Home About us Contact | |||
Transgenic Arabidopsis (transgenic + arabidopsi)
Terms modified by Transgenic Arabidopsis Selected AbstractsThe function of SULTR2;1 sulfate transporter during seed development in Arabidopsis thalianaPHYSIOLOGIA PLANTARUM, Issue 1 2005Motoko Awazuhara SULTR2;1 is a low-affinity sulfate transporter expressed in the vascular tissues of roots and leaves for interorgan transport of sulfate in Arabidopsis thaliana. Transgenic Arabidopsis carrying a fusion gene construct of SULTR2;1 5,-promoter region and ,-glucuronidase coding sequence (GUS) demonstrated that within the reproductive tissues, SULTR2;1 is specifically expressed in the bases and veins of siliques and in the funiculus, which connects the seeds and the silique. The antisense suppression of SULTR2;1 mRNA caused decrease of sulfate contents in seeds and of thiol contents both in seeds and leaves, as compared with the wildtype (WT). The effect of antisense suppression of SULTR2;1 on seed sulfur status was determined by introducing a sulfur-indicator construct, p35S::,SRx3:GUS, which drives the expression of GUS reporter under a chimeric cauliflower mosaic virus 35S promoter containing a triplicate repeat of sulfur-responsive promoter region of soybean ,-conglycinin , subunit (,SRx3). The mature seeds of F1 plants carrying both the SULTR2;1 antisense and p35S::,SRx3:GUS constructs exhibited significant accumulation of GUS activities on sulfur deficiency, as compared with those carrying only the p35S::,SRx3:GUS construct in the WT background. These results suggested that SULTR2;1 is involved in controlling translocation of sulfate into developing siliques and may modulate the sulfur status of seeds in A. thaliana. [source] Autohydrolysis of plant xylans by apoplastic expression of thermophilic bacterial endo-xylanasesPLANT BIOTECHNOLOGY JOURNAL, Issue 3 2010Bernhard Borkhardt Summary The genes encoding the two endo-xylanases XynA and XynB from the thermophilic bacterium Dictyoglomus thermophilum were codon optimized for expression in plants. Both xylanases were designed to be constitutively expressed under the control of the CaMV 35S promoter and targeted to the apoplast. Transient expression in tobacco and stable expression in transgenic Arabidopsis showed that both enzymes were expressed in an active form with temperature optima at 85 °C. Transgenic Arabidopsis accumulating heterologous endo-xylanases appeared phenotypically normal and were fully fertile. The highest xylanase activity in Arabidopsis was found in dry stems indicating that the enzymes were not degraded during stem senescence. High levels of enzyme activity were maintained in cell-free extracts from dry transgenic stems during incubation at 85 °C for 24 h. Analysis of cell wall polysaccharides after heat treatment of wildtype and transgenic extracts from dry stems showed a decrease in the molecular weight of xylans from transgenic stems. [source] Combining subproteome enrichment and Rubisco depletion enables identification of low abundance proteins differentially regulated during plant defensePROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 1 2009Ivy Widjaja Abstract Transgenic Arabidopsis conditionally expressing the bacterial avrRpm1 type III effector under the control of a dexamethasone-responsive promoter were used for proteomics studies. This model system permits study of an individual effector without interference from additional bacterial components. Coupling of different prefractionation approaches to high resolution 2-DE facilitated the discovery of low abundance proteins , enabling the identification of proteins that have escaped detection in similar experiments. A total of 34 differentially regulated protein spots were identified. Four of these (a remorin, a protein phosphatase 2C (PP2C), an RNA-binding protein, and a C2-domain-containing protein) are potentially early signaling components in the interaction between AvrRpm1 and the cognate disease resistance gene product, resistance to Pseudomonas syringae pv. maculicola 1 (RPM1). For the remorin and RNA-binding protein, involvement of PTM and post-transcriptional regulation are implicated, respectively. [source] AKIN,1 is Involved in the Regulation of Nitrogen Metabolism and Sugar Signaling in ArabidopsisJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 5 2009Xiao-Fang Li Abstract Sucrose non-fermenting-1-related protein kinase 1 (SnRK1) has been located at the heart of the control of metabolism and development in plants. The active SnRK1 form is usually a heterotrimeric complex. Subcellular localization and specific target of the SnRK1 kinase are regulated by specific beta subunits. In Arabidopsis, there are at least seven genes encoding beta subunits, of which the regulatory functions are not yet clear. Here, we tried to study the function of one beta subunit, AKIN,1. It showed that AKIN,1 expression was dramatically induced by ammonia nitrate but not potassium nitrate, and the investigation of AKIN,1 transgenic Arabidopsis and T-DNA insertion lines showed that AKIN,1 negatively regulated the activity of nitrate ruductase and was positively involved in sugar repression in early seedling development. Meanwhile AKIN,1 expression was reduced upon sugar treatment (including mannitol) and did not affect the activity of sucrose phosphate synthase. The results indicate that AKIN,1 is involved in the regulation of nitrogen metabolism and sugar signaling. [source] PPF1 May Suppress Plant Senescence via Activating TFL1 in Transgenic Arabidopsis PlantsJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 4 2008Da-Yong Wang Abstract Senescence, a sequence of biochemical and physiological events, constitutes the final stage of development in higher plants and is modulated by a variety of environmental factors and internal factors. PPF1 possesses an important biological function in plant development by controlling the Ca2+ storage capacity within chloroplasts. Here we show that the expression of PPF1 might play a pivotal role in negatively regulating plant senescence as revealed by the regulation of overexpression and suppression of PPF1 on plant development. Moreover, TFL1, a key regulator in the floral repression pathway, was screened out as one of the downstream targets for PPF1 in the senescence-signaling pathway. Investigation of the senescence-related phenotypes in PPF1(,) tfl1-1 and PPF1(+) tfl1-1 double mutants confirmed and further highlighted the relation of PPF1 with TFL1 in transgenic plants. The activation of TFL1 expression by PPF1 defines an important pathway possibly essential for the negative regulation of plant senescence in transgenic Arabidopsis. [source] Autohydrolysis of plant xylans by apoplastic expression of thermophilic bacterial endo-xylanasesPLANT BIOTECHNOLOGY JOURNAL, Issue 3 2010Bernhard Borkhardt Summary The genes encoding the two endo-xylanases XynA and XynB from the thermophilic bacterium Dictyoglomus thermophilum were codon optimized for expression in plants. Both xylanases were designed to be constitutively expressed under the control of the CaMV 35S promoter and targeted to the apoplast. Transient expression in tobacco and stable expression in transgenic Arabidopsis showed that both enzymes were expressed in an active form with temperature optima at 85 °C. Transgenic Arabidopsis accumulating heterologous endo-xylanases appeared phenotypically normal and were fully fertile. The highest xylanase activity in Arabidopsis was found in dry stems indicating that the enzymes were not degraded during stem senescence. High levels of enzyme activity were maintained in cell-free extracts from dry transgenic stems during incubation at 85 °C for 24 h. Analysis of cell wall polysaccharides after heat treatment of wildtype and transgenic extracts from dry stems showed a decrease in the molecular weight of xylans from transgenic stems. [source] The synthesis and accumulation of stearidonic acid in transgenic plants: a novel source of ,heart-healthy' omega-3 fatty acidsPLANT BIOTECHNOLOGY JOURNAL, Issue 7 2009Noemí Ruiz-López Summary Dietary omega-3 polyunsaturated fatty acids have a proven role in reducing the risk of cardiovascular disease and precursor disease states such as metabolic syndrome. Although most studies have focussed on the predominant omega-3 fatty acids found in fish oils (eicosapentaenoic acid and docosahexaenoic acid), recent evidence suggests similar health benefits from their common precursor, stearidonic acid. Stearidonic acid is a ,6-unsaturated C18 omega-3 fatty acid present in a few plant species (mainly the Boraginaceae and Primulaceae) reflecting the general absence of ,6-desaturation from higher plants. Using a ,6-desaturase from Primula vialii, we generated transgenic Arabidopsis and linseed lines accumulating stearidonic acid in their seed lipids. Significantly, the P. vialii,6-desaturase specifically only utilises ,-linolenic acid as a substrate, resulting in the accumulation of stearidonic acid but not omega-6 ,-linolenic acid. Detailed lipid analysis revealed the accumulation of stearidonic acid in neutral lipids such as triacylglycerol but an absence from the acyl-CoA pool. In the case of linseed, the achieved levels of stearidonic acid (13.4% of triacylglycerols) are very similar to those found in the sole natural commercial plant source (Echium spp.) or transgenic soybean oil. However, both those latter oils contain ,-linolenic acid, which is not normally present in fish oils and considered undesirable for heart-healthy applications. By contrast, the stearidonic acid-enriched linseed oil is essentially devoid of this fatty acid. Moreover, the overall omega-3/omega-6 ratio for this modified linseed oil is also significantly higher. Thus, this nutritionally enhanced linseed oil may have superior health-beneficial properties. [source] The DDF1 transcriptional activator upregulates expression of a gibberellin-deactivating gene, GA2ox7, under high-salinity stress in ArabidopsisTHE PLANT JOURNAL, Issue 4 2008Hiroshi Magome Summary High-salinity stress affects plant growth and development. We have previously reported that overexpression of the salinity-responsive DWARF AND DELAYED FLOWERING 1 (DDF1) gene, encoding an AP2 transcription factor of the DREB1/CBF subfamily, causes dwarfism mainly by levels of reducing bioactive gibberellin (GA) in transgenic Arabidopsis. Here, we found that the GA 2-oxidase 7 gene (GA2ox7), which encodes a C20 -GA deactivation enzyme, is strongly upregulated in DDF1 -overexpressing transgenic plants. A loss-of-function mutation of GA2ox7 (ga2ox7-2) suppressed the dwarf phenotype of DDF1 -overexpressing plants, indicating that their GA deficiency is due to overexpression of GA2ox7. Transient overexpression of DDF1 activated the promoter of GA2ox7 in Arabidopsis leaves. A gel shift assay showed that DDF1 binds DRE-like motifs (GCCGAC and ATCGAC) in the GA2ox7 promoter. In Arabidopsis under high-salinity stress, six GA2ox genes, including GA2ox7, were upregulated. Furthermore, the ga2ox7-2 mutant was less growth retarded than wild-type Col under high-salinity stress. These results demonstrate that, under salinity stress, Arabidopsis plants actively reduce endogenous GA levels via the induction of GA 2-oxidase, with the result that growth is repressed for stress adaptation. [source] Identification of a novel cis -acting element conferring sulfur deficiency response in Arabidopsis rootsTHE PLANT JOURNAL, Issue 3 2005Akiko Maruyama-Nakashita Summary SULTR1;1 high-affinity sulfate transporter is highly regulated in the epidermis and cortex of Arabidopsis roots responding to sulfur deficiency (,S). We identified a novel cis -acting element involved in the ,S-inducible expression of sulfur-responsive genes in Arabidopsis. The promoter region of SULTR1;1 was dissected for deletion and gain-of-function analysis using luciferase (LUC) reporter gene in transgenic Arabidopsis. The 16-bp sulfur-responsive element (SURE) from ,2777 to ,2762 of SULTR1;1 promoter was sufficient and necessary for the ,S-responsive expression, which was reversed when supplied with cysteine and glutathione (GSH). The SURE sequence contained an auxin response factor (ARF) binding sequence (GAGACA). However, SURE was not responsive to naphthalene acetic acid, indicating its specific function in the sulfur response. The base substitution analysis indicated the significance of a 5-bp sequence (GAGAC) within the conserved ARF binding site as a core element for the ,S response. Microarray analysis of early ,S response in Arabidopsis roots indicated the presence of SURE core sequences in the promoter regions of ,S-inducible genes on a full genome GeneChip array. It is suggested that SURE core sequences may commonly regulate the expression of a gene set required for adaptation to the ,S environment. [source] Biochemical and functional analysis of CTR1, a protein kinase that negatively regulates ethylene signaling in ArabidopsisTHE PLANT JOURNAL, Issue 2 2003Yafan Huang Summary CTR1 encodes a negative regulator of the ethylene response pathway in Arabidopsis thaliana. The C-terminal domain of CTR1 is similar to the Raf family of protein kinases, but its first two-thirds encodes a novel protein domain. We used a variety of approaches to investigate the function of these two CTR1 domains. Recombinant CTR1 protein was purified from a baculoviral expression system, and shown to possess intrinsic Ser/Thr protein kinase activity with enzymatic properties similar to Raf-1. Deletion of the N-terminal domain did not elevate the kinase activity of CTR1, indicating that, at least in vitro, this domain does not autoinhibit kinase function. Molecular analysis of loss-of-function ctr1 alleles indicated that several mutations disrupt the kinase catalytic domain, and in vitro studies confirmed that at least one of these eliminates kinase activity, which indicates that kinase activity is required for CTR1 function. One missense mutation, ctr1,8, was found to result from an amino acid substitution within a new conserved motif within the N-terminal domain. Ctr1,8 has no detectable effect on the kinase activity of CTR1 in vitro, but rather disrupts the interaction with the ethylene receptor ETR1. This mutation also disrupts the dominant negative effect that results from overexpression of the CTR1 amino-terminal domain in transgenic Arabidopsis. These results suggest that CTR1 interacts with ETR1 in vivo, and that this association is required to turn off the ethylene-signaling pathway. [source] |