• Title/Summary/Keyword: Arabidopsis root

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Overexpression in Arabidopsis of a Plasma Membrane-targeting Glutamate Receptor from Small Radish Increases Glutamate-mediated Ca2+ Influx and Delays Fungal Infection

  • Kang, Seock;Kim, Ho Bang;Lee, Hyoungseok;Choi, Jin Young;Heu, Sunggi;Oh, Chang Jae;Kwon, Soon Il;An, Chung Sun
    • Molecules and Cells
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    • v.21 no.3
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    • pp.418-427
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    • 2006
  • Ionotropic glutamate receptors (iGluRs) are ligand-gated nonselective cation channels that mediate fast excitatory neurotransmission. Although homologues of the iGluRs have been identified in higher plants, their roles are largely unknown. In this work we isolated a full-length cDNA clone (RsGluR) encoding a putative glutamate receptor from small radish. An RsGluR:mGFP fusion protein was localized to the plasma membrane. In Arabidopsis thaliana overexpressing the fulllength cDNA, glutamate treatment triggered greater $Ca^{2+}$ influx in the root cells of transgenic seedlings than in those of the wild type. Transgenic plants exhibited multiple morphological changes such as necrosis at their tips and the margins of developing leaves, dwarf stature with multiple secondary inflorescences, and retarded growth, as previously observed in transgenic Arabidopsis overexpressing AtGluR3.2 [Kim et al. (2001)]. Microarray analysis showed that jasmonic acid (JA)-responsive genes including defensins and JA-biosynthetic genes were up-regulated. RsGluR overexpression also inhibited growth of a necrotic fungal pathogen Botrytis cinerea possibly due to up-regulation of the defensins. Based on these results, we suggest that RsGluR is a glutamate-gated $Ca^{2+}$ channel located in the plasma membrane of higher plants and plays a direct or indirect role in defense against pathogen infection by triggering JA biosynthesis.

Overexpression of ginseng patatin-related phospholipase pPLAIIIβ alters the polarity of cell growth and decreases lignin content in Arabidopsis

  • Jang, Jin Hoon;Lee, Ok Ran
    • Journal of Ginseng Research
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    • v.44 no.2
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    • pp.321-331
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    • 2020
  • Background: The patatin-related phospholipase AIII family (pPLAIIIs) genes alter cell elongation and cell wall composition in Arabidopsis and rice plant, suggesting diverse commercial purposes of the economically important medicinal ginseng plant. Herein, we show the functional characterization of a ginseng pPLAIII gene for the first time and discuss its potential applications. Methods: pPLAIIIs were identified from ginseng expressed sequence tag clones and further confirmed by search against ginseng database and polymerase chain reaction. A clone showing the highest homology with pPLAIIIβ was shown to be overexpressed in Arabidopsis using Agrobacterium. Quantitative polymerase chain reaction was performed to analyze ginseng pPLAIIIβ expression. Phenotypes were observed using a low-vacuum scanning electron microscope. Lignin was stained using phloroglucinol and quantified using acetyl bromide. Results: The PgpPLAIIIβ transcripts were observed in all organs of 2-year-old ginseng. Overexpression of ginseng pPLAIIIβ (PgpPLAIIIβ-OE) in Arabidopsis resulted in small and stunted plants. It shortened the trichomes and decreased trichome number, indicating defects in cell polarity. Furthermore, OE lines exhibited enlarged seeds with less number per silique. The YUCCA9 gene was downregulated in the OE lines, which is reported to be associated with lignification. Accordingly, lignin was stained less in the OE lines, and the expression of two transcription factors related to lignin biosynthesis was also decreased significantly. Conclusion: Overexpression of pPLAIIIβ retarded cell elongation in all the tested organs except seeds, which were longer and thicker than those of the controls. Shorter root length is related to auxinresponsive genes, and its stunted phenotype showed decreased lignin content.

Water relations of plants under environmental stresses: role of aquaporins

  • Kang, H.S.;Ahn, S.J.;Hong, S.W.;Chung, G.C.
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.71-80
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    • 2005
  • Effects of low temperature ($8^{\circ}C$) on the hydraulic conductivity of young roots of a chilling-sensitive (cucumber; Cucumis sativus L.) and a chilling-resistant (figleaf gourd; Cucurbita ficifolia Bouche) crop have been measured at the levels of whole root systems (root hydraulic conductivity, $Lp_r$) and of individual cortical cells (cell hydraulic conductivity, Lp). In figleaf gourd, there was a reduction only in hydrostatic $Lp_r$ but not in osmotic $Lp_r$ suggesting that the activity of water channels was not much affected by low root temperature (LRT)treatment in this species. Changes in cell Lp in response to chilling and recovery were similar asroot level, although they were more intense at the root level. Roots of figleaf gourd recovered better from LRT treatment than those of cucumber. In figleaf gourd, recovery (both at the root and cell level) often resulted in Lp and $Lp_r$ values which were even bigger than the original, i.e. there was an overshoot in hydraulic conductivity. These effects were larger forosmotic (representing the cell-to-cell passage of water) than for hydrostatic $Lp_r$. After a short term (1 d) exposure to $8\;^{\circ}C$ followed by 1 d at $20\;^{\circ}C$, hydrostatic $Lp_r$ of cucumber nearly recovered and that of figleaf gourd still remained higher due to the overshoot. On the contrary, osmotic $Lp_r$ and cell Lp in both species remained high by a factor of 3 as compared to the control, possibly due to an increased activity of water channels. After pre-conditioning of roots at LRT, increased hydraulic conductivitywas completely inhibited by $HgCl_2$ at both the root and cell levels. Different from figleaf gourd, recovery from chilling was not complete in cucumber after longer exposure to LRT. It is concluded that at LRT, both changes in the activity of aquaporins and alterations of root anatomy determine the water uptake in both species. To better understand the aquaporin function in plants under various stress conditions, we examined the transgenic Arabidopsisand tobacco plants that constitutively overexpress ArabidopsisPIP1;4 or PIP2;5 under various abiotic stress conditions. No significant differences in growth rates were found between the transgenic and wild-type plants under favorable growth conditions. By contrast, overexpression of PIP1;4 or PIP2;5 had a negative effect on seed germination and seedling growth under drought stress, whereas it had a positive effect under cold stress and no effect under salt stress. Measurement of water transport by cell pressure probe revealed that these observed phenotypes under different stress conditions were closely correlated with the ability of water transport by each aquaporin in the transgenic plants. Together, our results demonstrate that PIP-type aquaporins play roles in seed germination, seedling growth, and stress response of Arabidopsis and tobacco plants under various stress conditions, and emphasize the importance of a single aquaporin-mediated water transport in these cellular processes.

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Effect of Ethylene on Formations of Adventitious Roots, Trichomes, and Calli by NAA in Leaf segments of Arabidopsis thaliana (애기장대 잎 절편 배양시 NAA 농도에 따른 부정근, 모용 및 캘러스 형성에 미치는 에틸렌의 영향)

  • 한태진;홍종필;김준철;임창진;진창덕
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.259-264
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    • 1999
  • In order to study the role of ethylene on the formation of adventitious roots, trichomes and calli, the effects of 1-aminocyclopropane-l-carboxylic acid (ACC), ethephon, $CoCl_2$ and $AgNO_3$ were investigated in the leaf segments from ecotype Columbia of Arabidopsis thaliana. When the leaf segments were cultured on the media for forming adventitious roots (0.1 mg/L NAA), trichomes (2.0 mg/L NAA) and calli (10.0 mg/L NAA), and then each cultures was treated with 1-100 mg/L of ACC and ethephon, respectively. On the adventitious root-forming medium adventitious root formation was decreased, and trichomes were induced. And on the trichome-forming medium trichome formation was decreased, and calli were induced. In order hand each culture was treated with 1-100 mg/L of $CoCl_2$ and $AgNO_3$, respectively. On the adventitious root-forming medium adventitious roots was increased without trichome formation, and on the trichome-forming medium trichome formation was decreased, and adventitious roots were induced. However on the callus-forming medium treated with ACC, ethephon, $CoCl_2$ and $AgNO_3$, respectively, callus formation was inhibited and trichomes were induced in all cultures.

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Cloning and Characterization of a Rice cDNA Encoding Glutamate Decarboxylase

  • Oh, Suk-Heung;Choi, Won-Gyu;Lee, In-Tae;Yun, Song-Joong
    • BMB Reports
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    • v.38 no.5
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    • pp.595-601
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    • 2005
  • In this study, we have isolated a rice (Oryza sativa L.) glutamate decarboxylase (RicGAD) clone from a root cDNA library, using a partial Arabidopsis thaliana GAD gene as a probe. The rice root cDNA library was constructed with mRNA, which had been derived from the roots of rice seedlings subjected to phosphorus deprivation. Nucleotide sequence analysis indicated that the RicGAD clone was 1,712 bp long, and harbors a complete open reading frame of 505 amino acids. The 505 amino acid sequence deduced from this RicGAD clone exhibited 67.7% and 61.9% identity with OsGAD1 (AB056060) and OsGAD2 (AB056061) in the database, respectively. The 505 amino acid sequence also exhibited 62.9, 64.1, and 64.2% identity to Arabidopsis GAD (U9937), Nicotiana tabacum GAD (AF020425), and Petunia hybrida GAD (L16797), respectively. The RicGAD was found to possess a highly conserved tryptophan residue, but lacks the lysine cluster at the C-proximal position, as well as other stretches of positively charged residues. The GAD sequence was expressed heterologously using the high copy number plasmid, pVUCH. Our activation analysis revealed that the maximal activation of the RicGAD occurred in the presence of both $Ca^{2+}$ and calmodulin. The GAD-encoded 56~58 kDa protein was identified via Western blot analysis, using an anti-GAD monoclonal antibody. The results of our RT-PCR analyses revealed that RicGAD is expressed predominantly in rice roots obtained from rice seedlings grown under phosphorus deprivation conditions, and in non-germinated brown rice, which is known to have a limited phosphorus bioavailability. These results indicate that RicGAD is a $Ca^{2+}$/calmodulin-dependent enzyme, and that RicGAD is expressed primarily under phosphate deprivation conditions.