• Title/Summary/Keyword: Indole-3-acetic acid (IAA)

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Metabolic Engineering of Indole Glucosinolates in Chinese Cabbage Plants by Expression of Arabidopsis CYP79B2, CYP79B3, and CYP83B1

  • Zang, Yun-Xiang;Lim, Myung-Ho;Park, Beom-Seok;Hong, Seung-Beom;Kim, Doo Hwan
    • Molecules and Cells
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    • v.25 no.2
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    • pp.231-241
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    • 2008
  • Indole glucosinolates (IG) play important roles in plant defense, plant-insect interactions, and stress responses in plants. In an attempt to metabolically engineer the IG pathway flux in Chinese cabbage, three important Arabidopsis cDNAs, CYP79B2, CYP79B3, and CYP83B1, were introduced into Chinese cabbage by Agrobacterium-mediated transformation. Overexpression of CYP79B3 or CYP83B1 did not affect IG accumulation levels, and overexpression of CYP79B2 or CYP79B3 prevented the transformed callus from being regenerated, displaying the phenotype of indole-3-acetic acid (IAA) overproduction. However, when CYP83B1 was overexpressed together with CYP79B2 and/or CYP79B3, the transformed calli were regenerated into whole plants that accumulated higher levels of glucobrassicin, 4-hydroxy glucobrassicin, and 4-methoxy glucobrassicin than wild-type controls. This result suggests that the flux in Chinese cabbage is predominantly channeled into IAA biosynthesis so that coordinate expression of the two consecutive enzymes is needed to divert the flux into IG biosynthesis. With regard to IG accumulation, overexpression of all three cDNAs was no better than overexpression of the two cDNAs. The content of neoglucobrassicin remained unchanged in all transgenic plants. Although glucobrassicin was most directly affected by overexpression of the transgenes, elevated levels of the parent IG, glucobrassicin, were not always accompanied by increases in 4-hydroxy and 4-methoxy glucobrassicin. However, one transgenic line producing about 8-fold increased glucobrassicin also accumulated at least 2.5 fold more 4-hydroxy and 4-methoxy glucobrassicin. This implies that a large glucobrassicin pool exceeding some threshold level drives the flux into the side chain modification pathway. Aliphatic glucosinolate content was not affected in any of the transgenic plants.

Auxin and Cytokinin Affect Biomass and Bioactive Compound Production from Adventitious Roots of Eleutherococcus koreanum (섬오갈피 부정근 배양 시 오옥신과 사이토키닌이 생장과 생리활성물질 생산에 미치는 영향)

  • Lee, Eun-Jung;Kim, Myong-Ki;Paek, Kee-Yoeup
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.678-684
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    • 2010
  • In an attempt to improve biomass and bioactive compound production, we cultured adventitious roots of $Eleutherococcus$ $koreanum$ in 250 mL Erlenmeyer flasks using Murashige and Skoog (MS) medium with different concentrations of auxins (IBA, NAA, IAA) and cytokinins (BA, kinetin, TDZ). Root biomass (fresh and dry weight) was enhanced at $5mg{\cdot}L^{-1}$ indole-3-butyric acid (IBA) after 5 weeks of culture. The content of total phenolics and flavonoids was also increased with $5mg{\cdot}L^{-1}$ IBA compared to ${\alpha}$-naphtalene acetic acid (NAA) or indole-3-acetic acid (IAA) treatments. The combination of $5mg{\cdot}L^{-1}$ IBA with $0.1mg{\cdot}L^{-1}$ thidiazuron (TDZ; N-phenyl-N'-1,2,3,-thidiazol-5-ylurea) enhanced root fresh and dry weight (1.4- and 1.6-fold, respectively) as well as the content of total phenolics and flavonoids compared to the relative control (without cytokinin). On the contrary, $N_6$-benzyladenine (BA) and 6-furfurylaminopurine (kinetin) did not significantly affect root biomass and bioactive compound production in adventitious roots of $E.$ $koreanum$. These results suggested that $5mg{\cdot}L^{-1}$ IBA combination with $0.1mg{\cdot}L^{-1}$ TDZ supplementation was most suitable for both biomass and bioactive compound production from adventitious roots of $E.$ $koreanum$.

In vitro propagation of Bambusa nutans Wall. ex Munro through axillary shoot proliferation

  • Negi, Divya;Saxena, Sanjay
    • Plant Biotechnology Reports
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    • v.5 no.1
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    • pp.35-43
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    • 2011
  • This communication describes for the first time an efficient and reproducible protocol for large-scale multiplication of Bambusa nutans. Nodal segments collected from field-grown clumps and cultured on Murashige and Skoog (MS) medium supplemented with $4.4{\mu}M$ benzylaminopurine (BA) and $2.32{\mu}M$ kinetin (Kin) gelled with 0.2% gelrite yielded 80% aseptic cultures with 100% bud-break. The in vitro-formed shoots obtained after bud-break were successfully multiplied in MS liquid medium supplemented with $13.2{\mu}M$ BA, $2.32{\mu}M$ Kin, and $0.98{\mu}M$ indole-3-butyric acid (IBA). Sub-culturing of shoots every 3 weeks on fresh multiplication medium yielded a consistent proliferation rate of 3.5-fold. Shoot clusters containing three to five shoots were successfully rooted with 100% success on half-strength MS liquid medium supplemented with $9.8{\mu}M$ IBA, $2.85{\mu}M$ indole-3-acetic acid (IAA), $2.68{\mu}M$ naphthaleneacetic acid (NAA), and 3% sucrose. Plantlets grown in vitro were acclimatized and subsequently transferred to the field. Inter-simple sequence repeat analysis has confirmed the genetic uniformity of the tissue-cultured plants up to 27 passages.

Change of Sprouting-related Enzymes Activities and Food Quality Characteristics of Sweetpotato Root (Ipomea batatas Lam.) by Electron Beam Irradiation (전자빔 조사에 의한 고구마의 발아관련 효소의 활성과 식품특성 변화)

  • Lim, Sung Jin;Song, Mi Seon;Lee, Gyeong Ae;Cho, Jae-Young
    • Journal of Applied Biological Chemistry
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    • v.55 no.4
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    • pp.267-272
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    • 2012
  • We investigated that electron beam irradiation is the effective method to control the sprouting of sweetpotato roots without changing of food quality characteristics. In 12 and $25^{\circ}C$ storage after electron beam irradiation, all control samples were sprouted from 6 and 4 weeks after storage, respectively. The sprouting rate of control increased with time and the rate reached to 11.2-12.4 and 70.5-74.2% at 8 weeks after 12 and $25^{\circ}C$ storage. Also, the sprouting of middle and below positioning sweetpotato roots at 12 and $25^{\circ}C$ storage after irradiation reached to 8.6-11.3 and 42.7-48.7% after a storage period of 8 weeks, respectively. However, the sprouting of all sweetpotato roots stored at $4^{\circ}C$ and upper (0-7 cm) positioning samples of box stored at 12 and $25^{\circ}C$ with electron beam was completely inhibited due to increase peroxidase and indole acetic acid (IAA) oxidase activity. Also, all samples with electron beam such as hardness, pH, sugar content, weight loss, and vitamin C and dacarotene content did not differ from that of the control. Therefore, if electron beam will be irradiated to sweetpotato roots above 0.1 kGy before packing, it will effectively inhibit their sprouting stored at $25^{\circ}C$ without the change of food quality characteristics.

Effect of Ascorbic Acid on the Gravitropic Response of Primary Roots in Maize (옥수수 일차뿌리에서 Ascorbic acid가 굴중성 반응에 미치는 효과)

  • Kim, Chung Su;Mulkey, Timothy J.;Kim, Soon Young
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1364-1370
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    • 2014
  • Ascorbic acid (AA) is a multifunctional metabolite in plants that is essential for plant development and growth. We examined the effect of AA, an antioxidant, on the gravitropic response of primary roots in maize. The application of $10^{-3}$ M AA to the elongation zone did not affect the gravitropic response and slightly inhibited the root growth. However, treatment with both $10^{-5}$ M and $10^{-3}$ M AA at the root tip increased the gravitropic response and inhibited root growth. Differences in indole-3- acetic acid (IAA) activity between the upper and lower hemispheres of the root resulted in differential elongation along the horizontal root. Roots are extremely sensitive to IAA, and increasing the amount of IAA in the lower hemisphere of the root inhibited elongation. Therefore, we examined the effect of IAA in the presence of AA. The inhibitory effect of AA on the gravitropic response was greater in combination with IAA. To understand the role of AA in the regulation of root growth and the gravitropic response, we measured ethylene production in the presence of AA in the primary roots of maize. AA stimulated ethylene production via the activation of the 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase gene, which regulates the conversion of ACC to ethylene. These results suggest that AA alters the gravitropic response of maize roots through modification of the action of ethylene.

Involvement of Pyridoxine/Pyridoxamine 5′- Phosphate Oxidase (PDX3) in Ethylene-Induced Auxin Biosynthesis in the Arabidopsis Root

  • Kim, Gyuree;Jang, Sejeong;Yoon, Eun Kyung;Lee, Shin Ae;Dhar, Souvik;Kim, Jinkwon;Lee, Myeong Min;Lim, Jun
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1033-1044
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    • 2018
  • As sessile organisms, plants have evolved to adjust their growth and development to environmental changes. It has been well documented that the crosstalk between different plant hormones plays important roles in the coordination of growth and development of the plant. Here, we describe a novel recessive mutant, mildly insensitive to ethylene (mine), which displayed insensitivity to the ethylene precursor, ACC (1-aminocyclopropane-1-carboxylic acid), in the root under the dark-grown conditions. By contrast, mine roots exhibited a normal growth response to exogenous IAA (indole-3-acetic acid). Thus, it appears that the growth responses of mine to ACC and IAA resemble those of weak ethylene insensitive (wei) mutants. To understand the molecular events underlying the crosstalk between ethylene and auxin in the root, we identified the MINE locus and found that the MINE gene encodes the pyridoxine 5′-phosphate (PNP)/pyridoxamine 5′-phosphate (PMP) oxidase, PDX3. Our results revealed that MINE/PDX3 likely plays a role in the conversion of the auxin precursor tryptophan to indole-3-pyruvic acid in the auxin biosynthesis pathway, in which TAA1 (TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1) and its related genes (TRYPTOPHAN AMINOTRANSFERASE RELATED 1 and 2; TAR1 and TAR2) are involved. Considering that TAA1 and TARs belong to a subgroup of PLP (pyridoxal-5′-phosphate)-dependent enzymes, we propose that PLP produced by MINE/PDX3 acts as a cofactor in TAA1/TAR-dependent auxin biosynthesis induced by ethylene, which in turn influences the crosstalk between ethylene and auxin in the Arabidopsis root.

Proliferation, Accumulation of Polyphenols, and Antioxidant Activities of Callus from the 'Anji Baicha' Cultivar of Tea [Camellia sinensis (L.) O. Ktze.]

  • Liu, Mingfei;Wang, Junli;Tian, Birui;Huang, Jingjing;Zhang, Rongrong;Lin, Yuxing;Xiao, Zefeng
    • Horticultural Science & Technology
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    • v.35 no.2
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    • pp.252-264
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    • 2017
  • Tea is one of the most consumed beverages worldwide and the relatively high levels polyphenols is benefit for health. In this study, we developed an efficient system for proliferation of callus from 'Anji Baicha', a cultivar of tea (Camellia sinensis). Callus tissue was initially induced by culturing leaf explants on medium containing different plant growth regulators. For callus induction, thidiazuron (TDZ) was more effective than 2,4-dichlorophenoxyacetic acid (2,4-D), ${\alpha}-naphthalene$ acetic acid (NAA), and $N^6-benzyladenine$ (BA). The frequency of callus induction from leaf explants reached 90.21% on $1.0mg{\cdot}L^{-1}$ TDZ and the developed callus was reddish and friable. We also tested the effect of different concentrations of NAA, 2,4-D, indole 3-acetic acid (IAA), BA, and TDZ, alone and in combinations, on callus proliferation. Medium supplemented with TDZ in combination with IAA was suitable for callus proliferation and accumulation of tea polyphenols. The growth index value and tea polyphenol content of callus cultured on MS medium containing $0.5mg{\cdot}L^{-1}$ TDZ and $1.0mg{\cdot}L^{-1}$ IAA was maximally 1,351% and 23.24%, respectively, and the relative abundance of epicatechin was as high as 17.449%. We also measured the antioxidant activity of all samples and the callus with the highest tea polyphenol content also exhibited high potential radical scavenging activity.

Control of In Vitro Flowering of Ginseng(Panax ginseng C. A. Meyer) by Growth Regulators (생장조절제에 의한 인삼(Panax ginseng C. A. Meyer)의 기내 화아형성 조절)

  • 이행순
    • Journal of Plant Biology
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    • v.32 no.4
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    • pp.255-264
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    • 1989
  • Ginseng zygotic embryos, seedlings, and exised cotyledonary nodes were cultured on Murashinge and Skoog's(MS) medium, supplemented with 6-benzyladenine(BA) and gibberellic acid(GA3) to induce flower buds. As the concenteration of nitrogen compounds in MS medium was reduced to half of its strength, the flowering frequency of zygotic embryos increased up to 90%. The optimum concentration of sucrose in the medium for flowering of seedlings was 30-60 g/1. In all cases flower buds were formed on elongated axillary branches from the cotyledonary nodes, while the apices remained vegetative. When zygotic embryos and excised cotyledonary nodes were cultured on the medium, supplemented with all possible combinations of BA, GA3, and abscisic acid(ABA) of 5 $\mu$M indole-3-acetic acid(IAA) in the above combinations did not affect flowering. These results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respectively, in the induction of flowering of ginseng.

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Establishment of Cell Suspension Cultures and Plant Regeneration in White Dandelion (Taraxacum coreanum NAKAI.)

  • Sun, Yan-Lin;Kim, Jae-Hak;Hong, Soon-Kwan
    • Korean Journal of Plant Resources
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    • v.24 no.3
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    • pp.280-285
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    • 2011
  • In this study, we established a novel somatic embryogenesis and plant regeneration system through cell suspension culture of white dandelion (Taraxacum coreanum NAKAI.). Embryogenic calli could be initiated from leaf and root explants of sterile seedlings on solid Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) after 3-week cultures. To proliferate embryogenic calli rapidly, cell suspension culture was performed with transferred to liquid MS medium with various combinations of plant growth regulators (PGRs) including 2,4-D, ${\alpha}$-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA), $N^6$-benzylamino purine (BAP), thidiazuron (TDZ), and kinetin. During suspension cultures, embryogenic calli not only greatly proliferated, but shoot organogenesis also simultaneously occurred from the surface of somatic embryos. Among them, TDZ at lower concentration, 0.1 mg/L produced the highest efficiency of somatic embryo formation and shoot organogenesis. Rooting of embryogenic calli with adventitious shoots was done on solid MS medium containing 0.1 mg/L NAA and 0.3% activated carbon. Nearly 80% of embryogenic calli with shoot organogenesis could be rooted normal. Well-rooted plantlets were transferred into pots under a greenhouse condition, and plants derived from this system appeared phenotypically normal.

Studies on Multiplication of Cornus of officinalis by in vitro Culture I. Callus Induction, Shoot Propagation and Root Differentiation through Bud Culture (산수유(山茱萸)(Cornus officinalisis)의 기내증식(器內增殖)에 관한 연구(硏究) I. 액아배양(腋芽培養)에 의한 Callus 유기(誘起), Shoot 증식(增殖) 및 뿌리 분화(分化))

  • Park, Chung-Heon;Seong, Nak-Sul;Lee, Seung-Tack;Youn, Kyu-Bok;Son, Su-Gyu
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.1
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    • pp.63-69
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    • 1993
  • Present experiment were attempted to examine in vitro multiplication throughbud culture of Cornus officinalis. Bud derived shoot formation was established successfully on Murashige and Skoog's medium supplemented with $0.5mg\;/\;{\ell}$ BAP(N-benzyl amino purine). The shoot proliferation increased on the Driver Kuniyuki Walnut medium containing $0.5mg\;/\;{\ell}$ NAA(Napthalene acetic acid) and $0.5mg\;/\;{\ell}$ BAP. Addition of 2,4-D(2,4-Dichlorophenoxy acetic acid) to the media produced excessive callus inducton. IAA(Indole-3-acetic acid) and IBA (Indole-3-bu-tyric acid) enhanced multple shooting, and NAA showed callus induction and multiple shooting. Shoot growth was enhanced supplemented with 3% sucrose, $2g\;/\;{\ell}$ activated charcoal, and 1 / 4MS in organic salts. However, root formation of proliferated shoots was low about 5%

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