• Title/Summary/Keyword: Arabidopsis mutant

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Design and Implementation of 08MS-based Plant Mutant Proteome Analysis System (DBMS 기반 식물 돌연변이 단백질체 분석을 위한 시스템 설계 및 구현)

  • 허정호;박춘구;박윤주;최지인;정동수;남홍길
    • Proceedings of the Korean Information Science Society Conference
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    • 2002.10c
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    • pp.283-285
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    • 2002
  • 생물학의 주된 목적이 세포 안에서의 유전자의 기능을 알아내는 것이므로 유전자의 실제 기능을 담당하는 단백질에 대한 연구는 필연적인 것이다. 근래에 이르러 2DE등의 실험방법 개선 둥으로 대량의 단백질에 대한 연구가 실현화됨에 따라, 프로트오믹스(proteomics)의 연구가 활성화되었으며, 본 실험팀 역시 식물 애기장대의 돌연변이들을 이용한 프로티오믹스 연구를 진행중이다. 프로티오믹스 연구는 실험 특성상 대량의 단백질 정보들을 얻게 되므로 데이터의 수작업 분석이 불가능하다. 그리하여 우리는 식물 애기장대(arabidopsis)의 특성과 2DE 실험에서 파생되는 여러 데이터들을 반영하여 정보를 효율적으로 관리할 수 있는 데이터 베이스를 설계 구축하였으며, 나아가 각 표현형을 대표할 수 있는 표지 단백질을 선별해내는 분석모듈을 구현하였다.

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Isolation of SYP61/OSMl that is Required for Salt Tolerance in Arabidopsis by T-DNA Tagging (애기장대에서 고염 스트레스 내성에 관여하는 OSM1/SYP61 유전자의 동정)

  • Kim, Ji-Yeon;Baek, Dong-Won;Lee, Hyo-Jung;Shin, Dong-Jin;Lee, Ji-Young;Choi, Won-Kyun;Kim, Dong-Giun;Chung, Woo-Sik;Kwak, Sang-Soo;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.11-18
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    • 2006
  • Salt stress is one of major environmental factors influencing plant growth and development. To identify salt tolerance determinants in higher plants, a large-scale screen was conducted with a bialaphos marker-based T-DNA insertional collection of Arabidopsis ecotype C24 mutants. One line for salt stress-sensitive mutant (referred to as ssm1) exhibited increased sensitivity to both ionic (NaCl) and nonionic (mannitol) osmotic stress in a root growth assay. This result suggests that ssm1 mutant is involved in ion homeostasis and osmotic compensation in plant. Molecular cloning of the genomic DNA flanking T-DNA insert of ssm1 mutant was achieved by mutant genomic DNA library screening. T-DNA insertion appeared in the first exon of an open reading frame on F3M18.7, which is the same as AtSYP61. SSM1 is SYP61/OSM1 that is a member of the SNARE superfamily of proteins required for vesicular/target membrane fusions and factor related to abiotic stress.

Functional characterization of Arabidopsis thaliana BLH 8, BEL1-Like Homeodomain 8 involved in environmental stresses (환경 스트레스에 관여하는 애기장대 BLH 8, BEL1-Like Homeodomain 8의 기능 분석)

  • Park, Hyeong-Cheol;Park, Ji-Young;Baek, Dong-Won;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.162-168
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    • 2011
  • High salinity is a common stress condition that adversely affects plant growth and crop production. In response to various environmental stresses, plants activate a number of defense genes that function to increase the tolerance. To isolate Arabidopsis genes that are involved in abiotic stress responses, we carried out genetic screening using various mutant lines. Among them, the blh8 ($\b{B}$EL1-$\b{L}$ike $\b{H}$omeodomain $\underline{8}$) mutant specifically shows chlorotic phenotypes to ionic (specifically, $Na^+$ and $K^+$) stresses, but no differences in root growth. In addition, BLH8 is related to plant development and abiotic stress as predicted by a Graphical Gaussian Model (GGM) network program. It implies that BLH8 functions as a putative transcription factor related to abiotic stress responses. Collectively, our results show that gene network analysis is a useful tool for isolating genes involved in stress adaptation in plants.

Isolation and characterization of induced disease resistance (ISR)-deficient mutants of a biocontrol bacterium Pseudomonas chlororaphis O6.

  • Han, Song-Hee;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.101.1-101
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    • 2003
  • Lipopolysaccharide, siderophore, and cyclic dipeptide have been shown to be necessary for ISR induction by pseudomnads. However, there is no report on cloning of genes or generating specific mutants involving in ISR activity. A biological control bacteium P. chlororaphis O6 induces resistance to Erwinia carotovora subsp. carotovara SCCI in tobacco and induces drought resistance in Arabidopsis. To isolate genes involved in ISR activity and induction of drough resistance of O6, we constructed Tn5 mutants and were used to screen for ISR activity and drought resistance activity using microtiter assay with tobacco and Arabidopsis. Thirty-three ISR-deficient mutants were selected, and the nine ISR-deficient mutants were also lost activity of drought resistance. The flanking sequence analysis of the ISR and drought resistance-deficient mutants showed that a gacS gene encoding a two-component sensor kinase, and a mce gene encoding a protein involved in mycobacterial cell entry were mutated. The flanking sequence of each Tn5 mutant altered ISR activity is currently under investigation. These results indicate that gacS and mce are important genes in induction of ISR activity and drought resistance of P. chlororaphis O6. Our works will open opportunities for identification of bacterial genes or traits that are involved in ISR activity and induced drought resistance of P. chlororaphis O6.

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Molecular Genetic Analysis of Leaf Senescence in Arabidopsis

  • Woo, Hye-Ryun;Lee, Ung;Cho, Sung-Whan;Lim, Pyung-Ok;Nam, Hong-Gil
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.4
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    • pp.259-268
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    • 2000
  • Senescence is a sequence of biochemical and physiological events that lead to death of a cell, organ, or whole organism. Senescence is now clearly regarded as a genetically determined and evolutionarilly acquired developmental process comprising the final stage of development. However, in spite of the biological and practical importance, genetic mechanism of senescence has been very limited. Through forward and reverse genetic approaches, we are trying to reveal the molecular and genetic mechanism of senescence in plants, employing leaf organs of Arabidopsis as a model system. Using forward genetic approach, we have initially isolated several delayed senescence mutants either from T-DNA insertional lines or chemical-mutagenized lines. In the case of ore 4 and ore 9 mutants, the mutated genes were identified. The recent progress on characterization of mutants and identification of the mutated genes will be reported. We are also screening mutations from other various sources of mutant pools, such as activation tagging lines and promoter trap lines. Two dominant senescence-delayed mutants were isolated from the activation tagging pool. Cloning of the genes responsible for this phenotype is in progress. For reverse genetic approach, the genes that induced during leaf senescence were first isolated by differential screening method. We are currently using PCR-based suppression subtractive hybridization, designed to enrich a cDNA library for rare differentially expressed transcripts. Using this method, we have identified over 35 new sequences that are upregulated at leaf senescence stage. We are investigating the function of these novel genes by systemically generating antisense lines.

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Rice 7-Hydroxymethyl Chlorophyll a Reductase Is Involved in the Promotion of Chlorophyll Degradation and Modulates Cell Death Signaling

  • Piao, Weilan;Han, Su-Hyun;Sakuraba, Yasuhito;Paek, Nam-Chon
    • Molecules and Cells
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    • v.40 no.10
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    • pp.773-786
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    • 2017
  • The loss of green coloration via chlorophyll (Chl) degradation typically occurs during leaf senescence. To date, many Chl catabolic enzymes have been identified and shown to interact with light harvesting complex II to form a Chl degradation complex in senescing chloroplasts; this complex might metabolically channel phototoxic Chl catabolic intermediates to prevent oxidative damage to cells. The Chl catabolic enzyme 7-hydroxymethyl Chl a reductase (HCAR) converts 7-hydroxymethyl Chl a (7-HMC a) to Chl a. The rice (Oryza sativa) genome contains a single HCAR homolog (OsHCAR), but its exact role remains unknown. Here, we show that an oshcar knockout mutant exhibits persistent green leaves during both dark-induced and natural senescence, and accumulates 7-HMC a and pheophorbide a (Pheo a) in green leaf blades. Interestingly, both rice and Arabidopsis hcar mutants exhibit severe cell death at the vegetative stage; this cell death largely occurs in a light intensity-dependent manner. In addition, 7-HMC a treatment led to the generation of singlet oxygen ($^1O_2$) in Arabidopsis and rice protoplasts in the light. Under herbicide-induced oxidative stress conditions, leaf necrosis was more severe in hcar plants than in wild type, and HCAR-overexpressing plants were more tolerant to reactive oxygen species than wild type. Therefore, in addition to functioning in the conversion of 7-HMC a to Chl a in senescent leaves, HCAR may play a critical role in protecting plants from high light-induced damage by preventing the accumulation of 7-HMC a and Pheo a in developing and mature leaves at the vegetative stage.

Cytokinin signaling promotes root secondary growth and bud formation in Panax ginseng

  • Kyoung Rok Geem;Yookyung Lim;Jeongeui Hong;Wonsil Bae;Jinsu Lee;Soeun Han;Jinsu Gil;Hyunwoo Cho;Hojin Ryu
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.220-228
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    • 2024
  • Background: Panax ginseng, one of the valuable perennial medicinal plants, stores numerous pharmacological substrates in its storage roots. Given its perennial growth habit, organ regeneration occurs each year, and cambium stem cell activity is necessary for secondary growth and storage root formation. Cytokinin (CK) is a phytohormone involved in the maintenance of meristematic cells for the development of storage organs; however, its physiological role in storage-root secondary growth remains unknown. Methods: Exogenous CK was repeatedly applied to P. ginseng, and morphological and histological changes were observed. RNA-seq analysis was used to elucidate the transcriptional network of CK that regulates P. ginseng growth and development. The HISTIDINE KINASE 3 (PgHK3) and RESPONSE REGULATOR 2 (PgRR2) genes were cloned in P. ginseng and functionally analyzed in Arabidopsis as a two-component system involved in CK signaling. Results: Phenotypic and histological analyses showed that CK increased cambium activity and dormant axillary bud formation in P. ginseng, thus promoting storage-root secondary growth and bud formation. The evolutionarily conserved two-component signaling pathways in P. ginseng were sufficient to restore CK signaling in the Arabidopsis ahk2/3 double mutant and rescue its growth defects. Finally, RNA-seq analysis of CK-treated P. ginseng roots revealed that plant-type cell wall biogenesis-related genes are tightly connected with mitotic cell division, cytokinesis, and auxin signaling to regulate CK-mediated P. ginseng development. Conclusion: Overall, we identified the CK signaling-related two-component systems and their physiological role in P. ginseng. This scientific information has the potential to significantly improve the field-cultivation and biotechnology-based breeding of ginseng.

Cloning and Expression of a Rice cDNA Encoding a Lls1 Homologue of Maize

  • Jwa, Nam-Soo;Park, Sang-Geun;Park, Chan-Ho;Kim, Soon-Ok;Ahn, Il-Pyung;Park, Sook-Young;Yoon, Choong-Hyo;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.16 no.3
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    • pp.151-155
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    • 2000
  • A cDNA (Oslls1) encoding Lls1-homologue of maize was isolated from cDNA library of rice (Oryza sativa cv. Ilpum). The 2,138 bp of full length Oslls1 clone contains an open reading frame of 1,623 nucleotides encoding 575 amino acid residues. The deduced amino acid sequence of Oslls1 has a high level of homology with chlorophyll a oxygenases of Arabidopsis thaliana (67%) and Marchantia polymorpha (65%). Southern blot analysis of genomic DNA indicates the existence of a small gene family for Oslls1 in the rice genome. The expression of Oslls1 mRNA was induced in leaves and germinating seeds. Treatment of $H_2O$$_2$significantly down-regulated Oslls1 expression. The expression of Oslls1 mRNA was consititutively down-regulated in the blm, a rice mutant exhibiting spontaneous necrotic lesions. These results suggest that this Oslls1 gene may be involved incell death mechanisms in the blm mutant of rice.

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