/L of sucrose was effective in cvs. Chiante and Pink Persuasion.

• Plant Biotechnology Reports
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• v.4 no.4
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• pp.261-267
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• 2010

We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg $1^{-1}$ ${\alpha}$-naphthaleneacetic acid (NAA) and 0.1 mg $1^{-1}$ 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing 1.0 mg $1^{-1}$ BA and 1.0 mg $1^{-1}$ NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg $1^{-1}$ $GA_3$ had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained by 1% sucrose. Most secondary embryos (87.2-94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose.

• Korean Journal of Plant Resources
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• v.26 no.3
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• pp.383-388
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• 2013

The research concerned of the regeneration of plants from embryos obtained from anther cultures of ginseng (Panax ginseng C. A. Meyer). The aim was to determine the influence of the regeneration medium on the efficiency of the regeneration process. We conducted to determine the optimum conditions such as cold pretreatment, plant growth regulators and carbon sources on anther culture of P. ginseng. Highest callus formation rate was obtained when flower buds pretreated at $4^{\circ}C$ for 1 day. Among the treated growth regulators with various degrees of concentration in Murashige and Skoog's (MS) medium, 4.53 ${\mu}M$ of 2.4-dichlorophenoxyacetic acid and 4.44 ${\mu}M$ of 6-benzylaminopurine gives the most responsive callus with the frequency of 73.89% and 129.53 g of fresh weight. When we used 3-9% of sucrose and maltose among the different kinds and various concentrations of carbohydrates, callus was formed highest 67.29% in the medium with 3% of sucrose. Shoots induced from callus supplemented with 28.9 ${\mu}M$ of gibberellic acid and rooted in Gamborg's B5 medium supplemented with 14.7 ${\mu}M$ of indole-3-butyric acid.