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opennotspecifiedNov 2020View details →

Fig. 2 in Bioactive diterpenoid metabolism and cytotoxic activities of genetically transformed Euphorbia lathyris roots

Fig. 2. Base peak chromatograms of transformed E. lathyris roots compared to wild-type plant roots and aerial parts with putatively assigned metabolites that are structurally related to ingenol.

opennotspecifiedNov 2020View details →

Fig. 4 in Bioactive diterpenoid metabolism and cytotoxic activities of genetically transformed Euphorbia lathyris roots

Fig. 4. Bioinformatic analysis and chemical elicitation of early diterpenoid biosynthetic genes expressed in transformed E. lathyris roots. Comparison of the deduced amino acid sequences of (a) ElFPS and (b) ElGGPS highlighting two conserved aspartate-rich domains [DDxx(xx)D]. (c) Comparison of the deduced amino acid sequence of ElCS highlighting a conserved [DDxxD] motif that is essential to the cyclization functionalities of terpene synthases. (d) Time course of E. lathyris diterpenoid biosynthetic gene transcript levels in transformed root cultures treated with 100 μM methyl jasmonate. Asterisks indicate statistical significance in comparison to 0 h control assessed by one-way ANOVA (**,P <0.01; *,P <0.05).

opennotspecifiedNov 2020View details →

Fig. 3 in Bioactive diterpenoid metabolism and cytotoxic activities of genetically transformed Euphorbia lathyris roots

Fig. 3. The MS/MS (fragmentation) data of m/z 477.2732 aided in the putative assignment of compound 8 as 15-O-acetyl-3-O-iso-butyryljolkinol-5β,6β-oxide. The fragmentation structures and m/z values correspond to each other (i.e. structures and peaks A-D).

Fig. 7 in Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity

Fig. 7. Molecular networking results from GNPS visualized with Cytoscape. Inset: cluster of UA and derivatives with close fragmentation pathway (m/z 357.12: compound K, m/z 389.104: compound L) and self-loop of compound H (at m/z 386.139).

Fig. 4 in Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity

Fig. 4. HPLC chromatograms of S. cyaneofuscatus cultures with or without UA: A) At the beginning of the stationary phase of bacterial growth; B) After 7 days of stationary phase. Circled in blue: compounds inhibited in the presence of UA, circled in orange: compounds more concentrated in the presence of UA, circled in red: UA. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 1 in Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity

Fig. 1. Monitoring of the bacterial growth over 15 days (D0 to D15) by measuring optical density (log OD (optical density), gray curve) and cell viability (%) using MTT assay (blue curve) compared to untreated culture (orange curve). A) Nocardia sp., B) S. cyaneofuscatus, C) M. ruber. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 3 in Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity

Fig. 3. HPLC chromatograms of Nocardia sp. culture with or without UA. A) At the beginning of the stationary phase of the bacterial growth; B) After 7 days of stationary phase. Compounds circled in red appear only in the culture with UA. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 2 in Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity

Fig. 2. HPLC chromatograms of M. ruber cultures with or without UA. A) At the beginning of the stationary phase of bacterial growth; B) After 7 days of stationary phase. Compounds circled in red appear only in the culture with UA. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 5 in Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity

Fig. 5. Comparison of HPLC chromatograms of B. weihenstephanensis extracts with UA at 0.01 mg/mL after 1 day of culture (blue), 9 days of culture (black) and without UA after 9 days of culture (red). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 8 in Lichen-associated bacteria transform antibacterial usnic acid to products of lower antibiotic activity

Fig. 8. Fragmentation patterns in negative mode of A) compound H and B) UA; common fragments are highlighted in orange. (For interpretation of the references to colour 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 2023View details →

Figure 5 in What causes transformation of the parasitic copepod? A new example of host switching in the family Anthessiidae (Cyclopoida) from Singaporean waters, with the proposal of a new genus

Figure 5. Merlionia zeeae, adult male, allotype (ZRC 2023.0306). A, habitus, dorsal. B, cephalosome, dorsal. C, rostral area, ventral. D, fifth pedigerous and genital somites, ventral. E, left antennule, anterior. F, left maxilliped, posterior. Scale bars: A, 400 μm; B, 200 μm; C, 50 μm; D–F, 100 μm.

opennotspecifiedJun 2023View details →

Figure 6 in What causes transformation of the parasitic copepod? A new example of host switching in the family Anthessiidae (Cyclopoida) from Singaporean waters, with the proposal of a new genus

Figure 6. Fresh coloration of a specimen of Ichthyscopus lebeck (Bloch & Schneider, 1801) infected by the type series of Merlionia zeeae. Scale bar: 30 mm.