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Regulation of the Korean Radish Cationic Peroxidase Promoter by Phytohormones and Other Reagents

  • Lee, Dong-Ju;Kim, Sung-Soo;Kim, Soung-Soo
    • BMB Reports
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    • v.32 no.1
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    • pp.51-59
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    • 1999
  • The Korean radish cationic peroxidase (KRCP) promoter, comprising nucleotides -471 to +704 relative to the transcriptional initiation site, was fused to the GUS gene and transformed to tobacco BY-2 cells. We examined how auxin (2,4-dichlorophenoxyacetic acid, 2,4-D), cytokinin (6-benzylaminopurine, BAP), gibberellic acid ($GA_3$), abscisic acid (ABA), methyl jasmonate (MeJA), and phosphatidic acid (PA) affect the GUS expression in the presence or absence of 2,4-D in a modified LS medium. Exogenous 2,4-D or BAP greatly decreased the GUS expression regulated by the KRCP promoter in a modified LS medium containing 0.2 mg/l 2,4-D. $GA_3$ increased the GUS expression and ABA completely reduced the inductive effect of $GA_3$. The GUS expression was also increased dose-dependently by plant defense regulators, MeJA and PA. In contrast to the above results, auxin deprivation from the modified LS medium increased the GUS expression after treatment with exogenous 2,4-D whereas BAP still greatly decreased the GUS expression dose-dependently. $GA_3$ or MeJA slightly decreased the GUS expression. The data suggest that auxin deprivation changes the sensitivity of the suspension cells to exogenous chemicals and that the regulation of the KRCP promoter by 2,4-D, $GA_3$, and MeJA is dependent on auxin, whereas the regulation by BAP is not. This study will be valuable for understanding the function and expression mode of the Korean radish cationic peroxidase in Korean radish.

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Effects of Dessication, Sucrose and Salt Stress on the Regeneration of Portulaca oleracea Cultured Cells (건조, 염분 및 탕의 처리가 쇠비름(Portulaca oleracea L.) 배양세포의 재분화에 미치는 영향)

  • 권순태;오세명
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.2
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    • pp.117-121
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    • 1994
  • The optimal level of growth regulator for callus initiation stem explants was BAP 0.1mg/L combined with 2,4-D 1.0 mg/L in Murashige and Skoog (MS) medium supplemented with 30g/L sucrose and 10g/L agar, and that for cell growth was BAP 0.1+2,4-D 0.5 mg/L in MS liquid medium. The regeneration frequency of P. oleracea cells was significantly increased by subjecting the cells to dessication for 1and 2 h up to 83%, respectively, as compared with untreated control showing 61%. Cell viability and survival rate was inhibited by pretreatment of 0.6% NaCl for 2 days, while regeneration ability was not affected by the treatment. Pretreatment of 100g/L sucrose for 2 days markedly stimulated the regeneration of cells up to 81%. These results suggest that in addition to physiological changes, water stress resulted from dessication and high concentration of sucrose and NaCl is closely related to the regeneration of P. oleracea cultured cells.

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Effects of Mineral Media, Carbon Sources and Phytohormones on Micropropagation of Alnus hirsuta (물오리나무(Alnus hirsuta)의 기내증식에 미치는 기본배지, 탄소원 및 식물호르몬의 영향)

  • 김경희
    • Journal of Plant Biology
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    • v.35 no.2
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    • pp.135-142
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    • 1992
  • Shoot tip explants from germinated seeds of Alnus hirsuta were cultured on NT (Nagata and Takebe, 1971) mineral salts medium supplemented with 6% glucose, MS (Murashige and Skoog, 1962) vitamin mixture, polyvinylpyrrolidone (PVP) and $0-50\;\mu\textrm{M}$ 6-benzylaminopurine (BAP). Five $\mu\textrm{M}$ BAP was found to give the highest shoot multiplication rate. Accordingly about 200 shoots were obtained for further experiments by multiplying shoots on this medium for 4-5 months. Regardless of carbon sources, NT mineral medium produced 3-12 times of shoots than MS mineral medium did. On NT mineral medium, 3% sucrose, 3% glucose and 6% glucose yielded no significant differences. It was observed that media consisting of 1/4-1/2 strength NT mineral salts, 3% sucrose and $1-8\;\mu\textrm{M}$ IBA produced about 100% rooting rate. Almost 100% of the resulting plantlets survived after transfer to the soil by decreasing humidity stepwise.epwise.

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Hormonal Requirements Induced Different Regeneration Pathways in Alhagi graecorum

  • Hassanein, A.M.
    • Journal of Plant Biotechnology
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    • v.6 no.3
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    • pp.171-179
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    • 2004
  • Hormonal requirements inducing different regeneration pathways with particular emphasis on somatic embryo-genesis in Alhagi graecorum were studied. While combination of 0.5 $\mu{M}$ 2,4-dichlorophenoxyacetic acid (2,4-D), 2.5 $\mu{M}$ 6-benzylaminopurine (BAP) and 5 $\mu{M}$ 1-naphthaleneacetic acid (NAA) in MS medium induced callus formation and callus maintenance from internodal explants, each alone or in combination with other induced distinct regeneration pathway. Adventitious bud formation was induced on MS medium supplemented with 2.5 $\mu{M}$ BAP. It was improved when 2.5 $\mu{M}$ BAP was used in combination with 5 $\mu{M}$ NAA. MS medium containing 0.5 $\mu{M}$ 2,4-D or 5 $\mu{M}$ NAA induced the formation of abnormal direct somatic embryos. While increase of 2,4-D concentration (1.125-9) resulted in the formation of viable embryogenic mass, increase of NAA did not change its effect. NAA should be used in combination with 2,4-D even at low concentration (0.5 $\mu{M}$) to form embryogenic mass. In A. gaecorum, the role of 2,4-D as trigger of somatic embryogenesis and BAP as trigger of adventitious bud formation was deduced, but for maximum yield certain auxin-cytokinin ratio should be applied. Embryogenic masses characterized by high water content, low peroxidase activity, and low number of peroxidase and glutamate oxaloacetate transaminase bands in comparison with calli obtained under conditions stimulating adventitious bud formation. The resulted differential gene expression, which could be detected by native-PAGE patterns, could be used as marker for organogenic pathway in A. graecorum.

Plant Regeneration and Somatic Embryogenesis from Zygotic Embryo-derived Callus of Native Prunus yedoensis in Mt. Halla (한라산 자생 왕벚나무 접합자배 유래의 캘러스로부터 체세포배 형성과 식물체 재분화)

  • 고정군;박영철;양두영;김응식;오문유;고석찬
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.6
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    • pp.345-349
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    • 1997
  • Somatic embryos were induced through embryogenic callus derived from immature zygotic embryo culture of native Prunus yedoensis in Mt. Halla and regenerated into plantlets successfully. Embryogenic callus was induced most effectively on MS medium with 1.0 mg/L 2, 4-D and 0.1 mg/L BAP at an efficiency of approximately 60% using 45 day-old zygotic embryos after full blooming. Globular somatic embryos were induced from embryogenic callus on MS medium with 1.0 mg/L 2, 4-D and 0.1 mg/L BAP and these globular embryos developed to heart-shaped and cotyledonary embryos on hormone-free MS medium. Normal somatic embryos germinated 49% on 1/2 MS medium and the plants regenerated from the somatic embryos were morphologically normal.

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The Effects of Plant Growth Substances on the Callus Induction and Multiple Shoot Formation of Korean Lawngrass( Zoysia japonica Steud.) (식물생장조절제 처리가 들잔디의 Callus유기 및 Multiple Shoots형성에 미치는 영향)

  • 심재성;김동찬;서병기
    • Asian Journal of Turfgrass Science
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    • v.8 no.3
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    • pp.137-147
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    • 1994
  • We have established a high-frequency plant regeneration system via organogenesis from mature seed of Korean lawngrass(Zoysia japonica Steud.). The effects of 2,4-dichiorophenoxy acetic acid (2,4-D), 6-furfuryl amino purine (kinetin), $\alpha$-naphthaiene acetic acid (NAA), N6-benzyl amino pu-rine (BAP), and casein hydrolysate (CR) on cailus induction and multiple shoot formation on ex-posure to light were evaluated. Callus produced on the Murashige and Skoog (MS) medium containing 2,4-D and kinetin had high organogenesis potency. A single addition of 1.0 mg $L-^1$ 2,4-D significantly induced callus. Also, 1.0 mg L-$^1$ 2,4-D, with the addition of 0.1 mg $L-^1$ kinetin highly enlianced callus induction. The trend of cailus induction was also found on mediurn containing 0.1 mg $L-^1$ BAP with 1.0 mg $L-^1$2,4-D, and 1 g $L-^1$ CR with the addition of 1.0 mg $L-^1$ 2,4-D. However, NAA was no effective on callus formation. The growth of root was significantly high in the presence of 0.1 mg $L-^1$ kinetin compared to other concentrations. Over 2 mg $L-^1$ kinetin highly lengthened roots. Fresh weight of plantlet was highest on medium containing 0.1 mg $L-^1$ 2,4-D. Also, on medium containing 0.1 mg $L-^1$ BAP, fresh weight of piantlet was highly enhanced. BAP was significantly effective on multiple shoot formation, particularly when 2.0 mg $L-^1$ was added with 0.1 mg $L-^1$ 2,4-D. Callus induction and multiple shoot formation were achieved on MS basal medium containing 1.0 g $L-^1$ CH.

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Micropropagation of a Rare Species, Forsythia saxatilis N. through Tissue Culture (희귀(稀貴) 수종(樹種) 산개나리의 기내(器內) 번식(繁殖))

  • Moon, Heung Kyu;Suk, Gene Young;Kim, Sun Chang
    • Journal of Korean Society of Forest Science
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    • v.86 no.4
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    • pp.430-434
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    • 1997
  • Axillary bud explants from 3-year-old seedlings of Forsythia saxatilis N., rare and endangered species in Korea, were cultured on Murashige and Skoog's medium. The effect of various cytokinins(BAP, kinetin, and zeatin) at the different concentration(0.2, 0.5 and 1.0mg/L) was tested. Although an apparent shoot proliferation was not observed, zeatin showed slight promotional effect on normal shoot and leaf development. Both shoots and adventive roots could be induced simultaneously when the explants were cultured on the medium with kinetin, but adventive rooting was gradually reduced according as BAP and zeatin concentrations increased. Axillary shoot growth was promoted by the etiolation treatment. Shoot proliferation has been maintained more than three years with consecutive subculture. Rooted plantlets were successfully acclimatized in the artificial soil mixture and showed normal growth after transplantation into field.

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Effects of Growth Regulators on the Formation of Somatic Embryo and Adventitious Bud from the Cotyledon of Korean Ginseng (Panax ginseng C.A. Meyer) (고려인삼(Panax ginseng C.A. Meyer)의 자엽으로부터 체세포배 및 부정아의 발생에 미치는 식물호르몬의 영향)

  • Yang Deok-Chun;Yoon Eui-Soo;Choi Kwang-Tae
    • Journal of Ginseng Research
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    • v.23 no.4
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    • pp.199-204
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    • 1999
  • Cotyledon explants of Korean ginseng (Panax ginseng C.A. Meyer), a perennial medicinal plant, produced direct somatic embryos at a high frequency on MS medium without growth regulators. Cytokinin highly suppressed the somatic embryogenesis but stimulated direct fomlation of adventitious buds. BAP was more effective than kinetin for the formation of adventitious bud. IBA combination with cytokinin enhanced the frequency of adventitious bud formation. The highest frequency of adventitious bud formation were $40\%$ at 0.05 mg/l IBA and 5 mg/l BAP. Adventitious buds were mainly formed near the distal portion of cotyledon, while somatic embryos were only formed near the proximal portion of cotyledon. Adventitious buds were covered with sheath similar to axillary buds in the zygotic embryos, and then leaf-like epicotyls were developed.

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Plant Regeneration through Direct Somatic Embryogenesis from Immature Zygotic Embryo of Prunus yedoensis in Mt.Halla (한라산 자생 왕벚나무의 미성숙 접합자배로부터 직접 체세포배 발생을 통한 식물체 재생)

  • 고정군
    • Korean Journal of Plant Resources
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    • v.11 no.1
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    • pp.9-14
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    • 1998
  • Two types of somatic embryos were directly induced from the immature zygotic embryos of the wild prunus yedoensis in Mt. Halla after 16 weeks of culture on MS medium supplemented with 0.1mg/L $GA_3$ and 0.1mg/L BAPor 0.5mg/L $GA_3$ and 0.1mg/L BAP. One was normal single embryo with a single basal part. Normal somatic embryos germinated successfully on 1/2 MS medium. However, abnormal nulticotyledonary somatic embryos, formed shoots only on hormone free MS medium and about 80% of shoots rooted on MS medium with 0.5mg/L IBA. The mximum frequency (62.5%) of normal somatic embryos was directly obtained from the zygotic embryo 30 days after full blooming but it was decreased with further maturation.

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Effect of silver nitrate(AgNO3) and polyamines on shoot organogenesis and plant regeneration of Lycopersicon esculentum cultivar, Micro-Tom (질산은과 polyamines이 미니토마토, Micro-Tom 신초 기관발생과 식물체 재분화에 미치는 영향)

  • Kim, Yong-Ho;Park, Cheol-Ho;Park, Sang-Un
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.25-29
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    • 2002
  • The study was carried out to establish an efficient protocol for shoot organogenesis and plant regeneration from stem explant cultures of Lycopersicon esculentum cv. Micro-Tom. The regenerated shoots obtained from stem explant cultures on solid MS medium containing the different concentrations of BAP. The highest number of shoots (5.3) per explant and shoot growth (0.7 cm) was obtained on MS medium containing 4.0 mg/L BA. The additions of AgNO$_3$ and putrescine substantially improved the shoot regeneration frequency, at the optimal concentration of 7 mg/L and 50 mg/L respectively. The regenerated shoots (about 1 cm) were normal and could be easily rooted with 0.1 mg/L IBA treatment. The rooted plants were hardened and transferred to vermiculite with a 92% survival rate where they grew normally.