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34 results for “Lotus japonicus”
Identification of novel genes involved in phosphate accumulation in Lotus japonicus through Genome Wide Association mapping of root system architecture and anion content
<p>130 Lotus japonicus accessions were used. The names and accession numbers are<br> listed in S6 Table. Seeds were scarified with sandpaper and then sterilized 14 minutes in 0.05%<br> sodium hypochlorite. Subsequently, seeds were rinsed and washed 5 times in sterile distilled<br> water. For the germination, seeds were positioned in imbibed filter paper, in sterile Petri dishes,<br> and wrapped in aluminium foil. After 3 days at 21°C, young seedling were transferred to square<br> plates (12 x 12 cm) containing growth medium. Both media used in this<br> study were based on Long-Ashton solution (with two levels of phosphate concentration -20 or<br> 750 μM, LP or HP, respectively) with 0.8% MES buffer (Duchefa Biochemie,<br> Haarlem, The Netherlands), 0.8% agarose (to minimize phosphate contamination), and adjusted<br> to pH 5.7 with 1M KOH. After adding the medium, plates were dried, closed, overnight in a<br> sterile laminar flow hood. Two accessions, with four replicates per each accession, were placed<br> on each plate. Each plate was replicated, with mirrored position of each accession to minimize<br> any positional growth effects. Plates were placed vertically, and plants grown under long-day<br> conditions (21°C, 16 h light/8 h dark cycle) with white light bulbs emitting 50 μmol/m 2 /s and<br> roots were exposed to light. Every day at the same time, the racks were transported to the image<br> acquisition room where images of each plate were acquired with eight Epson V600 CCD flatbed<br> color image scanners (Seiko Epson) and then immediately returned to the growth chamber.</p>
Nitrogen fertilization nullifies host sanctions against non-fixing rhizobia and drives divestment from symbiosis in Lotus japonicus
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Fig. 8 in Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues
Fig. 8. Effect of overexpression of different LjSK1 variants in L. japonicus hairy roots. Physiological parameters of L. japonicus wild type hairy roots transformed with LjSK1 variants: LjSK1 90–467 lacking the 89 N-terminal residues, LjSK1_K167A, LjSK1_Y298A, and native LjSK1 at 21 days post inoculation with rhizobium. Control roots were transformed with the empty T-DNA vector. Values shown are the average ± SEM of n = 6–16 individual hairy roots per construct. Letters above each graph indicate statistically significant differences (p <0.05) between samples using One-way ANOVA followed by Fischer's LSD post-hoc comparison.
Fig. 7 in Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues
Fig. 7. Appearance of whole plants and nodules of L. japonicus hairy root lines transformed with the different LjSK1 variants. A, F) Control (empty vector); B, G) native LjSK1; C, H) LjSK1 K167A; D, I) LjSK1 Y298A; E, J) LjSK190-467. A-E) Representative whole plants with regenerated hairy roots. F-J) Images of nodules on regenerated hairy roots taken in bright-field (top panels) and with an RFP filter to visualize rhizobium colonisation (bottom panels). Scale bars in A-E are 5 cm and in F-G 2 mm.
Fig. 6. Western blot analysis with a p in Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues
Fig. 6. Western blot analysis with a p-GSK-3α/β (6D3) (Santa Cruz Biotechnology, Inc.) monoclonal antibody raised against a phosphopeptide corresponding to amino acids residues surrounding Tyr279 of human GSK3α and Tyr216 of human GSK3β. Lane 1: recombinant LjSK1 and lane 2: LjSK1 Y298A variant. Both proteins have a GST-tag.
Fig. 4 in Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues
Fig. 4. pH (A) and temperature (B) curves for LjSK1 together with standard error bars. Points represent the average value of three measurements. RLU refers to Relative Light Units and ΔRLU is for the difference between a control sample (without enzyme) and the corresponding sample with enzyme. Measurements were performed as described in section 5.3.
Fig. 2. A in Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues
Fig. 2. A homology model of the LjSK1 – ATP complex based on the structure of human GSK3β (PDB:1J1B) (Aoki et al., 2004) after refinement with molecular dynamics simulations and docking of the ATP molecule at the ATP binding site. N- and C-terminal lobes are indicated by different colors and the G-loop and the activation loop are indicated in purple and green, respectively. Mutated residues are shown as stick models. Figure created using Pymol (DeLano, 2002). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 1 in Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues
Fig. 1. Alignment of the primary structure of LjSK1, human GSK3β and AtSK3-2. Identical residues are shown in bold. Residues mutated to alanine are boxed. Residues constituting G-loop and the activation loop are shown in grey boxes and the respective structural feature is indicated. Black arrows indicate residues predicted to be in close proximity to the phosphates of ATP. Gaps, represented by dashes, were introduced into the sequences to optimize the alignment.
Fig. 7 in Chemical identification of 18-hydroxycarlactonoic acid as an LjMAX1 product and in planta conversion of its methyl ester to canonical and noncanonical strigolactones in Lotus japonicus
Fig. 7. Conversion of [13C]-18-OH-MeCLA to [13C]-5DS and [13C]-lotuslactone (LL) in the feeding experiment using L. japonicus roots. A) Conversion of [10–13C]-18-OH-MeCLA to [6′-13C]-5DS. LC-MS/MS analysis of [6′-13C]-5DS in root exudates after feeding [10–13C]-18-OHMeCLA. MRM chromatograms (left) and full-scan spectra of fragment ions (right). The MRM chromatograms of authentic 5DS (red: 331.15/217.00, blue: 331.15/97.00, green: 331.15/234.00, m/z in positive mode) and [6′-13C]-5DS (red: 332.15/218.00, blue: 332.15/97.00, green: 332.15/ 235.00, m/z in positive mode) are shown. B) Conversion of [10–13C]-18- HO-MeCLA to [6′-13C]-LL. LC-MS/MS analysis of [6′-13C]-LL in root exudates after feeding [10–13C]-18-OH-MeCLA. MRM chromatograms (left) and full-scan spectra of fragment ions (right). The MRM chromatograms of authentic LL (red: 373.00/276.00, blue: 373.00/244.00, green: 373.00/ 216.00, m/z in positive mode) and [6′-13C]-LL (red: 374.00/277.00, blue: 374.00/245.00, green: 374.00/216.00, m/z in positive mode) are shown. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 8 in Chemical identification of 18-hydroxycarlactonoic acid as an LjMAX1 product and in planta conversion of its methyl ester to canonical and noncanonical strigolactones in Lotus japonicus
Fig. 8. The proposed biosynthetic pathway of 5DS and LL in L. japonicus. LjMAX1 catalyzes the oxidation of CL to 18-OH-CLA via CLA. (11R)-CL, CLA and 18- hydroxylated carlactonoates are precursors for 5DS and LL in L. japonicus. Solid arrows indicate confirmed pathways reported in previous studies and this study and dashed arrows indicate putative pathways. Blue letters indicate the position number. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 6 in Chemical identification of 18-hydroxycarlactonoic acid as an LjMAX1 product and in planta conversion of its methyl ester to canonical and noncanonical strigolactones in Lotus japonicus
Fig. 6. Conversion of [13C]-CLA to [13C]-5DS and [13C]-18-OH-CLA in the feeding experiment using L. japonicus roots. A) Conversion of [1-13CH]- 3 CLA to [8-13CH]-5DS. LC-MS/MS analysis of [8-13CH]-5DS in root exu3 3 dates after feeding [1-13CH]-CLA. MRM chromatograms (left) and full3 scan spectra of fragment ions (right). The MRM chromatograms of authentic 5DS (red: 331.15/217.00, blue: 331.15/97.00, green: 331.15/ 234.00, m/z in positive mode) and [8-13CH]-5DS (red: 332.15/218.00, 3 blue: 332.15/97.00, green: 332.15/235.00, m/z in positive mode) are shown. B) Conversion of [1-13CH]-CLA to [1-13CH]-18-OH-CLA. LC-MS/ 3 3 MS analysis of [1-13CH]-18-OH-CLA in root exudates after feeding 3 [1-13CH]-CLA. MRM chromatograms (left) and full-scan spectra of frag3 ment ions (right). Authentic 18-OH-CLA was prepared by feeding CL to recombinant Os900. MRM chromatograms of authentic 18-OH-CLA (red: 347.00/303.00, blue: 347.00/113.00, m/z in negative mode) and [1-13CH]-18-OH-CLA (red: 348.00/304.00, blue: 348.00/113.00, m/z in 3 negative mode) are shown. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 3 in Chemical identification of 18-hydroxycarlactonoic acid as an LjMAX1 product and in planta conversion of its methyl ester to canonical and noncanonical strigolactones in Lotus japonicus
Fig. 3. Synthetic scheme of 13C-labeled and unlabeled 18-OH-MeCLA. Asterisks indicate the position of 13C.
Fig. 2 in Chemical identification of 18-hydroxycarlactonoic acid as an LjMAX1 product and in planta conversion of its methyl ester to canonical and noncanonical strigolactones in Lotus japonicus
Fig. 2. Detection of 18-hydroxycarlactonoic acid (18-OH-CLA), 4DO and 5DS in CLA feeding experiment by recombinant LjMAX1 and Os900. CLA was incubated with recombinant yeast microsomes. Yeast microsomes having an empty vector and the expression vector pYeDP60-Os900 were used as a negative and a positive control, respectively. The extracts of the microsomes and authentic standard were analyzed by LC-MS/MS. The peak at 7.0 min was presumed to be 18-OH-CLA which may be converted artificially to 4DO and 5DS in the ion source of mass spectrometer, and detected in the MRM transitions for 4DO and 5DS (red: 331.15/216.00, blue: 331.15/97.00, green: 331.15/234.00, m/z in positive mode). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 1 in Chemical identification of 18-hydroxycarlactonoic acid as an LjMAX1 product and in planta conversion of its methyl ester to canonical and noncanonical strigolactones in Lotus japonicus
Fig. 1. Detection of carlactonoic acid (CLA), 4-deoxyorobanchol (4DO) and 5- deoxystrigol (5DS) in carlactone (CL) feeding experiment by recombinant Lotus japonicus MAX1 (LjMAX1) and Os900. CL was incubated with yeast microsomes. Yeast microsomes having an empty vector and the expression vector pYeDP60-Os900 were used as a negative and a positive control, respectively. The extracts of the microsomes and authentic standard were analyzed by LCMS/MS. MRM chromatograms of CLA (red: 331.10/69.00, blue: 331.10/ 113.00, m/z in negative mode), 4DO and 5DS (red: 331.15/216.00, blue: 331.15/97.00, green: 331.15/234.00, m/z in positive mode) are shown. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig. 5 in Mutagenesis of a Lotus japonicus GSK3β/Shaggy-like kinase reveals functionally conserved regulatory residues
Fig. 5. The enzymatic activity of native LjSK1. The inset shows the Lineweaver- Burk plot.
A time-course transcriptomic study for investigating the genome-wide effects of nitrate on gene expression in mature nodules of Lotus japonicus.
GEO Series GSE197362. Lotus japonicus. 36 samples. Type: Expression profiling by high throughput sequencing.
Constitutive over-expression of LjMyb14 in Lotus japonicus.
GEO Series GSE31739. Lotus japonicus. 6 samples. Type: Expression profiling by array.
Ectopic expression of NAC094 induces premature nodule senescence in Lotus japonicus.
GEO Series GSE197361. Lotus japonicus. 6 samples. Type: Expression profiling by high throughput sequencing.
Transcriptome analysis of Arabidopsis thaliana and Lotus japonicus exposed to arbuscular mycorrhizae
GEO Series GSE225213. Arabidopsis thaliana; Lotus japonicus. 44 samples. Type: Expression profiling by high throughput sequencing.
Tissue-specific transcriptome analysis in nodules of Lotus japonicus
GEO Series GSE34753. Lotus japonicus. 9 samples. Type: Expression profiling by array.
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