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opennotspecifiedOct 2020View details →

Fig. 3 in Natural variation in specialised metabolites production in the leafy vegetable spider plant (Gynandropsis gynandra L. (Briq.)) in Africa and Asia

Fig. 3. Heatmap of 107 significant semi-polar metabolites with high PCA loadings (>|0.7|) in 48 accessions of Gynandropsis gynandra from Asia (red), East/Southern Africa (black) and West Africa (blue). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

opennotspecifiedOct 2020View details →

Fig. 2 in Natural variation in specialised metabolites production in the leafy vegetable spider plant (Gynandropsis gynandra L. (Briq.)) in Africa and Asia

Fig. 2. Principal component analysis score plot of relative levels of 936 semi-polar metabolites detected in the leaves of 48 accessions of Gynandropsis gynandra from Asia (red), East/Southern Africa (black) and West Africa (blue). The first two dimensions explaining 39.6% of the total variation are shown. 95% confidence ellipses are displayed for the three regions. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 6 in Variation in production of cyanogenic glucosides during early plant development: A comparison of wild and domesticated sorghum

Fig. 6. Proportion of nitrogen allocated to dhurrin and nitrate (NO3) in dried, finely ground tissues of S. bicolor, S. brachypodum and S. macrospermum plants at 35 d post-germination. A) Dhurrin allocation; B) Nitrate allocation; C) C:N ratio. Graphs show mean ± 1 standard error (n = 3). Columns with different letters within each tissue are significantly different (p <0.05).

Fig. 3 in Variation in production of cyanogenic glucosides during early plant development: A comparison of wild and domesticated sorghum

Fig. 3. Leaf characteristics of S. bicolor, S. brachypodum and S. macrospermum plants at six harvest points during the first 35 d post-germination. A) Total leaf number; B) Total leaf area (TLA); C) Specific leaf area (SLA); D) Leaf area ratio (LAR). Graphs show mean ± 1 standard error (n = 5), with statistically significant differences indicated at each time point: *p <0.05.

Fig. 5 in Variation in production of cyanogenic glucosides during early plant development: A comparison of wild and domesticated sorghum

Fig. 5. Tissue-specific hydrogen cyanide potential (HCNp, mg HCN per g dw 1) and morphology of individual A) S. bicolor, B) S. brachypodum and C) S. macrospermum plants at six time points during seedling development. The HCNp of a section of the sheath, roots, and each individual leaf was measured at 3, 7, 14, 21, 28 and 35 days (D) post-germination. Colour scale indicates HCNp, used as a proxy for dhurrin concentration (green = low; red = high). (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4 in Variation in production of cyanogenic glucosides during early plant development: A comparison of wild and domesticated sorghum

Fig. 4. Hydrogen cyanide potential (HCNp, mg HCN per g dw 1) and concentration of nitrate (NO) in dried, finely ground tissues of S. bicolor, S. brachypodum and 3 S. macrospermum plants at six harvest points during the first 35 d post-germination (at 35 dpg only for NO3). A) Leaf HCNp; B) Sheath HCNp; C) Root HCNp; D) Total NO3. Graphs show mean ± 1 standard error (n = 5), with statistically significant differences indicated at each time point: *p <0.05. Columns with different letters within each tissue are significantly different (p <0.05). Data for leaf HCNp at 3 days post-germination not shown as a true leaf had not emerged at this stage.

Fig. 1 in Variation in production of cyanogenic glucosides during early plant development: A comparison of wild and domesticated sorghum

Fig. 1. Known geographic distribution of the two wild Sorghum species S. brachypodum and S. macrospermum and the site of collection of the accessions examined in the current study. Seeds were obtained from the Australian Grains Genebank (AGG), Horsham, Victoria. Occurrence records of S. brachypodum and S. macrospermum were obtained from the Atlas of Living Australia (ALA), htt p://www.ala.org.au. Each blue circle represents an occurrence record of S. brachypodum and each orange circle represents S. macrospermum (circled). Collection localities of individual accessions examined here are marked by darker coloured circles. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4. A in Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds

Fig. 4. A. Growth of wild type root cultures on different Trp derivatives as compared to growth on MS medium only. B. Correlation of the relative growth and the production of different Cl-Trp compounds.

Fig. 3. A in Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds

Fig. 3. A. All transgenic root lines were analyzed by the following experiments and the data are presented for selected lines transformed with the pMDC32+2xCaMV35S:pyrH:nosT construct (pyrH = 5-Cl-Trp-forming). Transformation of roots with A. rhizogenes was verified using gDNA and cDNA for a successful insertion and expression, respectively. A. Upper panel: Amplified rolB (423 bp) and rolC (626 bp) for three different lines (lanes 1–3) using gDNA. The virG gene (350 bp) was only detectable in the positive control (Ri-plasmid of A. rhizogenes) (lane +) "-" denotes a negative PCR control. Lower panel: Integration of full length hal gene (ca. 1.5 kb) using gDNA. Expression of full length hal gene (ca. 1.5 kb) using cDNA. "+": positive control (plasmid containing pyrH or the other hal genes), "-": negative PCR control, g: gDNA wild type, c: cDNA wild type, 1–3: three independent transgenic root culture lines with the pMDC32+2xCaMV35S:pyrH:nosT construct, "1-"-"3-": RT negative controls (containing no DNA). B. Western blot with the purified His-tagged proteins: 1: PyrH, 2: ThaI, 3: PrnA, a: PyrH synthesized in E. coli, b: positive control ThaI synthesized in E. coli, c: positive control PrnA synthesized in E. coli. Wild type protein as control did not show any signal (data not shown). C. Enzyme assay with the purified halogenase PyrH. The positive control is PyrH protein synthesized in bacteria. The negative control is purified protein from wild type root cultures. Since only for PyrH enzyme activity could be detected, the data for the other halogenases are shown in the supplement (Fig. S1). D. Production of chlorinated tryptophan (Cl-Trp) and indole-3-acetonitrile (ClIAN) in transgenic root lines. For each halogenase construct five independent lines were tested. Results for 5 lines per halogenase type with and without Histag are indicated by the numbers of lines with the respective metabolites. The detailed results for all individual lines are shown in the supplement (Fig. S2).

Fig. 6. A in Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds

Fig. 6. A. Confirmation of i) that the regenerated plants contain still the rol genes and ii) the integration of the hal gene into the genome and its transcription into cDNA (1: regenerated plants from wild type root cultures; 2: BrRP-pyrHHIS.6; 3: A. rhizogenes plasmid; 4: negative PCR control; 5: positive control - hal amplification from plasmid; a: cDNA, b: cDNA "no template control; genomic DNA. B. Western blot of His-tagged halogenase (PyrH, Thal, PrnA: purified enzymes from overexpressing E. coli strain as positive controls; BrRP-HR = WT, regenerated plants from wild type root cultures; BrRP-S: regenerated plants from Chinese cabbage seedlings; BrRP-pyrH, -thal, -prnA: regenerated plants from transgenic roots.). Always two different dilutions were applied. The gel strips were from the same gel, but due to large parts with samples without an immunosignal, the respective areas were cut out and are presented here. C. Relative amounts of chlorinated metabolites in the regenerated plants.

Fig. 2 in Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds

Fig. 2. Expected indole metabolites and their interconversion (in black) that could be derived from tryptophan via the indole glucosinolate/indole phytoalexin pathway. It is indicated (in grey) that there are alternative pathways to IAA. The possible induction (dashed arrows) of chlorinated metabolites by abiotic and biotic stress factors, the latter also via the signaling molecules salicylic acid and jasmonic acid, is shown.

Fig. 1 in Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds

Fig. 1. Experimental scheme showing the different types of plant materials generated. A. Mature wild type plants/seedlings; B. Wild type and transgenic root cultures; C. Regenerated sterile plants from wild type seedlings; D. Regenerated sterile plants from wild type and transgenic root cultures; E. Adult plants in soil from wild type cultures; F. Adult plants in soil from transgenic root cultures.

Fig. 5 in Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds

Fig. 5. Left: Average number of regenerated shoots for 25 Brassica rapa "hairy root" lines (denoted therefore as BrHR ….) on 3 media compositions (n = 24). Medium A: GB5 medium containing 8 g l−1 phytoagar, 20 g l−1 sucrose and 10 mg l−1 6-BAP. Medium B: MS medium containing 8 g l−1 phytoagar, 30 g l−1 sucrose, 4 mg l−1 6-BAP, 4 mg l−1 AgNO3 and 3 mg l−1 NAA. Medium C: MS medium containing 8 g l−1 phytoagar, 30 g l−1 sucrose, 4 mg l−1 6-BAP, 4 mg l−1 AgNO3 and 0.5 mg l−1 NAA. Right: Shoot regeneration from B. rapa root cultures. Pieces from these root cultures were cut into pieces of approximately 1 cm2 and placed on semisolid agar (A). Regeneration of shoots was visible after 4 weeks of cultivation (B). Regenerated shoots were separated and transferred to fresh media (C). Shoot growth was often accompanied by growth of transformed/transgenic roots (D). Shoots of adequate biomass quality were subcultivated (E). Some B. rapa lines displayed a shortened life cycle after regeneration and began flowering (F).

Fig. 7 in Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds

Fig. 7. Comparison of phenotypic traits for three groups of Brassica rapa grown in the greenhouse. WT – shoots from wild type plants grown from seeds (photo A); REG – regenerated shoots originated from transformed root cultures (photo B); HLR – regenerated shoots originated from transgenic roots transfected with bacterial hal genes (photo C). Significant differences between treatments are labeled as follows: 0 '***' 0.001 '**' 0.01 '*' 0.05 (with n = minimum of 20 individually potted plants). Leaves of in vitro shoots originated from seeds (left) or regenerated from root cultures (right) are shown.

opennotspecifiedJun 2020View details →

Fig. 4 in Pilot study on the uptake and modification of harmaline in acceptor plants: An innovative approach to visualize the interspecific transfer of natural products

Fig. 4. Confocal laser-scanning microscopy of a barley leaf incubated with harmaline. Five days after application of the alkaloid, leaves were examined by confocal laser-scanning microscopy (λex = 364 nm). The fluorescence spectra had been recorded for different areas or cell types, respectively, which are displayed by coloured circles; red: xylem; green and blue mesophyll. The analogous spectra are presented in the corresponding colour. According to the wavelength of maximal emission, the fluorescence could be attributed to lignin (xylem), harmaline (cells in the proximity of the vascular bundles) and harmine (cells more distant from them). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

opennotspecifiedJun 2020View details →

Fig. 1 in Pilot study on the uptake and modification of harmaline in acceptor plants: An innovative approach to visualize the interspecific transfer of natural products

Fig. 1. Coloration of pea seedlings after application of red beet extract to the culture medium. Red beet extract was applied to healthy, non-injured etiolated pea seedlings grown in a Petri dish in a dark room at the lab bench as described in the Experimental section. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)