• Title/Summary/Keyword: Arabidopsis mutant

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Protoplast Isolation and Regeneration of Fertile Plants from Arabidopsis Trp Mutant, trp1-100

  • Lim, Seon-hee;Kim, Young-soon;Lee, Eui-seung;Rose, Alan;Last, Robert;Cheong, Hyeon-sook
    • Animal cells and systems
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    • v.2 no.2
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    • pp.239-242
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    • 1998
  • Arabidopsis trp1 mutant plants, deficient in phosphoribosyI anthranilate transferase (PAT) activity, accumulate anthranilate compounds, which render them blue fluorescence. The visible phenotype of trp1 makes the PAT gene an excellent reporter gene in the mutant. In order to develop a system for the homologous recombination using the phenotypic characteristic of trp1-100, we established optimum conditions for the isolation and regenera tion of protoplast from auxin-conditioned, trp1-100 root cultures. Trvptophan had to be supplemented in the germination medium for the efficient cell division and subsequent plant regeneration. When 10 uM tryptophan was added to the germination medium, we obtained the highest yield of protoplasts ($3{\times}10^6 cells/g$) and the best viability (92%). Thirty percent of root protoplast derived from meristematic cells underwent cell division within 5 days in callus-induction medium. Regenerated rosette leaves (2-3 mm) were transferred to rooting medium and finally acclimated to the soil for flowering.

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Effect of Light on Root Growth and Gravitropic Response of Phytochrome Mutants of Arabidopsis (Arabidopsis phytochrome mutant에서 빛이 뿌리 생장과 굴중성 반응에 미치는 영향)

  • Park, Ji-Hye;Lee, Sang-Seoung;Woo, Soon-Hwa;Kim, Soon-Young
    • Journal of Life Science
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    • v.22 no.5
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    • pp.681-686
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    • 2012
  • Light, one of the environmental stimuli, is fundamental to the growth and development of plants. Red and far-red light are sensed using the phytochrome family of plant photoreceptors. To investigate the effect of light on root growth and gravitropism, we used the Arabidopsis phytochrome mutants grown in several light conditions. The root growth of $phyA$ reared in all light conditions except white light and was stimulated compared to the WT. The stimulation of root growth was obvious in $phyA$ grown in red light. On the other hand, the root growth of $phyB$ grown in all light conditions decreased, and the lowest rate of decrease was observed in $phyAB$ grown in white and red light. The gravitropic response of $phyA$ was stimulated compared to the WT when it was grown in all light conditions except far-red light. $PhyAB$ grown in all light conditions showed the inhibition of gravitropic response. The transcript level of ACS, one of the enzymes regulating ethylene biosynthesis, increased in $phyA$ grown in white and red light, but not in $phyA$ grown in far-red light. In conclusion, these results suggested that the $P_{fr}$ form of $phyB$ regulates the root growth and gravitropism.

Regulation of cell size and cell number by LANCEOLATA1 gene in Arabidopsis (애기장대의 세포 크기와 세포 수를 조절하는 LANCEOLATA1 유전자)

  • Cho, Kiu-Hyung;Jun, Sang-Eun;Jeong, Soon-Jae;Yi, Young-Byung;Kim, Gyung-Tae
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.1-5
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    • 2007
  • The Precesses for leaf development in dicotyledonous plants are surprisingly complex, while the mechanism of controlling and coordinating them is poorly understood. To characterize the fundamental features of the leaf development of Arabidopsis, we first attempted to isolate mutants that alter leaf morphology. Here, leaf morphological mutant of Arabidopsis, lanceolatal (lan1) which has small and narrow leaves have isolated and characterized. To clarify the function of LAN1 in organ development, we characterized lan1-7 mutant using an anatomical and genetic approach. The lan1-7 mutant had reduced size of foliage leaves and reduced dimensions of stems. A reduction both in cell size and in cell number was evident at the cellular level in the lan1 mutant, revealing that LAN1 gene appears to affect cell division at an earlier stage and cell elongation throughout the development of leaf primordia. from the analysis of heterogeneous plant with lan1 mutation and 35S-AG transgenic plant, AG gene is revealed to regulate leaf morphology under the control of 35S promoter. Thus, MADS-box gene was revealed to have some relationship to that of LAN1 gene at certain stage in leaf development processes.

Functional analysis of the rice BRI1 receptor kinase (벼 Brassinosteroid Insensitive 1 Receptor Kinase의 기능에 관한 연구)

  • Yeon, Jinouk;Kim, Hoy-Taek;Nou, Ill-Sup;Oh, Man-Ho
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.30-36
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    • 2016
  • Brassinosteroids (BRs) are essential plant steroid hormones required for cell elongation, plant growth, development and abiotic and biotic stress tolerance. BRs are recognized by BRI1 receptor kinase that is localized in the plasma membrane, and the BRI1 protein will eventually autophosphorylate in the intracellular domain and transphosphorylate BAK1, which is a co-receptor in Arabidopsis thaliana. However, little is known of the role OsBRI1 receptor kinase plays in Oryza sativa, monocotyledonous plants, compared to that in Arabidopsis thaliana, dicotyledonous plants. As such, we have studied OsBRI1 receptor kinase in vitro and in vivo with recombinant protein and transgenic plants, whose phenotypes were also investigated. A OsBRI1 cytoplasmic domain (CD) recombinant protein was induced in BL21 (DE3) E.coli cells with IPTG, and purified to obtain OsBRI1 recombinant protein. Based on Western blot analysis with phospho-specific pTyr and pThr antibodies, OsBRI1 recombinant protein and OsBRI1-Flag protein were phosphorylated on Threonine residue(s), however, not on Tyrosine residue(s), both in vitro and in vivo. This is particularly intriguing as AtBRI1 protein was phosphorylated on both Ser/Thr and Tyr residues. Also, the OsBRI1 full-length gene was expressed in, and rescued, bri1-5 mutants, such as is seen in normal wild-type plants where AtBRI1-Flag rescues bri1-5 mutant plants. Root growth in seedlings decreased in Ws2, AtBRI1, and 3 independent OsBRI1 transgenic seedlings and had an almost complete lack of response to brassinolide in the bri1-5 mutant. In conclusion, OsBRI1, an orthologous gene of AtBRI1, can mediate normal BR signaling for plant growth and development in Arabidopsis thaliana.

The Expression Patterns of AtBSMT1 and AtSAGT1 Encoding a Salicylic Acid (SA) Methyltransferase and a SA Glucosyltransferase, Respectively, in Arabidopsis Plants with Altered Defense Responses

  • Song, Jong Tae;Koo, Yeon Jong;Park, Jong-Beum;Seo, Yean Joo;Cho, Yeon-Jeong;Seo, Hak Soo;Choi, Yang Do
    • Molecules and Cells
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    • v.28 no.2
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    • pp.105-109
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    • 2009
  • We reported previously that overexpression of a salicylic acid (SA) methyltransferase1 gene from rice (OsBSMT1) or a SA glucosyltransferase1 gene from Arabidopsis thaliana (AtSAGT1) leads to increased susceptibility to Pseudomonas syringae due to reduced SA levels. To further examine their roles in the defense responses, we assayed the transcript levels of AtBSMT1 or AtSAGT1 in plants with altered levels of SA and/or other defense components. These data showed that AtSAGT1 expression is regulated partially by SA, or nonexpressor of pathogenesis related protein1, whereas AtBSMT1 expression was induced in SA-deficient mutant plants. In addition, we produced the transgenic Arabidopsis plants with RNAi-mediated inhibition of AtSAGT1 and isolated a null mutant of AtBSMT1, and then analyzed their phenotypes. A T-DNA insertion mutation in the AtBSMT1 resulted in reduced methyl salicylate (MeSA) levels upon P. syringae infection. However, accumulation of SA and glucosyl SA was similar in both the atbsmt1 and wild-type plants, indicating the presence of another SA methyltransferase or an alternative pathway for MeSA production. The AtSAGT1-RNAi line exhibited no altered phenotypes upon pathogen infection, compared to wild-type plants, suggesting that (an)other SA glucosyltransferase(s) in Arabidopsis plants may be important for the pathogenesis of P. syringae.

Molecular cloning of the Arabidopsis gene rat3 that is involving in the Agobacterium-mediated planttransformation (Agrobacterium에 의한 식물형질전환에 관여하는 Arabidopsis RAT3 유전자의 분리와 분석)

  • 남재성;양보경;김도훈;정순재;이영병
    • Journal of Life Science
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    • v.11 no.5
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    • pp.423-431
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    • 2001
  • Genomic and cDNA clones containing the RAT3 gene involving in Agobacterium-mediated plant transformation were identified using plant DNA flanking the righ border of a T-DNA rescued from the rat3 mutant as hy-bridization probe. Two highly homologous cDNA clones were identified; one (RAT3-1) weakly hybridized with the probe whereas another (RAT3-2) strongly hybridized with the probe. Both Rat3-1 and Rat3-2 proteins contain a putative signal peptide for secretion. The deduced molecular weights of encoded proteins are 15 kDa. The results of genomic DNA blot analysis and DNA sequencing indicated that RAT3-1 and RAT3-2 exist as single copy genes and they were arranged side by side with just 600 bp distance between them. RAT3-1 was disrupted by the integration of T-DNA into the 3 untranslated region in rat3 mutant. A BLAST search showed that both RAT3-1 and RAT3-2 proteins have homology with only the C-terminal region of $\beta$-1,3-glucanase homologues from Triticum aestivum and Arabidopsis thaliana. Thses $\beta$-1,3-glucanase homologues contain an unusually long C-terminal region with no sig-nificant homology to other $\beta$-1,3-glucanase.

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The vacuolar processing enzyme (VPE) mutation suppresses an HR-like cell death induced by the double knockout mutant of vacuolar Ca2+-ATPases in Arabidopsis (애기장대에서 두 액포막 칼슘펌프 돌연변이에 의하여 유도되는 세포사멸 표현형의 액포수식효소(VPE) 돌연변이에 의한 억제)

  • Park, Hyeong-Cheol;Lee, Sang-Min;Kim, Ho-Soo;Chung, Woo-Sik
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.169-175
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    • 2011
  • Calcium ($Ca^{2+}$) signals have been implicated in regulating plant development and responses to the environmental stresses including a programmed cell death pathway. In animals and plants, cytosolic $Ca^{2+}$ signals have been involved in the activation of programmed cell death (PCD). Recently, we reported that disruption of Arabidopsis vacuolar $\b{A}$utoinhibited $\underline{C}a^{2+}$-$\b{A}$TPases (ACAs), ACA4 and ACA11, resulted in the activation of a salicylic acid-dependent programmed cell death pathway. Although extensive studies have revealed various components of a PCD in plants, executors to directly induce PCD are well unknown. Here, we provide that the vacuolar processing enzymes (VPEs) are involved in a PCD induced by the double knockout mutant of vacuolar $Ca^{2+}$-ATPases in Arabidopsis. The gene expression of VPE was rapidly up-regulated and the enzyme activity of VPE was increased in the double mutant plants. We also generated aca4/aca11/avpe, aca4/aca11/${\gamma}$vpe and aca4/aca11/avpe/${\gamma}$vpe mutant plants. Although cell death phenotype of the double mutant plants was not completely disappeared in the triple and quadruple mutant plants, the triple and quadruple mutant plants showed to significantly delay cell death phenotype of the double mutant plants. These results suggest that the VPE is involved in the HR-like cell death in the double mutant of vacuolar $Ca^{2+}$-ATPases in Arabidopsis.

Identification of Plant Factors Involving in Agrobacterium-mediated Plant Transformation

  • Nam, Jaesung
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.387-393
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    • 2000
  • The process by which Agrobacterium tumefaciens genetically transforms plants involves a complex series of reactions communicated between the pathogen and the plants. To identify plant factors involved in agrobacterium-mediated plant transformation, a large number of T-DNA inserted Arabidopsis thaliana mutant lines were investigated for susceptibility to Agrobacterium infection by using an in vitro root inoculation assay. Based on the phenotype of tumorigenesis, twelve T-DNA inserted Arabidopsis mutants(rat) that were resistant to Agrobacterium transformation were found. Three mutants, rat1, rat3, and rat4 were characterized in detail. They showed low transient GUS activity and very low stable transformation efficiency compared to the wild-type plant. The resistance phenotype of rat1 and rats resulted from decreased attachment of Agrobacterium tumefaciens to inoculated root explants. They may be deficient in plant actors that are necessary for bacterial attachment to plant cells. The disrupted genes in rat1, rat3, and rat4 mutants were coding a arabinogalactan protein, a likely cell wall protein and a cellulose synthase-like protein, respectively.

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Geness for degradation of storage oil and their application to oil biotechnology

  • Nishimura, Mikio;Hayashi, Makoto;Kato, Akira;Mano, Shoji;Hayashi, Hiroshi;Yamaguchi, Katushi;Nito, Kazumasa;Fukao, Youichiro
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.37-40
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    • 1999
  • cDNAs for long- and short-chain acyl-CoA oxidases in fatty acid $\beta$-oxidation were isolated and were characterized their enzymatical and molecular properties. Both oxidases were exclusively localized in glyoxysomes, indicating that glyoxysomes can completely metabolize fatty acids to acyl-CoA by their cooperative action. In order to clarify the regulatory mechanisms underlying degradation of storage oil, we tried to obtain glyoxysome-deficient mutants of Arabidopsis. We screened 2,4-dichlorophenoxybutyric acid (2,4-DB) mutants of Arabidopsis which have defects in glyoxysomal fatty acid $\beta$-oxidation. Four mutants can be classified as carrying alleles at three independent loci, which we designated pedl, ped2, and ped3, respectively (where ped stands for peroxisome defective). The characteristics of these ped mutants are described.

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Auxin Induced Expression of Expansin is Alered in a New Aux1 Allele that Shows Severe Defect in Gravitropic Response

  • Jeong, Hae-Jun;Kwon, Ye-Rim;Oh, Jee-Eun;Kim, Ki-Deok;Lee, Sung-Joon;Hong, Suk-Whan;Lee, Ho-Joung
    • Journal of Applied Biological Chemistry
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    • v.49 no.4
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    • pp.148-153
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    • 2006
  • While the underlying molecular mechanism remains to be elucidated, recent studies suggest that polar auxin transport is a key controlling factor in triggering differential growth responses to gravity. Identification of regulatory components in auxin-mediated differential cell expansion would improve our understanding of the gravitropic response. In this study, we identify a mutant designated aux1-like(later changed to aux1), an allele of the aux1 mutant that exhibits a severely disrupted root gravitropic response, but no defects in developmental processes. In Arabidopsis, AUX1 encodes an auxin influx carrier. Since in-depth characterization of the gravitropic response caused by mutations in this gene has been performed previously, we focused on identifying the downstream genes that were differentially expressed compared to wild-type plants. Consistent with the mutant phenotype, the transcription of the auxin-responsive genes IAA17 and GH3 were altered in aux1 plants treated with IAA, 2, 4-D and NAA. In addition, we identified two expansin genes EXP10 and EXPL3 that exhibited different expression in wild-type and mutant plants.