Find research datasets worth reusing

Fig. 5. NtCOMT1 in NtCOMT1 responsible for phytomelatonin biosynthesis confers drought tolerance in Nicotiana tabacum

Fig. 5. NtCOMT1 gene expression profiles in different tissues (A) and under abiotic stresses (B). Growth of NtCOMT1 overexpressing lines, WT, and OE-Empty tobacco after drought treatment for 9 days (C). Expression levels in different tissues are shown: root, stem, leaf, flower, and fruit. Expression levels under cold stress (3h, 6h, and 24h), heat stress (3h, 6h, and 9h), drought stress (1day, 5day, and 9day), and cadmium stress treatment (1day, 4day, and 7day). Data represent mean ± SE of three independent biological replicates. Asterisks indicate statistically significant differences determined using Student' s t-test (**, p <0.01; ***, p <0.001).

Fig. 4 in NtCOMT1 responsible for phytomelatonin biosynthesis confers drought tolerance in Nicotiana tabacum

Fig. 4. Subcellular localization of the NtCOMT1 proteins in N. benthamiana leaves. (A) 35S-GFP alone was used as the control. (B) NtCOMT1 proteins fused with GFP (35S: NtCOMT1-GFP). (C) Fluorescence of DAPI. (D) merge of 35S: NtCOMT1-GFP and DAPI images.

Fig. 3 in NtCOMT1 responsible for phytomelatonin biosynthesis confers drought tolerance in Nicotiana tabacum

Fig. 3. Characterization of NtCOMT1 enzymatic activity. (A) Schematic diagram of melatonin biosynthesis mediated by COMT, converting N-acetylserotonin into melatonin. (B) Purification of His × 6-tagged NtCOMT1, M, molecular mass standards; lane 1, total proteins in 20-μL aliquots of bacterial culture with IPTG; lane 2, 20-μL aliquots of supernatant derived from bacterial cell lysate; lane 3, 20-μL aliquots of precipitated cell lysate; lane 4, purified NtCOMT1 protein. (C) The enzymatic activity of NtCOMT1 under different pH and temperature. (D) Kinetic analysis of purified recombinant NtCOMT1. (E) HPLC analysis of in vitro enzyme activity using N-acetylserotonin as the substrate. The retention time of the new peak is the same as the retention time of the melatonin standard (top: melatonin standard, middle: N-acetylserotonin standard, NtCOMT1: the corresponding protein).

Fig. 2 in NtCOMT1 responsible for phytomelatonin biosynthesis confers drought tolerance in Nicotiana tabacum

Fig. 2. Multiple sequence alignment of tobacco COMT as well as well-defined plants COMT proteins. Residues involved in SAM binding (pink), substrate binding (blue), substrate binding in transform the dyad-related polypeptide (green), and catalysis (red) are highlighted. (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 NtCOMT1 responsible for phytomelatonin biosynthesis confers drought tolerance in Nicotiana tabacum

Fig. 1. Phylogenetic relationships (A) and motif compositions (B) of tobacco COMT as well as well-defined plant COMT proteins. The phylogenetic tree was constructed with a bootstrap of 1000 by the maximum-likelihood method. Motif compositions were illustrated by TBtools. The proteins in phylogenetic tree include wellcharacterized OsCOMT (Oryza satica: LOC_Os09g17560), AtCOMT (Arabidopsis thaliana: AT5G54160), SlCOMT (Solanum lycopersicum: XP_004235028), CrCOMT (Carex rigescens: QDF21518), TaCOMT (Triticum aestivum: Traes_1AL_D9035D5E0), and GhCOMT (Gossypium hirsutum: Gh_D12G2714).

Fig. 6 in A highly efficient, thermo stable and broad pH adaptable copper-zinc super oxide dismutase (AmSOD1) mediates hydrogen peroxide tolerance in Avicennia marina

Fig. 6. Longevity of AmSOD1 activity. Purified His-AmSOD1 was tested for its longevity both at room temperature and 4 ◦ C for different time period (20, 40, 60, 80, 100, 120 1st 140 days). The ratio enzyme activity was determined based on the activity of enzyme before incubation. Each data point represents the mean ± SD obtained from three independent reactions.

Fig. 7 in A highly efficient, thermo stable and broad pH adaptable copper-zinc super oxide dismutase (AmSOD1) mediates hydrogen peroxide tolerance in Avicennia marina

Fig. 7. Growth curves of the strains control and R-AmSOD1 in the medium without treatment and the medium containing containing (A) salt and (B) hydrogen peroxide. Each data point represents the mean ± SD obtained from three independent reactions.

Fig. 2 in A highly efficient, thermo stable and broad pH adaptable copper-zinc super oxide dismutase (AmSOD1) mediates hydrogen peroxide tolerance in Avicennia marina

Fig. 2. The effects of salt and hydrogen peroxide stress on the appearance pattern and activity of AmSOD1 in the leaves of A. marina (A) Appearance pattern of AmSOD1 and (B) AmSOD assay in the leaves responding to hydrogen peroxide and salt stress as well as control at various time points (0–48 h). Each histogram represents the mean obtained from three independent reactions (± SD). Different letters above the bars indicate statistically significant differences by LSD test.

Fig. 1 in A highly efficient, thermo stable and broad pH adaptable copper-zinc super oxide dismutase (AmSOD1) mediates hydrogen peroxide tolerance in Avicennia marina

Fig. 1. Phylogenetic tree of SOD sequences from different plants. For accession numbers and organisms, see ''Materials and Methods.''.

Fig. 4 in A highly efficient, thermo stable and broad pH adaptable copper-zinc super oxide dismutase (AmSOD1) mediates hydrogen peroxide tolerance in Avicennia marina

Fig. 4. Molecular mass of purified AmSOD1 using gel filtration. (A) Native molecular mass of recombinant AmSOD1 was measured with respect to standard protein marker as resolved in Biosil-250 analytical gel filtration column. Standard proteins used were: (a) 66.5 kDa-BSA fraction (b) 29 kDa-Carbonic anhydrase and (c) 16.7 kDa human myoglubin.

Fig. 3 in A highly efficient, thermo stable and broad pH adaptable copper-zinc super oxide dismutase (AmSOD1) mediates hydrogen peroxide tolerance in Avicennia marina

Fig. 3. SDS-PAGE analysis and the solubility of expressed His-AmSOD1 in the strain R-AmSOD1 harboring pET28a-AmSOD1 (A) Total soluble protein extracted from control strain and the strain R-AmSOD1 before and 1–4 h after induction with IPTG. (B) The study of solubility of His-AmSOD1 by comparison of His-AmSOD1 band intensities in the total protein (T), the soluble fraction extracted from R-AmSOD1 (S) and the unsoluble fraction (U) extracted from RAmSOD1 (The recombinant proteins were shown with arrows) (C) SDS-PAGE analysis of AmSOD1 after purification with affinity chromatography.

Fig. 5 in A highly efficient, thermo stable and broad pH adaptable copper-zinc super oxide dismutase (AmSOD1) mediates hydrogen peroxide tolerance in Avicennia marina

Fig. 5. Effects of incubation at different temperatures and pH on the activity of AmSOD1. The purified AmSOD1 was incubated (A) at temperatures (25, 37, 50, 60 and 75 ◦ C) (B) in buffers with pH (3–12) for 1 h before SOD assay. Then the activity of AmSOD1 was determined as explained in the "Materials and Methods". Each data point represents the mean ± SD obtained from three independent reactions.

Fig. 6 in Comparative transcriptome analysis reveals the regulatory effects of acetylcholine on salt tolerance of Nicotiana benthamiana

Fig. 6. Differentially expressed genes related to the plant hormone signal transduction pathway in the comparison (NaCl vs NaCl + ACh). (A) diagram of auxin, gibberellin, brassinosteroid and salicylic acid signalling transduction pathways; (B) information and expression patterns of differentially expressed genes involved in auxin, gibberellin, brassinosteroid and salicylic acid signalling transduction pathways. Red means upregulated expression of genes, and green means downregulated expression of genes. The number in each sample name represents the sample order. (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 Comparative transcriptome analysis reveals the regulatory effects of acetylcholine on salt tolerance of Nicotiana benthamiana

Fig. 3. Functional annotation of differentially expressed genes (DEGs) based on gene ontology categorization. The left Y-axis represents the significantly enriched GO terms (p <0.05) pathways. The Xaxis represents the percentage of DEGs belonging to the corresponding pathway. The sizes of bubbles represent the number of DEGs in the corresponding pathway, and the colours of the bubbles represent the enrichment p-value of the corresponding pathway. The left y-axis shows the Gene Ontology terms. Biological process, cellular component and molecular function are indicated by different colours. Only significantly enriched GO terms (p <0.05) are shown. (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 Comparative transcriptome analysis reveals the regulatory effects of acetylcholine on salt tolerance of Nicotiana benthamiana

Fig. 8. Quantitative real-time PCR (RT-qPCR) validation of selected differentially expressed genes detected in Nicotiana benthamiana leaves. The expression levels obtained by RT-qPCR are represented in black lines, RT-qPCR data showed the mean values from three replicates, and the error bars represent the SE of the means, while the corresponding expression data for RNA-seq are represented in the white histogram. CN, control; CN + ACh, 10 μM acetylcholine; NaCl, 150 mM NaCl stress; NaCl + ACh, 150 mM NaCl stress plus 10 μM acetylcholine.

Fig. 5 in Comparative transcriptome analysis reveals the regulatory effects of acetylcholine on salt tolerance of Nicotiana benthamiana

Fig. 5. Heatmap representing the differentially expressed genes (DEGs) involved in cell wall extensibility of Nicotiana benthamiana leaves as influenced by NaCl alone or in combination with acetylcholine treatment (NaCl + ACh). Red means upregulated expression of genes, and green means downregulated expression of genes. The number in each sample name represents the sample order. (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 Comparative transcriptome analysis reveals the regulatory effects of acetylcholine on salt tolerance of Nicotiana benthamiana

Fig. 1. Changes in reactive oxygen species accumulation and production and antioxidant enzyme activity in Nicotiana benthamiana leaves 5 days after treatment with acetylcholine (ACh) under salt stress. (a) leaves were stained with NBT and DAB, (b) Fv/Fm, (c) Superoxide content, (d) Hydrogen peroxide content, (e) Ascorbate peroxidase activity and (d) Catalase activity as influenced by salt stress alone or in combination with ACh treatment. Data are means of three replications ±SE. Means with the same lowercase letters are not significantly different at p <0.05, according to Duncan's multiple range test. CN: control; CN + ACh, 10 μM acetylcholine; NaCl, 150 mM NaCl stress; NaCl + ACh, 150 mM NaCl stress plus 10 μM acetylcholine.

Fig. 7 in Comparative transcriptome analysis reveals the regulatory effects of acetylcholine on salt tolerance of Nicotiana benthamiana

Fig. 7. Heatmap representing the differentially expressed genes involved in transcription factors extensibility of Nicotiana benthamiana leaves as influenced by NaCl alone or in combination with acetylcholine treatment (NaCl + ACh). Red means upregulated expression of genes, and green means downregulated expression of genes. The number in each sample name represents the sample order. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4 in Comparative transcriptome analysis reveals the regulatory effects of acetylcholine on salt tolerance of Nicotiana benthamiana

Fig. 4. The Kyoto Encyclopedia of Genes and Genomes pathway enrichment scatter map (p <0.05). The X-axis (Rich Factor) represents the percentage of DEGs belonging to the corresponding pathway.