• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.41 no.4
• /
• pp.482-488
• /
• 1996

The experiment was carried out to the find variation of the sugar contents in the tip and basal part of the cotyledon and remaining portion of embryos in Phaseolus vulgahs seeds during germination with HPLC analysis method. Water content in cotyledon of kidney bean seed was about 6.4~6.5 of fresh weight and incresed to 45.8~71.2% during germination showing that tip part of cotyledon has more water content by 1.1~3.8% compared to the lower part of it. Higher water onten was observed in the rest parts of the seed except cotyledon such as plumule, radicle and hypocotyl showing that it increased to 72.2~93.3% depending on the different tissue organs. Main important sugars in kidney seeds during germination stages are; raffinose, sucrose, glucose and fructose, and the amount are differed with the kinds of embryo in kidney bean seed organs and stages of germination. Raffinose amount in kidney bean is increasing repeatly when seeds become wet but disappear it soon after seed have germinated especially in growing embryo parts. Raffinose in basal parts of cotyledons were still presented some an amount after germination. Sucrose is synthesized highly in plumule embryo at the beginning stage of germination but disappeared it from 5 days after seeding stages. Amount of sucrose in cotyledon of kidney seeds and seedlings increased continuously after germination. The amount of glucose and fructose in the cotyledons of kidney seeds during germination varied 5~10% or 5~15% but in the germinating and growing organs, plumule, they increased continuously after germination.

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

• Journal of Plant Biotechnology
• /
• v.40 no.4
• /
• pp.198-202
• /
• 2013

To produce transgenic cucumber expressing Nit gene coffering abiotic resistance, the cotyledonary-node explants of cucumber (cv. Eunsung) were inoculated with A. tumefaciens transformed with pPZP211 or pCAMBIA2300 carrying Nit gene, that has cis-acting element involved in resistance to various abiotic environmental stresses. After co-cultivation, the procedures of selection, shoot initiation, shoot elongation, and plant regeneration were followed by cotyledonary-node transformation method (CTM, Jang et al. 2011). The putative transgenic plants were selected when shoots were grown to a length greater than 3 cm from the cotyledonary-node explants on selection medium supplemented with 100 mg/L paromomycin as a selectable agent. The confirmation of transgenic cucumber was based on the genomic PCR, Southern blot analysis, RT-PCR, and Northern blot analysis. A 105 shoots (4.12%) selected from the selection mediums were obtained from 2,547 explants inoculated. Of them, putative transgenic plants were only confirmed with 45 plants (1.77%) by genomic PCR analysis. Transgenic plants showed that the Nit genes integrated into each genome of 39 plants (1.53%) by Southern blot analysis, and the expression of gene integrated into cucumber genome was only confirmed at 6 plants (0.24%) by RT-PCR and Northern blot analysis. These results lead us to speculate that the genes were successfully integrated and expressed in each genome of transgenic cucumber.

• Korean Journal of Medicinal Crop Science
• /
• v.14 no.4
• /
• pp.255-260
• /
• 2006

Culture conditions for high frequency plant regeneration via somatic embryogenesis from embryogenic cell suspension cultures of Hovenia dulcis are described. Germinated somatic embryos were selected for induction of secondary embryogenesis. Friable embryogenic cells were induced directly from somatic embryos when transfer to 1/3 MS solid or liquid medium lacking plant growth regulators. The temperature strongly effected on induction of secondary embryognesis than other conditions in culture. All somatic embryos produced friable embryogenic cell clumps within 10 days when germinated somatic embryos cultured in 1/3 MS medium at $30^{\circ}C$ in suspension culture. No somatic embryos formed from embryogenic cell suspension cultures at $18^{\circ}C$. Numerous somatic embryos were induced and subsequently developed uniformly into germination stage from suspended cell clumps after 4 weeks of culture on $18^{\circ}C$. Plantlets conversion were observed on $18^{\circ}C$ when germinated somatic embryos were transferred to 1/3 MS solid medium without plant growth regulators or supplemented with 0.1-0.5 mg/l benzyladenine.

• Journal of Plant Biotechnology
• /
• v.30 no.1
• /
• pp.47-52
• /
• 2003

Anthers from several lines of carrot (Daucus carota L.) were plated on the semi-solid B$_{5}$, basal medium supplemented with 2,4-D and NAA at two concentrations, 1.0 and 2.0 mg/L plus 0.2 mg/L BAP (benzylaminop-urine). Anthers of the most lines on the B$_{5}$ basal medium with 2,4-D showed higher percentages of callus formation than those with NAA. Particularly, in line 45477, highest percentages of callus formation (50%) were observed on B$_{5}$ medium with 1.0 mg/L 2,4-D plus 0.2 mg/L BAP. With 1.0 mg/L 2,4-D, two months was sufficient for initiation of callus development. Calli were regenerated into plantlets through embryogenesis onto regeneration medium without any growth regulators. When callus showing yellowish and soft structure was cultured, it yielded green plants at high regeneration rates, The response of anthers in callus induction and plant regeneration was different among lines investigated. Optimal callus induction and plant regeneration could be obtained through manipulating the concentration of growth regulators. Plantlets after transfer to perlite were grown successfully in greenhouse conditions. Anther culture of carrot will be used as a useful breeding tool in future.

• Journal of Plant Biotechnology
• /
• v.33 no.4
• /
• pp.271-276
• /
• 2006

Hypocotyl explants of Chinese cabbage (us. 'Jeong Sang' and 'Seoul') produced adventitious shoots on Murashige and Skoog (MS) basal medium supplemented with 4mg/L $AgNO_3$, 5 mg/L acetosyringone, 4 mg/L 6-benzyladenine and 3mg/L alpha-naphthaleneacetic acid (SI) after cocoultivation with strains of Agrobacterium tumefaciens (LBA4404) harboring the pCAMBIA1301 and the $_PPTN290$ containing hygromycin-resistance gene and paromomycin-resistance gene as a selectable marker genes, respectively. There was a significant difference in the frequency of transgenic plants depending on antibiotics and cultivars used. Paromomycin was better than hygromycin, and cultivar 'Jeong-sang' was higher than 'c.v. Seoul' in the frequency of transgenic plants. In particular, the highest frequency (0.70%) of transgenic plants was obtained from selection medium (SI) containing 100mg/L paromomycin in c.v., 'Jeong-sang' GUS positive response were obtained 9 plants and 3 plants from the cultivars, 'Jeong-sang' and 'Seoul', respectively. They were grown to maturity in a greenhouse and normally produced $T_1$ seeds. GUS histochemical assay for progeny $(T_1)$ revealed that the transgenes were expressed in the plant genome.

• Journal of Plant Biotechnology
• /
• v.42 no.1
• /
• pp.49-54
• /
• 2015

Based on the results in this study, here we propose a systematic micropropagation process for 'Gisela 5' that is one of the important dwarfing cherry rootstocks. When the apical tips detached from newly developed shoot in spring season were cultured on the half strength MS media with 0.5 mg/L IBA and 0.5 ~ 1.0 mg/L BA, the cultures scored the highest acquisition rate at 90% for normal shoot with vigorous growth and without hyperhydricity. As next step, the young shoots maintained in vitro well multiplied on the full strength MS medium supplemented with 0.5 mg/L IBA and 0.5 mg/L BA, in which multiplication rate was approximately nine-fold. Given the half strength MS medium containing 2.0 mg/L IBA, each transplanted shoot further developed robust roots. Finally, the plantlets were easily acclimatized in the compost consisted of vermiculite, perlite, and peatmoss in the proportion of 1:1:1. We expect that the results are useful for cherry cultivation and its rootstock production.

• Journal of Plant Biotechnology
• /
• v.38 no.1
• /
• pp.30-41
• /
• 2011

The influences of the agitation as well as the addition of medium during culture on the production of embryos were invested in isolated microspore culture of hot pepper (Capsicum annuum L.). When the culture medium was added during initial liquid culture step of liquid-double layer culture, the embryo yield and quality greatly increased. The most effective time point for medium addition was 5 days after the culture commenced. On the other hand, the effect of medium addition at later double layer culture step in liquid-double layer culture on the embryo production was less compared to that of medium addition during the initial liquid culture step. Agitating the culture for 1 week during later double layer culture step in liquid-double layer culture effectively increased the production of normal cotyledonary embryos. In the case of liquid culture, agitating the culture for 1 week from 7 days after the culture commenced was also effective for embryo development. However, when the total agitation time was longer (2 to 3 weeks) during liquid-double layer culture or liquid culture, the embryos developed abnormally in both cases. The normal cotyledonary embryos obtained in this study successfully developed to plants when transferred to regeneration media. These regenerated plants were either diploid or haploid, and there was a difference in the number of chloroplasts between guard cells of diploid and haploid. These results can be used as an important data for developing an efficient microspore culture system with high quality embryo production in hot pepper.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.31 no.1
• /
• pp.1-8
• /
• 2011

Molecular techniques such as genetic transformation are powerful tools that can be used for the genetic modification of warm-season grasses. The P. meyerianum with high regeneration ability was used for establishing an Agrobacterium-mediated transformation system. We investigated various factors affecting Agrobacterium infection by examining GUS gene expression of pCAMBIA1304 vector. Among various concentration of acetosyringone and betaine tested for inoculation and co-cultivation, 10 mg/L acetosyringone and 60 mg/L betaine resulted in the highest transformation frequency in terms of GUS expression. The calli of 4 species of Panicum spp. with excellent tissue culture response were inoculated with Agrobacterium under the optimal infection conditions. The high activity of GUS gene was observed in all species and hygromycin-resistant calli expressing GFP were obtained in P. meyerianum, P. longijubatum, P. stapfianum and guineagrass Noh-PL1. Co-cultivated calli were transferred onto the selection medium containing hygromycin, and the hygromycin resistant calli were selected after 3 months. Hygromycin-resistant plantlets were then successfully regenerated from the calli and grown in a greenhouse. We confirmed stable insertion of hpt gene among the hygromycin-resistant plantlets of P. meyerianum by PCR analysis.

• Journal of Plant Biotechnology
• /
• v.30 no.4
• /
• pp.381-385
• /
• 2003

An efficient method of plant regeneration from Acanthopanax chiisanensis somatic embryos was developed. Cotyledonary somatic embryos were obtained in liquid Murashige and Skoog (MS) medium from embryogenic cell suspension cultures. They were desiccated for 0 to 72 hr and then cultured on MS medium containing NAA, BA, GA$_3$, (0-0.5mg/L). The highest multiple shoots formation (100％) was obtained from 72 hr desiccated somatic embryos on ifs medium with 0.5mg/L NAA＋0.5mg/L BA or 0.5 mg/L NAA＋0.5mg/L BA＋0.5mg/L GA$_3$ after 6 weeks culture. Plant conversion from multiple shoots was not high. The highest plant conversion from multiple shoots was obtained on 1/3MS medium with 1.0mg/L GA$_3$. Converted plantlets were transferred to ex vitro condition and the highest survival rate (70％) of the plantlets was obtained on plastic pots containing vermiculite and sand. These results indicate that micropropagation procedure can be applied for an efficient mass propagation of Acanthopanax chiisanensis.