• Journal of Plant Biology
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• v.39 no.1
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• pp.71-77
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• 1996

Embryogenic callus was selected from callus induced from hypocotyl segment cultures of Daucus carota seedlings on MS basal medium supplemented with 1.0 mg/L 2,4-D. Cell clumps prepared from the embryogenic callus were transferred to MS medium without 2,4-D for somatic embryo development. Cotyledonary abnormalities were frequently observed on somatic embryos developed after the incubation of cell clumps in MS basal medium with 1.0 mg/L 2,4-D for one week and then subculture in the same medium but without 2.4-D for two weeks. The percentage of abnormalities was as follows: 5% one cotyledon, 21% three cotyledons, 6% four cotyledons, 5% five cotyledons, 0.2% six cotyledons and 1% trumpet-like cotyledons. On the other hand, the normal somatic embryo with two cotyledons appeared at 63%. The germination rate of somatic embryos was higher in two cotyledon somatic embryos than in multicotyledonary embryos. Trumpet-like somatic embryos did not germinate normally showing limited elongation and enlargement of roots and cotyledons without shoot development. From anatomical examination circular procambium in the root of somatic embryo began to branch around the middle regions of the hypocotyl which extended into the cotyledons through the cotyledonary nodes and the number of branched procambial strands in hypocotyl was equal to the number of cotyledons. Monocotyledonous somatic embryo always had larger cotyledon than that of somatic embryos with multicotyledons and had horseshoe-like cotyledons where the procambium was of the same structure.ucture.

• Korean Journal of Plant Tissue Culture
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• v.28 no.2
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• pp.103-108
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• 2001

The in vitro plantlets of cassava (Manihot esculenta Crantz cv. MColl 22) could be regenerated from nodal explant cultures in a liquid MS basal medium containing 0.01 mg/L zeatin for 2 weeks. The plantlets of 1.5∼2.5 cm in shoot length were transplanted to a glass bottle containing fine sand and acclimated under non-sterile conditions after excising their intact roots by: 1) prune leaving roots base of 1∼1.5 cm; 2) complete removal of roots; and 3) cutting off the rooting zone. The majority of in vitro-formed intact roots continued growth after transferred to soil, and all of the damaged roots stopped further growth. The plantlets with excised roots began to develop new roots within 7∼10 days after being transferred to a glass bottle, and a few of the pruned roots developed lateral roots from the remaining portion. Pruning and removal of in vitro roots resulted in a high survival rate (>87%), and did not significantly affect ex vitro root regeneration and acclimation, but the plantlets in which the rooting zone had been cut-off showed 73% survival rate. Pruning or removal of in vitro roots before transfer of plantlets is recommended for useful method of commercial micropropagation because of easier handling and high survival rate of plantlets.

• Korean Journal of Soil Science and Fertilizer
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• v.45 no.6
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• pp.1037-1042
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• 2012

An auxin-producing bacteria (2SJ8-02) was isolated from waste mushroom bed of Agaricus bisporus in Buyeo-Gun, Chungnam. The strain 2SJ8-02 was classified as a novel strain of Pantoea rwandensis based on a chemotaxanomic and phylogeneticanalyses. The isolate was confirmed to produce indole-3-acetic acid (IAA), one of auxin hormones, by TLC and HPLC analyses. The maximum concentration of IAA, $122mg\;L^{-1}$ was detected from the culture in R2A broth containing 0.1% tryptophan for 24 h at $35^{\circ}C$. The molecular weight of the main peak obtained by LC-mass analysis was 175 corresponding to that of IAA. To investigate the growth-promoting effects to the crops, the culture broth of Pantoea rwandensis 2SJ8-02 was infected to water cultures and seed pots of mung bean. In consequence, the adventitious root induction and root growth of mung bean were two times higher than those of the control.

• Journal of Plant Biotechnology
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• v.34 no.2
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• pp.139-144
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• 2007

Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.

• Journal of Korean Society of Forest Science
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• v.95 no.1
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• pp.32-37
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• 2006

The influence of different doses of jasmonic acid on both biomass of adventitious roots and accumulation of secondary metabolite in bioreactor cultures of Eleutherococcus koreanum was studied. The maximum growth (2.9 g Dry Weight/L) of adventitious roots was observed in the absence of jasmonic acid. The biomass of adventitious roots decreased with the increase in jasmonic acid dosage. High level of jasmonic acid efficiently stimulated the production of both eleutheroside B and E. The highest eleutheroside B ($49.0{\mu}g/g\;DW$) was observed at 0.4 mg/L jasmonic acid. Jasmonic acid at 0.2 mg/L led to the maximum accumulation of eleutheroside E ($64.7{\mu}g/g\;DW$). Whereas eleutheroside $E_1$ was maximally $19.0{\mu}g/g\;DW$ accumulated in the absence of jasmonic acid, the total eleutheroside ($259.9{\mu}g/L$) was the highest when 0.01 mg/L jasmonic acid was treated. In this study, the level of eleutheroside B decreased whereas the maximum levels of eleutheroside E and eleutheroside $E_1$ were observed at 10th and 4th day after jasmonic acid treatment.

• Journal of Plant Biotechnology
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• v.42 no.1
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• pp.34-42
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• 2015

This study was conducted to investigate the productivity of biomass and antioxidant compounds in Polygonum multiflorum by culturing explants in air-lift bioreactor containing Murashige and Skoog (MS) medium, by adding different concentrations of auxins [indole-3-butyric acid (IBA) and naphthalene acetic acid (NAA)], sucrose, methyl jasmonate (MeJA), and salicylic acid (SA). Results of this study revealed that the explants culturing on the medium supplemented with $9.84{\mu}M$ IBA and 50 g/L sucrose were observed to have higher productivity of biomass and bioactive compound than other treatments used. Thus, we expect that these results will be helpful for large-scale production of biomass and antioxidant compounds from Polygonum multiflorum.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.163-169
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• 2008

Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

• Proceedings of the Plant Resources Society of Korea Conference
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• 1999.10a
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• pp.46-57
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• 1999

Ginseng(Panax ginseng C.A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark(22$^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

• Journal of Mushroom
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• v.17 no.1
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• pp.12-18
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• 2019

An auxin-producing bacterium Yangsong-1 was isolated from a button mushroom bed in Chung cheongnam-do. The strain Yangsong-1 was classified as a novel strain of Arthrobacter enclensis based on a chemotaxonomic and phylogenetic analysis. The isolated A. enclensis Yangsong-1 was confirmed to produce indole-3-acetic acid (IAA), which is one of the auxin hormones. When the concentration of IAA was assessed by HPLC quantity analysis, the maximum concentration of IAA, $152.903mg\;L^{-1}$, was detected from the culture broth incubated in R2A medium containing 0.2% L-tryptophan for 48 h at $35^{\circ}C$. A negative relationship between IAA production and pH was estimated to show that the increase in IAA caused pH acidification of the culture. The effect of the supplement on L-tryptophan, a known precursor of IAA production, appeared to be at maximal production at 0.2% concentration and was rather reduced at concentration above 0.4%. To investigate the growth-promoting effects on the crops, the culture broth of A. enclensis Yangsong-1 was placed in water cultures and seed pots of mung beans and lettuce. In consequence, the adventitious root induction and root growth of mung beans and lettuce were 1.5 and 1.9 times higher, respectively, than those of the control.

• Journal of Plant Biotechnology
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• v.31 no.4
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• pp.295-300
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• 2004

This article was conducted to induce the transgenic hairy roots and determine the effect of culture conditions on optimum growth of hairy roots by Agrobacterium rhizogenes strain, 15834 in Lycium chinense Miller. Hairy roots of L. chinense Miller. were induced from leaf segments by co-cultivation with A. rhizogenes. When the hairy roots were cultured in various MS medium strength and sucrose concentrations, the highest growth of hairy roots was observed in half-strength MS media supplemented with 3% sucrose, respectively. In air lift bioreactor cultures, the liquid medium contained with 1/2 MS and 3% sucrose was also the best for optimum growth of hairy roots.