• Title/Summary/Keyword: IAA biosynthesis

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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.

Analysis and Culture Conditions for Biosynthesis of Polyacetylene from Callus of Ginseng Superior lines (인삼 육성계통 캘러스부터 Polyacetylene의 분석 및 생합성에 미치는 배양조건)

  • 양덕춘;송남현;양계진;배창휴
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.123-128
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    • 2001
  • In order to develop the biotechnological methods for the mass production of anticancer compounds from tissue culture of Panax ginseng C.A. Mayer, these studies were carried out for the selection of ginseng cell lines containing higher concentration of polyacetylene compounds and optimal condition for their biosynthesis. Panaxynol, one of ginseng polyacetylene, was not detected in any callus induced from ginseng superior cell lines cultured on MS medium supplemented with $\beta$-chlorophenoxy acetic acid (CPA). Panaxydol, another one of polyacetylene and anticancer compounds, were detected in calli of 5 cell lines by thin layer chromatogram and gas chromatogram. Among the 18 ginseng superior lines, the cell line 30201 has higher content of panaxydol. Especially, panaxydol was not detected in the callus induced from cell line 10301 which cultured on the medium containing CPA only, however, it was detected on the same callus cultured on mixed medium containing CAP 2 mg/L and BA 0.05 mg/L. SH medium was better than MS medium for ginseng callus growth and biosynthesis of polyacetylene, and also found that it was not effected by NAA and sucrose concentration in the culture medium.

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Effects of Plant Growth Regulators on Physiology of Germinating Punax ginseng Seed (식물 생장조절제가 인삼 (Punax ginseng) 종자의 발아생리에 미치는 영향)

  • Eung-Ryong Son;Won-Mok Park;C. Pertzsch
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.24 no.1
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    • pp.99-106
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    • 1979
  • The undehiscent ginseng seed did not germinate, even if the seeds were treated with GA_3, kinetin or IAA. Only GA_3 stimulated germination of dehiscent ginseng seed. The physiological roles of gibberellic acid on stimulation of the seed germination were enhancing production of soluble carbohydrate and sucrose. Then gibberellic acid stimulated biosynthesis of insoluble cellural materials and amino acids from sugars and incorporation of amino acids into protein. The fruit coat of ginseng seed did not impede water imbibition, but did function as water absorbor and reservoir.

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Kinetic Analyses of Spermine Effects on Petiole Elongation in Ranunculus sceleratus

  • Chang, Soo-Chul
    • Journal of Plant Biology
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    • v.37 no.4
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    • pp.397-402
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    • 1994
  • Possible roles of polyamines in the inhibition of cell elongation in Ranunculus petioles were investigated. Exogenously apoplied polyamines greatly inhibited the auxin-induced petiole growth, while treatment of the tissue with $\alpha$-difluoromethylarginine, the inhibitor of putrescine biosynthesis, further enhanced the growth in the presence of IAA. Inhibitory effect of spermine can also be apparent for fusicoccin-induced elongation, but not for growth induced by a low pH. Spermine also suppressed the ethylene-enhanced growth in the presence of auxin. Using computer-based video digitizer system, the inhibitory effects of spermine on petiole growth were kinetically analyzed. Auxin-induced growth was characterized by an initial and transient growth with a highly elevated rate followed by a steady growth with a slightly reduced rate. Spermine treatment was found to shorten the duration of the initial phase of growth, and to reduce the rates of both the initial and steady growth as well. The latent period for auxin induction was not affected by spermine.

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Development of high tryptophan GM rice and its transcriptome analysis (고 함량 트립토판 생산 GM 벼 개발 및 전사체 분석)

  • Jung, Yu Jin;Nogoy, Franz Marielle;Cho, Yong-Gu;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.186-195
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    • 2015
  • Anthranilate synthase (AS) is a key enzyme in the biosynthesis of tryptophan (Trp), which is the precursor of bioactive metabolites like indole-3-acetic acid and other indole alkaloids. Alpha anthranilate synthase 2 (OsASA2) plays a critical role in the feedback inhibition of tryptophan biosynthesis. In this study, two vectors with single (F124V) and double (S126F/L530D) point mutations of the OsASA2 gene for feedback-insensitive ${\alpha}$ subunit of rice anthranilate synthase were constructed and transformed into wildtype Dongjinbyeo by Agrobacterium-mediated transformation. Transgenic single and double mutant lines were selected as a single copy using TaqMan PCR utilized nos gene probe. To select intergenic lines, the flanking sequence of RB or LB was digested with a BfaI enzyme. Four intergenic lines were selected using a flanking sequence tagged (FST) analysis. Expression in rice (Oryza sativa L.) of the transgenes resulted in the accumulation of tryptophan (Trp), indole-3-acetonitrile (IAN), and indole-3-acetic acid (IAA) in leaves and tryptophan content as a free amino acid in seeds also increased up to 30 times relative to the wildtype. Two homozygous event lines, S-TG1 and D-TG1, were selected for characterization of agronomic traits and metabolite profiling of seeds. Differentially expressed genes (DEGs), related to ion transfer and nutrient supply, were upregulated and DEGs related to co-enzymes that work as functional genes were down regulated. These results suggest that two homozygous event lines may prove effective for the breeding of crops with an increased level of free tryptophan content.

Characterization of Weizmannia ginsengihumi LGHNH from Wild-Ginseng and Anti-Aging Effects of Its Cultured Product (산삼 공생 미생물 Weizmannia ginsengihumi LGHNH의 특징 및 배양물의 항노화 효능)

  • Minjung Kwon;Hyejin Lee;So Young Lee;Mu Hyun Jin
    • Microbiology and Biotechnology Letters
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    • v.50 no.3
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    • pp.414-421
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    • 2022
  • In this study, we isolated Weizmannia ginsengihumi LGHNH (KCTC 14462BP) from 30-year-old wild Panax ginseng C.A. Meyer and elucidated the characteristics of the isolated bacterium and its industrial potential as an anti-aging material. W. ginsengihumi LGHNH was investigated to produce indole-3-acetic acid (IAA), a plant growth-promoting hormone (1.38 ㎍/ml to 2.22 ㎍/ml). We also confirmed the existence of bioconversion activity via the comparison of the ginsenoside content before and after fermentation. As for the converted minor ginsenoside, Rg2(R), Rg4, Rg6, Rg3(S), Rg3(R), Rk1, Rg5, Rh1(R), Rk3 and Rh4 are known to have high bioavailability and various skin effects. We measured mitochondrial membrane potential and ATP biosynthesis to elucidate W. ginsengihumi LGHNH cultured product (WCP) as an anti-aging material. As a result, the mitochondrial membrane potential in HaCaT cells with UVB decreased to 39.3% compared to the unirradiated group, but was recovered to 57.3% and 58.1% by 0.001% (v/v) and 0.01% (v/v) WCP, respectively. In addition, we measured mitochondrial ATP biosynthesis. It decreased to 94.3% compared to the unirradiated group with UVB, but was recovered to 105.3% and 105.7% by 0.001% (v/v) and 0.01% (v/v) WCP.

The Effect of Sodium Tungstate on the Aldehyde Oxidase and the Growth in the Primary Root of Maize (Zea mays) (옥수수 (Zea mays) 뿌리의 알데히드 산화효소와 생장에 미치는 텅스텐산 나트륨의 영향)

  • Oh, Young-Joo;Cho, Young-Jun;Park, Woong-June
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.990-995
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    • 2007
  • We tested the effect of sodium tungstate, which disturbs the molybdenum cofactor formation, on the activities of aldehyde oxidase(AO) and the growth of maize(Zea mays) primary roots. As reported in other plants, sodium tungstate inhibited AO also in the maize root concentration-dependently. The inhibitory effect of sodium tungstate was observed only when the inhibitor was applied to the living plants. Application of tungstate to the extracted protein did not show any effect. Western analysis revealed slightly decreased level of AO protein in the presence of tungstate, indicating a positive feedback of gene regulation by the product. We also tested the effects of tungstate on the root growth. The elongation of primary root and the development of lateral roots, which are sensitive to the absolute level of auxin, were decreased in the presence of sodium tungstate. However, the gravitropic curvature of the primary root, which is dependent on the relative amount of auxin at both sides, was unaffected. These data suggested the decrease of auxin biosynthesis by the application of tungstate. However, the level of free IAA was unaffected by tungstate application. We discuss the possible explanations for the observed results.

Effects of Methyl Jasmonate on Ethylene Producton in Tomato (Lycopersicon esculentum Mill.) Hypocotyl Segments and Fruits (Methyl jasmonate가 토마토(Lycopersicon esculentum Mill.)하배축 절편과 열매에서 에틸렌 생성에 미치는 영향)

  • June Seung Lee
    • Journal of Plant Biology
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    • v.38 no.3
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    • pp.235-242
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    • 1995
  • Effects of methyl jasmonate (MeJA) on ethylene production in tomato(Lycopersicon esculentum Mill.) hypocotyl segments and fruits were studied. Ethylene production in tomato hypocotyl segments was inhibited by the increasing concentratons of MeJA, and 450 $\mu$M of MeJA showed 50% inhibitory effect. Time course data indicate that this inhibitory effect of MeJA appeared after 3 h of incubation period and continued until 24 h. Inhibition of ethylene producton by MeJA was due to the decrease in 1-aminocyclopropane-1-carboxylic acid(ACC) synthase activity. However, MeJA treatment had no effect on ACC oxidase activity and the accumulaton of ACC oxidase mRNAs. MeJA also inhibited auxin-induced ethylene production by decreasing in ACC synthase activity. In contrast, MeJA stimulated ethylene production in tomato fruits. When 30 $\mu$L/mL MeJA was treated in a gaseous state, ethylene production doubled and this stimulating effect continued until 4 days. To investigate the mechanisms of MeJA on ethylene production, ACC synthase and ACC oxidase activities were examined after MeJA treatment. MeJA increased the activities of both ACC synthase and ACC oxidase, and induced ACC oxidase mRNA accumulation. These data suggest that MeJA plays distinct roles in the ethylene production in different tomato tissues. It is possible that MeJA affects differently the mechanisms of signal transuction leading to the ethylene biosynthesis.

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The Effect of Oligosaccharides on Ethylene Production in Mung Bean (Vigna radiata W.) Hypocotyl Segments

  • Choy, Yoon-Hi;Lee, Dong-Hee;Lee, June-Seung
    • Journal of Plant Biology
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    • v.39 no.4
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    • pp.295-300
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    • 1996
  • The physiological effects of oligogalacturonic acid (OGA:D. P. 6-7), a product of acid hydrolysis of polygalacturonic acid (PGA), on ethylene biosynthesis in mung bean (Vigna radiata W.) hypocotyl segments was studied. Among PGA, OGA and monogalacturomic acid (MGA), only OGA stimulated ethylene production in mung bean hypocotyl segments, and the most effective concentraton of OGA was 50$\mu\textrm{g}$/mL. Time course data indicated that this stimulatiion effect of OGA appeared after 90 min incubation period and continued until 24 h. When indol-3-acetic acid (IAA) and 1-aminocyclopropane-1-carboxylic acid (ACC) were treated with OGA to investigate the mechanism of OGA on ethylene production, they did not show synergistic effects on ethylene production. The stimulation of ethylene production by OGA was due to the increase of in vivo ACC synthase activity, but OGA treatment had no effect of in vivo ACC oxidase activity. The effect of aminoethoxy vinyl glycine (AVG) and Co2+, the inhibitor of ethylene synthesis, was siminished a little by the OGA, but the treatment of Ca2+, known to increase ACC, with OGA did not increase the ethylene production, this effect seems to be specific for Ca2+ because other divalent cation, Mg2+, did not show the inhibition of OGA-indyuced ethylene production. It is possible that the OGA adopts a different signal transduction pathway to the ethylene bioxynthesis.

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Enterococcus faecium LKE12 Cell-Free Extract Accelerates Host Plant Growth via Gibberellin and Indole-3-Acetic Acid Secretion

  • Lee, Ko-Eun;Radhakrishnan, Ramalingam;Kang, Sang-Mo;You, Young-Hyun;Joo, Gil-Jae;Lee, In-Jung;Ko, Jae-Hwan;Kim, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1467-1475
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    • 2015
  • The use of microbial extracts containing plant hormones is a promising technique to improve crop growth. Little is known about the effect of bacterial cell-free extracts on plant growth promotion. This study, based on phytohormonal analyses, aimed at exploring the potential mechanisms by which Enterococcus faecium LKE12 enhances plant growth in oriental melon. A bacterial strain, LKE12, was isolated from soil, and further identified as E. faecium by 16S rDNA sequencing and phylogenetic analysis. The plant growth-promoting ability of an LKE12 bacterial culture was tested in a gibberellin (GA)-deficient rice dwarf mutant (waito-C) and a normal GA biosynthesis rice cultivar (Hwayongbyeo). E. faecium LKE12 significantly improved the length and biomass of rice shoots in both normal and dwarf cultivars through the secretion of an array of gibberellins (GA1, GA3, GA7, GA8, GA9, GA12, GA19, GA20, GA24, and GA53), as well as indole-3-acetic acid (IAA). To the best of our knowledge, this is the first study indicating that E. faecium can produce GAs. Increases in shoot and root lengths, plant fresh weight, and chlorophyll content promoted by E. faecium LKE12 and its cell-free extract inoculated in oriental melon plants revealed a favorable interaction of E. faecium LKE12 with plants. Higher plant growth rates and nutrient contents of magnesium, calcium, sodium, iron, manganese, silicon, zinc, and nitrogen were found in cell-free extract-treated plants than in control plants. The results of the current study suggest that E. faecium LKE12 promotes plant growth by producing GAs and IAA; interestingly, the exogenous application of its cell-free culture extract can be a potential strategy to accelerate plant growth.