• Journal of Ginseng Research
• /
• v.38 no.3
• /
• pp.220-225
• /
• 2014

An efficient in vitro protocol has been established for somatic embryogenesis and plantlet conversion of Korean wild ginseng (Panax ginseng Meyer). Wild-type and mutant adventitious roots derived from the ginseng produced calluses on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 0.3 mg/L kinetin; 53.3% of the explants formed callus. Embryogenic callus proliferation and somatic embryo induction occurred on MS medium containing 0.5 mg/L 2,4-dichlorophenoxyacetic acid. The induced somatic embryos further developed to maturity on MS medium with 5 mg/L gibberellic acid, and 85% of them germinated. The germinated embryos were developed to shoots and elongated on MS medium with 5 mg/L gibberellic acid. The shoots developed into plants with well-developed taproots on one-third strength Schenk and Hildebrandt basal medium supplemented with 0.25 mg/L 1-naphthaleneacetic acid. When the plants were transferred to soil, about 30% of the regenerated plants developed into normal plants.

• Korean Journal of Plant Tissue Culture
• /
• v.27 no.4
• /
• pp.309-315
• /
• 2000

Panax ginseng is an important medicinal plant that has been used worldwide for geriatric, tonic, stomachic, and aphrodisiac treatments. Ginsenosides contained in the ginseng root are the main substances having active functions for human body. The price of ginseng is very expensive due to a complex process of cultivation, and the yield of ginseng is limited, which cannot meet the demand of the increasing market. Researchers have applied plant biotechnology to solve the problems but there are still things to be determined towards ginsenoside production by large-scale adventitious root culture. In this experiment, 5 to 20 liter bioreactors were employed to determine optimal conditions for adventitious root culture and ginsenoside production of Panax gineng. Callus was induced from the ginseng root on MS agar medium containing 1.0 mg. $L^{-1}$ 2,4-D and 0.1 mg. $L^{-1}$ kinetin. Then the callus was cultured on MS agar medium supplemented with 2.0 mg. $L^{-1}$ IBA, 0.1 mg. $L^{-1}$ kinetin, and 30 g. $L^{-1}$ to induce adventitious roots. The maximum root growth and ginsenoside production were obtained in 1/2 MS medium. 2.0 mg. $L^{-1}$ naphthalene acetic acid resulted in greater root growth than 2.0 mg $L^{-1}$ indole-3-butyric acid. Ginsenoside content increased with 2.0 mg. $L^{-1}$ benzyl adenin or kinetin. High concentrations of benzyl adenin (above 3.0 mg. $L^{-1}$ ) decreased the adventitious root growth and ginsenoside productivity. N $H_{4}$$^{+}$ inhibited the ginsenoside accumulation, while high concentrations of $K^{+}$, $Mg_{2}$$^{+}$, and $Ca_{2}$$^{+}$ increased it. N $H_{4}$$^{+}$ at 0.5 and 1.0 times of the normal amount in 3/4 SH medium resulted in the greatest biomass increase, but the highest ginsenoside productivity was obtained when N $O_{3}$$^{-}$ was used as the sole nitrogen source in the medium. Most microelements at high concentrations in the medium inhibited the root growth, but high concentrations of MnS $O_4$enhanced the root growth. Root dry weight increased with increasing sucrose concentrations up to 50 g. $L^{-1}$ , but decreased from 70 g $L^{-1}$ Ginsenoside productivity was maximized at the range of 20 to 30 g. $L^{-1}$ sucrose. In the experiment on bioreactor types, cone and balloon types were determined to be favorable for both adventitious root growth and ginsenoside production. Jasmonic acid was effective for increasing ginsenoside contents and Rb group ginsenosides mainly increased. These results could be employed in commercial scale bioreactor cultures of Panax ginseng.x ginseng.

• Journal of Plant Biotechnology
• /
• v.29 no.4
• /
• pp.247-252
• /
• 2002

The combination of radiation technique with an in vitro culture system was initiated to develop salt tolerant rice. We established an in vitro culture system to select tolerant lines against salt stress. NaCl tolerant cell lines were selected from the callus irradiated with gamma ray on N$_{6}$ medium with 1.5% NaCl and 2 mg/L 2,4-D. Regenerants (M$_1$) were obtained from the tolerant callus which was cultured for 30 days auxin-free medium. The M$_2$seeds were harvested from M$_1$plants on an individual plant basis. Thirty seedlings from each 450 M$_2$lines were transplanted in a field and total 5,000 M$_3$lines were harvested with an average 90 percent of fertile grain. M$_3$lines were utilized for selection of salt tolerance. Salinity-tolerant lines (225) were selected among 5,000 M$_3$lines. Of the 225 lines tested, the morphological traits of two lines (120-10 and -11) were far superior to control (Donagjinbyeo) in agromomic traits such as plant height, root length and no. of roots. Control and tolerant lines were analyzed by RAPD markers. Three polymorphic bands were presented in only tolerant lines, demonstrating a genetic difference between control and the tolerant lines. Such tolerant lines could be used as genetic resources to improve salt tolerance.e.

• Journal of Korean Society of Forest Science
• /
• v.75 no.1
• /
• pp.25-31
• /
• 1986

In order to contribute to the Korean tea-plant culture and tea industry by means of increasing the production of tea-plants, I have performed the tissue culture of the organs of the anther, leaf and stem. As for the culture-material, I have used the anther of tea (Thea sinensis) at the tetrad uninucleate microspore stage and used medium of modified Murashige and Skoog as the basal medium supplemented with the growth regulators of NAA and 2, 4-D, yeast, kinetin and others at various concentrations. As for the handling of material, I have followed the common methods of sterilization and microtoming and paraffine imbedding method and observed systematically periodic changes of the microspores in culture. I have divided the leaf, stem and root into segments and sterilized them and used the modified Murashige and Skoog as the basal medium and observed the differentiation of roots and callus and the results are as follows. 1. In case of anther, I have found 2n callus was found in 30 out of 100 segments in M2 medium. 2. The differentiation of roots appeared in 24.5% of total leaf segments cultured and in 50.5% of stem and in 43.9% of root. 3. When the differentiation of stem in different parts was observed, the most frequent differentiation was found in the second part of all the 4 parts. 4. The most frequent formation of callus was noticed from the anther-walls in case of anther culture and from the veins in case of leaf culture. It is concluded that the seedlings of tea-plant could be multiplied most by means of tissue culture of the second part of the tea-plant stem and reduction in the expenditures of tea-plant propagation was possible through tissue culture.

• Journal of Korean Society of Forest Science
• /
• v.78 no.4
• /
• pp.333-344
• /
• 1989

The objectives of this study were to investigate the growth of Gyrophora esculanta and to establish a method of tissue culture of the plant. The results obtained were as follows : 1. The Gyrophora esculanta was found growing mostly on the rock slopes of 722 m to 1915 min elevation on mountains in Korea. 2. Trees growing in the vicinity of the G. esculanta were mainly Quercus spp., Pinus thunbergii, Acer spp. and Lespedeza spp, Especially Quercus spp. was found growing in all of the study site. 3. The average Length of the rock slopes with G. esculanta growing on was 14 m and their aspects were mostly south. 4. The G. esculanta were found growing on rocks of Crystalline Schist, Quartz, Liparite, Granite, ete. Particularly they were mostly found on granites. The gradient of the rock slopes was in the range of 22-90 degrees. 5. The mean number of individuals of G. esculanta per one rock slope ranged from 14 at Mt. Bukhan to 70 at Mt. Jrri. Their mean diameter ranged from 1.8cm at Mt. Munsu to 4.6cm at Mt, Sokri. 6. The average percentage of G. esculanta with fruit body was 17.6%. The highest value was found at Mt. Cheonhwang (24.0%). 7. When the 100 segments of rhizoid of Gyrophora esculanta cultured in Detmer's medium supplemented with kinetine 5mg/l and 2, 4-D 3mg/l, n callus of microspore origins were induced from about 20% of the segments. As the induced n callus was transplanted on the six different types of rocks, it was observed that the juvenile G. esculanta grew best on granite and the development rate of G. esculanta on the granite was about 55%.

• Journal of Plant Biology
• /
• v.28 no.3
• /
• pp.233-241
• /
• 1985

The isolation and culture of protoplasts from hypocotyl-derived calluses of Glycine max (L.) Merr. cv. Jangyeop were obtained by digestion for 6 hrs in an enzyme solution containing 3.5% cellulase, 1.5% macerozyme, 10% sorbitol and 0.1% CaCl2.2H2O at pH 5.8. Newly formed cell wall of protoplasts cultured in MS agar medium containing 10 $\mu$M $\alpha$-naphthaleneacetic acid (NAA) and 32 $\mu$M N6-benzylaminopurine (BAP) could be observed after 24 hrs culture. The first cell division of the protoplasts was observed after 3 days of culture; cell clusters after 2 weeks of culture. When transferred to solid media, the protoplasts formed cell clusters gave rise to proliferating calluses.

• Journal of the Korean Society of Tobacco Science
• /
• v.11 no.2
• /
• pp.233-240
• /
• 1989

This experiment was carried out to select of heterokaryon based on the different buoyant densities of protoplasts. Protoplats were isolated from cultured cells (calli) of Nicotiana tobacum(cv. BY4) and from mesophyll cells of N. glauca. The two types of protoplats were fractionated by centrifugation in an iso-osmotic (770 mOs/kg. H2O) density gradients condition. Major difference in the buoyant density exists between two types of protoplasts isolated from different cells. The mesophyll protoplasts were fractionated in the higher gradient interphases than that of callus protoplasts. The two types of fractionated protoplasts were fused with 40% polyethylene glycol (PEG), and the protoplasts treated with PEG were separated by centrifugation in the same density gradients condition. The heterokaryons were fractionated in the intermediate density gradients.

• Journal of Marine Bioscience and Biotechnology
• /
• v.2 no.2
• /
• pp.77-80
• /
• 2007

Macroalgal tissues can be used as indicating materials for environmental assessment using several algal biotechnology techniques. As bioassay test organisms, macroalgal tissues are required as an axenic state for suitable biological indicators. Callus formation and blade regeneration under suitable culture conditions are also useful for the tests. Quantitative method using tetrazolium chloride or $alamarBlue^{TM}$ is devised on a rapid assessment of the seaweed viability. The use of RT-PCR especially differential display technique should provide the means for the detection and isolation of the responding genes induced by the environmental stress. Seaweed thriving in more environmental changes might contain more diverse biologically active substances.

• Journal of Plant Biotechnology
• /
• v.2 no.1
• /
• pp.35-41
• /
• 2000

Transgenic cabbage (Brassica oleracea ssp. capitata) plants resistant to the commercial herbicide Bast $a^{R}$ were obtained by Agrobacterium tumefaciens - mediated transformation. Hypocotyl segments of in vitro grown plants were infected with Agrobacterium tumefaciens LBA 4404 harboring plasmid pMOG6-Bar which contains hpt and bar genes. Explants were cultured on callus induction medium (MS basal medium + 1 mg/L NAA + 2 mg/L BA + 2 mg/L AgN $O_3$+ 100 mg/L carbenicillin + 250 mg/L cefotaxime) supplemented with 15 mg/L hygromycin. Hygromycin resistant calluses were transferred to shoot regeneration medium (MS basal medium + 0.1 mg/L NAA + 2 mg/L BA + 3% sucrose + 2 mg/L AgN $O_3$+ 15 mg/L hygromycin + 250 mg/L cefotaxime + 100 mg/L carbenicillin). In order to induce roots, elongated shoots were placed on the MS medium without plant growth regulators and hygromycin. Southern blot analysis of several putative transgenic plants indicated that one to five intact copies of Apt and bar genes were incorporated into the genome. Expression of bar gene was confirmed by Northern blot analysis and by herbicide resistant phenotype. Seed progeny from self-pollinated transformants expressed the herbicide resistance and showed Mendelian segregation of the introduced gene.e.

• Korean Journal of Environmental Agriculture
• /
• v.29 no.1
• /
• pp.7-11
• /
• 2010

This experiment was conducted to investigate the growth inhibition effects caused by continuous cropping soil in peony(Paeonia lactiflora Pallas). The effect of extracts from continuous cropping soil of peony was tested with bio-assay method using callus cells induced from peony filament tissues and seedlings derived from peony zygotic embryos. The cell viability and seedling growth were significantly inhibited by methanol extract in continuous cropping soil. Methanol extract from continuous cropping soil was successively fractionated with solvents such as n-hexane, ethyl acetate, n-butanol and water. The seedling growth was inhibited by ethyl acetate fraction obtained in methanol extract.