• Title/Summary/Keyword: Ectopic development

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Trehalose Metabolism: Gate to Stress Signaling and Seed Development in Plant\ulcorner

  • Chung, H-J;Kim, Y-S;Lee, E-J;Kim, J-S;Shin, Y-M;Cho, I-S;Jin, H-O;Cho, J-W;Chung, C-H
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.5
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    • pp.415-421
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    • 2000
  • The disaccharide trehalose ($\alpha$-D-glucopyranosyl-$\alpha$-D-glucopyranoside) is found in variety of organ-isms that are able to withstand almost complete desiccation. In order to identify the function of trehalose in plants, we isolated Arabidopsis trehalase (AtTRE) gene that encodes the enzyme able to hydrolyze trehalose to glucose, and trehalose-6-phosphate synthase isolog, TPS3 gene by RT-PCR. The AtTRE had the substrate specificity to hydrolyze only trehalose, and a broad pH range of enzyme activity. The AtTRE promoter/GUS reporter gene was expressed in cotyledons, mature leaf tissues including guard cells, and developing siliques. The GUS expression driven by AtTPS3 promoter was significant in root tissues, and the level of GUS activity was much higher than that of the pBll 21 control seedlings. The knockout of AtTPS3 gene in Arabidopsis resulted in the retarded root development, whereas the overexpression of AtTPS3 increased the root elongation in the presence of sucrose in MS medium. Possible functions of AtTRE and AtTPS3 in plant will be discussed. In addition, ectopic expression of yeast TPS1 driven by the inducible promoters in tobacco and potato conferred the plants on the drought and freezing tolerances.

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Increased biomass and enhanced tolerance to salt stress in Chinese cabbage overexpressing Arabidopsis H+-PPase (AVP1) (애기장대 H+-PPase(AVP1) 과발현 배추에서 바이오매스 증가와 내염성 향상)

  • Park, Mehea;Won, Hee-Yeun;Kim, Chang Kil;Han, Jeung-Sul
    • Journal of Plant Biotechnology
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    • v.39 no.4
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    • pp.253-260
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    • 2012
  • On the basis of the reported agriculturally valuable phenotypes resulted from ectopic overexpression of Arabidopsis vacuolar $H^+$-PPase (AVP1), we generated the Chinese cabbage lines expressing AVP1 which then subjected to salt stress to determine the AVP1 expression if it consistently confers the capability for increasing biomass and enhancing tolerance to salinity in other species. Collectively, here we demonstrate that the transgenic young plants show more vigorous growth and higher tolerance to salt stress than wild-type ones. Increased biomass phenotype by AVP1 expression was supported by comparing fresh and dry weights of transgenic and wild type plants grown under normal condition, while higher salt tolerance trait was confirmed by tracing the kinetics of photosystem II quantum yield and DAB-staining under gradually intensified salt stress induced by MS salt or NaCl, followed by normal condition.

Actin Cytoskeleton and Golgi Involvement in Barley stripe mosaic virus Movement and Cell Wall Localization of Triple Gene Block Proteins

  • Lim, Hyoun-Sub;Lee, Mi Yeon;Moon, Jae Sun;Moon, Jung-Kyung;Yu, Yong-Man;Cho, In Sook;Bae, Hanhong;DeBoer, Matt;Ju, Hojong;Hammond, John;Jackson, Andrew O.
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.17-30
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    • 2013
  • Barley stripe mosaic virus (BSMV) induces massive actin filament thickening at the infection front of infected Nicotiana benthamiana leaves. To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB) proteins were coexpressed in cells with the actin marker DsRed: Talin. TGB ectopic expression experiments revealed that TGB3 is a major elicitor of filament thickening, that TGB2 resulted in formation of intermediate DsRed:Talin filaments, and that TGB1 alone had no obvious effects on actin filament structure. Latrunculin B (LatB) treat-ments retarded BSMV cell-to-cell movement, disrupted actin filament organization, and dramatically decreased the proportion of paired TGB3 foci appearing at the cell wall (CW). BSMV infection of transgenic plants tagged with GFP-KDEL exhibited membrane proliferation and vesicle formation that were especially evident around the nucleus. Similar membrane proliferation occurred in plants expressing TGB2 and/or TGB3, and DsRed: Talin fluorescence in these plants colocalized with the ER vesicles. TGB3 also associated with the Golgi apparatus and overlapped with cortical vesicles appearing at the cell periphery. Brefeldin A treatments disrupted Golgi and also altered vesicles at the CW, but failed to interfere with TGB CW localization. Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3.

The Panax ginseng Flowering Locus T Shows Age Specific Expression Pattern in Ginseng and Increases Root Length in Transgenic Arabidopsis

  • Mohanan, Padmanaban;Myagmarav, Davajargal;Zhang, Dabing;Kim, Yu-Jin;Yang, Deok-Chun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.17-17
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    • 2018
  • Panax ginseng Meyer is a perennial medicinal plant, the roots of which has been used in the traditional formulations in Oriental countries. To understand its floral transition, we isolated Flowering Locus T (FT) from ginseng, the bioinformatics analysis of PgFT has revealed a considerable homology to the higher plants, with the essential amino acids for FT function are conserved. The phylogenetic analysis has shown that the PgFT is belonged to the shrub classes of plants and closest kin to Jatropha curcas FT. The expression profiling from juvenile (2-year-old) were abundant in leaves as well as in root and was concentrated in the secondary leaflet and stem bottom in adult (4-year-old) ginseng plant tissues, moreover PgFT transcript displayed photoperiod dependent oscillation. The ectopic expression of PgFT in Arabidopsis thaliana, exhibit precocious flowering and several floral pathway integrators were up-regulated, interestingly their root length was increased in the transgenic seedlings. Therefore, we could conclude that PgFT encodes a florigen that acts as a key regulator in the flowering pathway in ginseng and hypothesize that, it might involve in the underground organ development as well. We believe our finding could provoke future studies on the physiology and development in P. ginseng.

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Functional Screening of Plant Genes Suppressed Salt Sensitive Phenotype of Calcineurin Deficient Mutant through Yeast Complementation Analysis (애기장대의 염해 저항성 관련 유전자의 기능적 선별)

  • Moon, Seok-Jun;Park, Soo-Kwon;Hwang, Un-Ha;Lee, Jong-Hee;Han, Sang-Ik;Nam, Min-Hee;Park, Dong-Soo;Shin, Dongjin
    • Journal of Life Science
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    • v.23 no.1
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    • pp.1-7
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    • 2013
  • Understanding salt tolerance mechanisms is important for the increase of crop yields, and so, several screening approaches were developed to identify plant genes which are involved in salt tolerance of plants. Here, we transformed the Arabidopsis cDNA library into a salt-sensitive calcineurin (CaN)-deficient ($cnb{\Delta}$) yeast mutant and isolated the colonies which can suppress salt-sensitive phenotype of $cnb{\Delta}$ mutant. Through this functional complementation screen, a total of 34 colonies functionally suppressed the salt-sensitive phenotype of $cnb{\Delta}$ yeast cells, and sequencing analysis revealed that these are 9 genes, including CaS, AtSUMO1 and AtHB-12. Among these genes, the ectopic expression of CaS gene increased salt tolerance in yeast, and CaS transcript was up-regulated under high salinity conditions. CaS-antisense transgenic plants showed reduced root elongation under 100 mM NaCl treatment compared to the wild type plant, which survived under 150 mM NaCl treatment, whereas CaS-antisense transgenic plant leaves turned yellow under 150 mM NaCl treatment. These results indicate that the expression of CaS gene is important for stress tolerance in yeast and plants.

Phenotypic and Transcriptomic Analysis of Nicotiana benthamiana Expressing Cucumber mosaic virus 2b gene (오이모자이크바이러스 2b 유전자 발현 담배의 형태 및 전사체 분석)

  • Sohn, Seong-Han;Kim, Yoon-Hee;Ahn, Yul-Kyun;Kim, Do-Sun;Won, So-Yoon;Kim, Jung-Sun;Choi, Hong-Soo
    • Research in Plant Disease
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    • v.21 no.3
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    • pp.186-192
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    • 2015
  • Cucumber mosaic virus possesses 2b gene known as a suppressor of post-transcriptional gene silencing (PTGS). To investigate its function and effect in plant, transgenic Nicotiana benethamiana expressing 2b gene was developed and analyzed in phenotypic characteristics and differential gene expression (DEG) comparing with wild-type. Eight lines of transgenic plants ($T_0$) were obtained with difficulty and showed severe deformed phenotypes in leaves, flowers, petioles and etc. Moreover, transgenic plants were hardly able to set seeds, but small amounts of seeds were barely produced in some of transgene-hemizygous plants. DEG analysis showed that transgenic plant ectopically accumulated diverse RNA transcripts at higher levels than wild-type probably due to the disturbance in RNA metabolism, especially of RNA decay, caused by 2b-mediated inhibition of PTGS. These ectopic accumulations of RNAs disrupt protein and RNA homeostasis and then subsequently lead to abnormal phenotypes of transgenic plants.

Functional characterization of a CCCH type zinc-finger protein gene OsZF2 by ectopic overexpression of the gene in rice (과발현 형질전환벼에서 CCCH type zinc-finger protein 유전자 OsZF2 기능 분석)

  • Lee, Jung-Sook;Yoon, In-Sun;Yoon, Ung-Han;Lee, Gang-Seob;Byun, Myung-Ok;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.23-29
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    • 2009
  • We have previously isolated a CCCH type zinc-finger protein gene, OsZF2 (Oryza sativa Zinc Finger 2), from the cold-treated rice cDNA library. To investigate the potential role of OsZF2, transgenic rice lines over-expressing OsZF2 under the control of CaMV 35S promoter have been developed through Agrobacterium-mediated transformation. Elevated level of OsZF2 transcripts was confirmed by RNA gel blot analysis in transgenic rice. Under the 100 mM NaCl condition, the transgenic rice showed significantly enhanced growth rate in terms of shoot length and fresh weight, implicating that OsZF2 is likely to be involved in salt response of rice. In the field condition, however, the transgenic rice showed a dwarf phenotype and flowering time was delayed. Genome expression profiling analysis of transgenic plants using the 20K NSF rice oligonucleotide array revealed many up-regulated genes related to stress responses and signaling pathways such as chaperone protein dnaJ 72, salt stress-induced protein, PR protein, disease resistance proteins RPM1 and Cf2/Cf5 disease resistance protein, carbohydrate/ sugar transporter, OsWAK kinase, brassinosteroid LRR receptor kinase, and jasmonate O-methyltransferase. These data suggest that the CCCH type zinc-finger protein OsZF2 is a upstream transcriptional factor regulating growth and stress responsiveness of rice.

Homeobox Gene (OSH1) Expression in Embryonic Mutants of Rice (Oryza sativa L.)

  • Hong, Soon-Kwan;Lee, Sang-Lyung;Shin, Young-Boum;Yoon, Kyung-Min;Kim, Nam-Soo
    • Animal cells and systems
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    • v.2 no.1
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    • pp.81-86
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    • 1998
  • Recent identification and characterization of plant homeobox genes suggest that they play important roles in morphogenetic events. OSH1, one of the rice homeobox genes, is thought to be related to organ development since the changes of OSH1 gene expression cause morphological abnormalities of leaves by the ectopic expression and is expressed during early embryogenesis. In this experiment, the expression pattern of OSH1 was analyzedinmutants by in situ hybridization, and OSH1's potential as a molecular marker was explored. Region-specific expression of OSH1 during early embryogenesis shows that OSH1 could be used as a molecular marker for characterizing embryo mutants. Although several organless and shootless mutants showed normal expression of OSM1, some mutants exhibited abnormal expression patterns. In a minute organless cle1-1 embryo whose epidermis resembled morphologically the epithelium of scutellum, OSH1 expression was limited to a small basal region. This expression pattern suggests the gross deletion of the basal part. In a radicleless mutant, odm115, OSH1 expression was detected in a basal region instead of subcentral region of the ventral side. Together with other characteristics (short embryo and normal adventitious roots), odm115 was estimated to be derived from the deletion of basal region. Among five shootless mutants, three showed normal expression of OSH1. In the shl2 embryo, no expression of OSH1 was observed. In the shl1 embryo, however, OSH1 expression was extended to a dorsal side, indicating that SHL2 might be related to dorsoventral patterning. The above results of in situ hybrydization clearly indicate that OSH1 can be utilized as a marker for characterizing gene functions of embryo mutants.

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IGF1 potentiates BMP9-induced osteogenic differentiation in mesenchymal stem cells through the enhancement of BMP/Smad signaling

  • Chen, Liang;Zou, Xiang;Zhang, Ran-Xi;Pi, Chang-Jun;Wu, Nian;Yin, Liang-Jun;Deng, Zhong-Liang
    • BMB Reports
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    • v.49 no.2
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    • pp.122-127
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    • 2016
  • Engineered bone tissue is thought to be the ideal alternative for bone grafts in the treatment of related bone diseases. BMP9 has been demonstrated as one of the most osteogenic factors, and enhancement of BMP9-induced osteogenesis will greatly accelerate the development of bone tissue engineering. Here, we investigated the effect of insulin-like growth factor 1 (IGF1) on BMP9-induced osteogenic differentiation, and unveiled a possible molecular mechanism underling this process. We found that IGF1 and BMP9 are both detectable in mesenchymal stem cells (MSCs). Exogenous expression of IGF1 potentiates BMP9-induced alkaline phosphatase (ALP), matrix mineralization, and ectopic bone formation. Similarly, IGF1 enhances BMP9-induced endochondral ossification. Mechanistically, we found that IGF1 increases BMP9-induced activation of BMP/Smad signaling in MSCs. Our findings demonstrate that IGF1 can enhance BMP9-induced osteogenic differentiation in MSCs, and that this effect may be mediated by the enhancement of the BMP/Smad signaling transduction triggered by BMP9.

Gelsolin Induces Promonocytic Leukemia Differentiation Accompanied by Upregulation of p21CIP1

  • Shirkoohi, Reza;Fujita, Hisakazu;Darmanin, Stephanie;Takimoto, Masato
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4827-4834
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    • 2012
  • Tumor suppressor genes have received much attention for their roles in the development of human malignancies. Gelsolin has been found to be down-regulated in several types of human cancers, including leukemias. It is, however, expressed in macrophages, which are the final differentiation derivatives for the monocytic myeloid lineage, implicating this protein in the differentiation process of such cells. In order to investigate the role of gelsolin in leukaemic cell differentiation, stable clones over-expressing ectopic gelsolin, and a control clone were established from U937 leukaemia cells. Unlike the control cells, both gelsolin-overexpressing clones displayed retarded growth, improved monocytic morphology, increased NADPH and NSE activities, and enhanced surface expression of the ${\beta}$-integrin receptor, CD11b, when compared with the parental U937 cells. Interestingly, RT-PCR and western blot analysis also revealed that gelsolin enhanced p21CIP1 mRNA and protein expression in the overexpressing clones. Moreover, transient transfection with siRNA silencing P21CIP1, but not the control siRNA, resulted in a reduction in monocytic differentiation, accompanied by an increase in proliferation. In conclusion, our work demonstrates that gelsolin, by itself, is capable of inducing monocytic differentiation in U937 leukaemia cells, most probably through p21CIP1 activation.