• Title/Summary/Keyword: In vitro roots

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High frequency somatic embryogenesis and plant regeneration of interspecific ginseng hybrid between Panax ginseng and Panax quinquefolius

  • Kim, Jong Youn;Adhikari, Prakash Babu;Ahn, Chang Ho;Kim, Dong Hwi;Kim, Young Chang;Han, Jung Yeon;Kondeti, Subramanyam;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.38-48
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    • 2019
  • Background: Interspecific ginseng hybrid, Panax ginseng ${\times}$ Panax quenquifolius (Pgq) has vigorous growth and produces larger roots than its parents. However, F1 progenies are complete male sterile. Plant tissue culture technology can circumvent the issue and propagate the hybrid. Methods: Murashige and Skoog (MS) medium with different concentrations (0, 2, 4, and 6 mg/L) of 2,4-dichlorophenoxyacetic acid (2,4-D) was used for callus induction and somatic embryogenesis (SE). The embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 Schenk and Hildebrandt (SH) medium. The developed taproots with dormant buds were treated with $GA_3$ to break the bud dormancy, and transferred to soil. Hybrid Pgq plants were verified by random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analyses and by LC-IT-TOF-MS. Results: We conducted a comparative study of somatic embryogenesis (SE) in Pgq and its parents, and attempted to establish the soil transfer of in vitro propagated Pgq tap roots. The Pgq explants showed higher rate of embryogenesis (~56% at 2 mg/L 2,4-D concentration) as well as higher number of embryos per explants (~7 at the same 2,4-D concentration) compared to its either parents. The germinated embryos, after culturing on $GA_3$ supplemented medium, were transferred to hormone free 1/2 SH medium to support the continued growth and kept until nutrient depletion induced senescence (NuDIS) of leaf defoliation occurred (4 months). By that time, thickened tap roots with well-developed lateral roots and dormant buds were obtained. All Pgq tap roots pretreated with 20 mg/L $GA_3$ for at least a week produced new shoots after soil transfer. We selected the discriminatory RAPD and ISSR markers to find the interspecific ginseng hybrid among its parents. The $F_1$ hybrid (Pgq) contained species specific 2 ginsenosides (ginsenoside Rf in P. ginseng and pseudoginsenosides $F_{11}$ in P. quinquefolius), and higher amount of other ginsenosides than its parents. Conclusion: Micropropagation of interspecific hybrid ginseng can give an opportunity for continuous production of plants.

Antifungal Activity of Root Colonizing Pseudomonas fluorescens MC07 is Responsible for Its Disease Suppression Ability (근권 정창 세균 Pseudomonas fluorescens MC07의 항진균 활성과 병 억제 능력)

  • 김진우;박병근;황인규;박창석
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.606-611
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    • 1998
  • An antagonistic bacterium, Pseudomonas flurorescens MC07 inhibited the mycelial growth of Rhizoctonia solani, Pythium ultimum, Fusarium oxysporum, and Phytophthora capsici in on potato dextrose agan (PDA) and other media. The strain MC07 conlonizes various plant roots and possesses antifungal activity. To determine the role of antifungal activity of the bacterium in disease suppression, a mutant Okm3-4 which lost its activity was isolated after screening 2,500 colonies generated by Omegon-Km insertions. The mutant Okm3-4 showed diminished growth inhibition of R. solani, P. ultimum, F. oxysporum, and Ph. capsici in vitro and had reduced suppressive effects on sesame damping.-off compared to the parental strain. In soils, accumulation of the pathogens by continuous cropping, 90% of sesame plants were killed by natural infection of damping-off whereas, only 29% of plants grown from seeds treated with MC07 were killed. On the other hand, 85% of plants died when sesame seeds were treated with the Okm3-4 cells. This indicated that antifungal activity of MC07 in vitro is directly responsible for the suppression of damping-off disease. Emergence rates of sesame seeds in pots containing diseased soil were 33%. However, MC07 treatments on seeds significantly improved emergence rates, which has similar effects of Benomyl treatment. The mutant Okm3-4 exhibited 53% of emergence rate. This indicated that antifungal activity of MC07 also affects the emergence rate of sesame seeds.

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In vitro Propagation using Shoot Tip Culture in Gold Tree[Dendropanax morbifera $L_{EV}]$. (황칠나무의 경정배양에 의한 기내번식)

  • 최성규;윤경원
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.6
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    • pp.464-467
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    • 2001
  • In order to establish a in vitro propagation system for gold tree[Dendropanax morbifera $L_{EV}$], the effects of auxins and cytokinins on shoot multiplication and rooting were investigated. Germination rate was the best in MS medium. The fresh weight and number of shoot were the best on the medium containing 0.1 or 1.0 mg/l BAP and 0.5 or 1.0 mg/l NAA. Shoots were successfully rooted in MS medium with 1.0 mg/l NAA. Roots were easily formed by the addition of auxins, especially 0.1 or 1.0 mg/l BAP.P.

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High Frequency Induction of Multiple Shoots from Nodal Explants of Vitex negundo L. Using Sodium Sulphate

  • Chandramu C.;Rao D. Manohar;Reddy V. Dashavantha
    • Journal of Plant Biotechnology
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    • v.5 no.2
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    • pp.107-113
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    • 2003
  • The effect of sodium sulphate on shoot induction and multiple shoot formation from nodal explants of Vitex negundo L. was tested on Murashige and Skoog's (MS) medium fortified with different auxins, cytokinins and sucrose. Highest percentage $(97.78\%)$ of explants for shoot induction and multiple shoot (20.68/explant) production were observed in the combination treatment of $N^6-Benzyl$ adenine (BA) $(17.80\;{\mu}M/L)$, ${\alpha}-Naphthalene$ acetic acid (NAA) $(2.15\;{\mu}M/L)$ and $5\%$ sucrose supplemented with 100 mg/L sodium sulphate. In vitro raised shoots were rooted on the half-strength MS medium fortified with different concentrations of NAA, Indole-3-acetic acid (IAA), and Indole-3-butyric acid (IBA) alone and in combinations. Among the treatments, $4.90\;{\mu}M/L$ of IBA was found most effective $(95.56\%)$ in inducing roots. The rooted plantlets were shifted to glasshouse for acclimatization and later transferred to the field with cent percent survival. Furthermore, in vitro flowering was observed in the shoots cultured on MS medium supplemented with BA $(8.90\;{mu}M/L)$ and NAA $(1.61\;{\mu}M/L)$.

Micropropagation of Aristolochia elegans(Mast.)

  • Lidia, Osuna T.;Alejandro, Mora I.;Elsa, Ventura Z.;Enrique, Jimenez F.;Crescencio, Bazaldua M.;Antonio, Jimenez A.
    • Journal of Crop Science and Biotechnology
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    • v.10 no.3
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    • pp.141-146
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    • 2007
  • The roots of Aristolochia elegans Mast.(Aristolochiaceae) are widely used in Mexican traditional medicine as a remedy for scorpion venom. Current experimental evidence supports its purported antidote properties. However, collection from the wilderness has lead to local extinction of natural populations. In order to contribute to species preservation, cultivation, and standardization of morphological and pharmacological properties, a micropropagation method was developed. This includes in-vitro germination of seeds to produce aseptic plantlets, induction of multiple budding, and acclimatization. The treatment with benzylamino purine(10 ${\mu}M$) induced the highest number of buds(3.1 on average) in both types of explants. On the other hand, indolebutyric acid(1.5 ${\mu}M$) caused the highest root index(11.8) per explant. One hundred percent of the micropropagated plantlets developed vigorously after the acclimatization process.

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Improved in vitro Regeneration of Potato (Solanum tuberosum cv. Superior) Transformed by Agrobacterium Expressing $\beta-Glucuronidase$

  • Park, Yoon-Kyung;Park, Gene-Sue;Yang, Young-Ki;Cheong, Hyeon-Sook
    • Journal of Plant Biology
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    • v.39 no.2
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    • pp.93-98
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    • 1996
  • In order to enhance the system of potato transformation and further regeneration, potato was transformed using the Agrobacterium tumefaciens harboring $\beta$-glucuronidase (GUS) gene. We found that a series fo modified medium ttained 100% shoot regeneration within 5 weeks after the preincubated explants on stage I medium were infected with Agrobacterium. Callus appeared at the cut edges of stem segments on stage II medium, mainly at the basal parts. Some explants started to form shoots after two to three weeks on stage III medium containing kanamycin (50 mg/L). When transferred to MS medium containing 200 mg/L kanamycin, 81% of the transformed shoots formed roots at the cut edge of the plantlets. In contrast, untrasformed shoots never rooted and became yellowish after few weeks under the same conditions. Southern and northern analysis indicated in vitro shoot regeneration on the callus derived from the potato explants, which were incubated with Agrobacteria. The regeneration cycle was shortened after the transformatin and finally the transformation efficiency was highly enhanced.

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Effect of Acidic Polysaccharide Components of Korean Ginseng on Lipolytic Action of Toxohormone-L and on Activity of Angiotensin Converting Enzyme (고려인삼중 다당체 성분이 암독소 호르몬-L의 지방분해 작용과 안지오텐신 변환효소의 활성에 미치는 영향)

  • Lee, Sung-Dong;Hwang, Woo-Ik;Okuda, Hiromichi
    • Journal of Ginseng Research
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    • v.20 no.3
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    • pp.248-255
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    • 1996
  • This study was devised to observe in vitro, the inhibitory effects of acidic polysaccharide fractions from Korean red ginseng (KRG) and white ginseng (KWG) on the lipolytic action of loxohormone-L and on angiotensin converting enzyme (ACE, peptidyldipeptidase hydrolase, EC 3.4.15.1) . The crude acidic polysaccharides (CAP) extracted from main and lateral roots of KRG and KWG were separately purified through several procedures. The total inhibitory activities on the lipolytic action of toxohormone-L of CAP from main roots of KRG and KWG was higher than those of CAP from lateral roots of KRG and KWG, respectively, and that of CAP from main root of KRG was 3.1 times higher than that of CAP from main root of KWG. The specific activity of CAP from main root of KRG was measured as 5.40 units/mg, when one unit was defined as the amount giving 50% inhibition on toxohormone-L induced lipolysls. A subfraction named PG4 3 obtained by replanted chromatography on DEAE-TOYOPEARL 650M gave the specific activity of 24.4 units/mg. On the other hand, it was found that the total inhibitory activity on ACE of CAP from lateral root of KRG was the highest among the 4 kinds of CAP, but the specific activity of CAP from lateral root of KWG was the highest.

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In Vitro Anticomplementary Activity of Hederagenin Saponins Isolated from Roots of Dipsacus asper

  • Oh, Sei-Ryang;Jung, Keun-Young;Son, Kun-Ho;Park, Si-Hyung;Lee, Im-Seon;Ahn, Yung-Seop;Lee, Hyeong-Kyu
    • Archives of Pharmacal Research
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    • v.22 no.3
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    • pp.317-319
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    • 1999
  • Anticomplementary activity of hederagenin and related saponins isolated from Dipsacus asper was investigated in vitro. HN saponin F (3) was most potent with $IC_{50}$ value of$ 3.7{\times}10^{-5} M$ followed by 3-O-${\beta}-D-glucopyranosyl-(1{\rightarrow} 3)-{\alpha}-L-rhamnopyranosyl-(1{\rightarrow}2)-{\beta}-L-arabinopyranosyl$ hederagenin $28-O-{\beta}-D-glucopyranosyl-(1{\rightarrow}6)-beta$-D-glucopyrano side (8), $3-O-{\beta}-L-arabinopyranosyl$ hederagenin $28-O-{\beta}-D-glucopyranosyl-(1{\rightarrow}6)-{\beta}-D-glucopyranoside$ (5), dipsacus saponin A (4), and hederagenin (1) on the classical pathway (CP) of complement system, while the saponins 3-5 did not show the inhibition of hemolysis and rather increase the hemolysis on the alternative pathway (AP). However, all of C-3 monodesmosides [prosapogenin CP (2), dipsacus saponin B (6), and dipsacus saponin C (7)] evoked hemolysis directly on the erythrocytes.

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Genotoxicity and Interference with Cell Cycle Activities by an Ethanolic Extract from Thai Plumbago indica Roots in Human Lymphocytes in vitro

  • Thitiorul, Sumon;Ratanavalachai, Treetip;Tanuchit, Sermkiat;Itharat, Arunporn;Sakpakdeejaroen, Intouch
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2487-2490
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    • 2013
  • In Thai traditional medicine, Plumbago indica or Jetamul-Pleung-Dang in Thai is known to have health benefit especially for anti-inflammatory, antibacterial, and antitumor activities. However, the mechanisms of its action are still uncertain. One of which might be genotoxic effects. In the present study, we investigated the genotoxicity of an ethanolic extract of Plumbago indica root (EEPIR) by sister chromatid exchange (SCE) assay in human lymphocytes. Results have shown that all treatments with EEPIR ($12.5-100{\mu}g/ml$) could induce cell cycle delay as shown by significant increase in the number of metaphase cells in the first cell cycle but neither in the second nor the third cell cycle. Only at concentrations of 25, 50, and $100{\mu}g/ml$ were SCE levels significantly increased above that of the control (p<0.05). EEPIR at a concentration of $500{\mu}g/ml$ induced cell death as few mitotic cells were shown. Accordingly, EEPIR ($25-100{\mu}g/ml$) is genotoxic in human lymphocytes and cytotoxic at concentrations of ${\geq}500{\mu}g/ml$ in vitro. Therefore, these activities of the EEPIR could serve its potential therapeutic effects, especially as an anticancer agent. Further study of EEPIR in vivo is now needed to support this in vitro evidence.

In Vitro and In Vivo Inhibitory Effects of Gaseous Chlorine Dioxide Against Diaporthe batatas Isolated from Stored Sweetpotato

  • Lee, Ye Ji;Jeong, Jin-Ju;Jin, Hyunjung;Kim, Wook;Yu, Gyeong-Dan;Kim, Ki Deok
    • The Plant Pathology Journal
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    • v.35 no.1
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    • pp.77-83
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    • 2019
  • Chlorine dioxide ($ClO_2$) can be used as an alternative disinfectant for controlling fungal contamination during postharvest storage. In this study, we tested the in vitro and in vivo inhibitory effects of gaseous $ClO_2$ against Diaporthe batatas SP-d1, the causal agent of sweetpotato dry rot. In in vitro tests, spore suspensions of SP-d1 spread on acidified potato dextrose agar were treated with various $ClO_2$ concentrations (1-20 ppm) for 0-60 min. Fungal growth was significantly inhibited at 1 ppm of $ClO_2$ treatment for 30 min, and completely inhibited at 20 ppm. In in vivo tests, spore suspensions were drop-inoculated onto sweetpotato slices, followed by $ClO_2$ treatment with different concentrations and durations. Lesion diameters were not significantly different between the tested $ClO_2$ concentrations; however, lesion diameters significantly decreased upon increasing the exposure time. Similarly, fungal populations decreased at the tested $ClO_2$ concentrations over time. However, the sliced tissue itself hardened after 60-min $ClO_2$ treatments, especially at 20 ppm of $ClO_2$. When sweetpotato roots were dip-inoculated in spore suspensions for 10 min prior to treatment with 20 and 40 ppm of $ClO_2$ for 0-60 min, fungal populations decreased with increasing $ClO_2$ concentrations. Taken together, these results showed that gaseous $ClO_2$ could significantly inhibit D. batatas growth and dry rot development in sweetpotato. Overall, gaseous $ClO_2$ could be used to control this fungal disease during the postharvest storage of sweetpotato.