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Fig. 8 in NtCOMT1 responsible for phytomelatonin biosynthesis confers drought tolerance in Nicotiana tabacum

Fig. 8. The expression level of drought-related genes in NtCOMT1 overexpressing lines, WT, and OE-Empty tobacco after drought treatment. Data are means ±SE of three biological replicates and means followed by different letters are significantly different (p <0.05).

Fig. 7 in NtCOMT1 responsible for phytomelatonin biosynthesis confers drought tolerance in Nicotiana tabacum

Fig. 7. Histochemical staining with DAB for detection of H2O2 (A), the content of H2O2 (B) and MDA (C), and the antioxidant enzyme activities of CAT (D) and SOD (E) in NtCOMT1 overexpressing lines, WT, and OE-Empty tobacco after drought treatment. Data are means ± SE of three biological replicates and means followed by different letters are significantly different (p <0.05).

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. 2 in Zinc uptake and HMA4 activity are required for micro- and macroelement balance in tobacco (Nicotiana tabacum)

Fig. 2. Classification of gene expression changes observed in both the HMA4 double-knockout mutants (cv. #3) and HMA4 RNAi plants (cv.#1) relative to their respective controls. The genes were classified manually according to the putative function of the closest Arabidopsis Blast hit (see Supplementary Table S2).

Fig. 1 in Zinc uptake and HMA4 activity are required for micro- and macroelement balance in tobacco (Nicotiana tabacum)

Fig. 1. (a) Relative element contents of HMA4-impaired plants compared with their controls. HMA4 RNAi plants in cv.#1 background were compared with their WT controls (n = 3 replicates each), and HMA4 double-mutant plants in cv. #3 background were compared with their null-segregant controls (n = 4). The elements were measured in dry weight of lower leaves of plants grown under greenhouse conditions. The error bars depict the 0.95 confidence interval (CI) corrected according to Satterthwaite approximation. (b) Pictures of representative plants of each group (sticker size = 11.9 cm).

Fig. 6 in Zinc uptake and HMA4 activity are required for micro- and macroelement balance in tobacco (Nicotiana tabacum)

Fig. 6. Relative element contents of different HMA4-double mutants versus the null-segregant WT controls in field experiments. The data were consolidated over 4 years of field experiments in Switzerland and 3 years of field experiments in Poland, as described in the Experimental section. The elements were measured in dry weight of leaves (mid-lower leaf position). Bars indicate estimates and confidence intervals (α = 0.1).

Fig. 4 in Zinc uptake and HMA4 activity are required for micro- and macroelement balance in tobacco (Nicotiana tabacum)

Fig. 4. Relative elemental content of N. tabacum cv. #1 and cv. #2 HMA4 RNAi plants compared with their WT and of HMA4 double-knockout mutants compared with their null-segregants, for four different fertilization conditions: (a) normal fertilization, (b) fertilization containing lower concentrations of phosphate, (c) fertilization containing higher concentrations of Zn, and (d) fertilization with low concentrations of phosphate and high concentrations of Zn. The high Zn condition was achieved by addition of 0.1 g additional Zn per plant. The elements were measured in dry weight of leaves (mid-lower leaf position). Absolute data are listed in Supplementary Table S3. Data represent mean (0.95 CI) of n = 5 replicate plants, corrected according to Satterthwaite approximation. (Three outliers were removed for condition (d): Al and Cu for cv.#2; Ti for cv.#3.).

Fig. 3 in Zinc uptake and HMA4 activity are required for micro- and macroelement balance in tobacco (Nicotiana tabacum)

Fig. 3. Working model of metabolic changes in HMA4-impaired plants based on the gene expression changes observed in the roots and leaves of HMA4 RNAi and HMA4-mutant N. tabacum plants relative to their controls. The lack of HMA4 function causes Zn deficiency in leaves, leading to a local Zn-deficiency response. In roots, a Fe-deficiency response is observed, caused either by a high Zn:Fe ratio in the roots or by a systemic Zn-deficiency signal. The Fe-deficiency signal leads—via the transcription factor FIT1—to upregulation of the Fe- and Mn- transporter genes IRT1 and NRAMP1 and also induces the synthesis and export of iron-binding compounds (IBC), thus additionally assisting Fe uptake. At the same time, a P-deficiency signal involving the transcription factor HHO2 is generated in the leaves. In roots, the P-deficiency signal leads to increased expression of genes encoding acid phosphatases, enabling increased phosphate uptake from the rhizosphere. Consequently, Fe, Mn, Cu, and P accumulate in the leaves. P is stored in vacuoles (via PHT5; 1, PHT5; 3) and also exported to mitochondria and chloroplasts. Cu is detoxified by transport to the chloroplasts, which are a major site of Cu use because of Cu-containing plastocyanin production. However, high levels of Cu are toxic for the photosynthetic electron transport system, especially for photosystem II, which is reflected in the upregulation of PII light harvesting complex genes (LHCB). At the same time, Cu uptake to the cells is limited by downregulation of COPT1. Ion uptake might cause a lower osmotic potential, leading to water influx via various aquaporin genes. High turgor pressure or Fe excess ultimately affects the cell wall, as observed by the increased expression of genes involved in cell wall biosynthesis, expansion, and crosslinking. bHLH115: basic helix-loop-helix transcription factor; CA: carbonic anhydrase; CESA: cellulose synthase; COPT: copper transporter; DMRL: dimethyl-8-ribityllumazine synthase; EXPA: expansin; FIT: FER-like regulator of iron uptake (transcription factor); HHO2: hypersensitivity to low phosphate-elicited primary root shortening 1 homolog 2 (myb-like transcription factor); HMA: heavy metal ATPase; IBC: iron binding compound; IRT: iron-regulated transporter; LAC: laccase; LHCB: photosystem II light harvesting complex; NA: nicotianamine; NAS: nicotianamine synthase; NRAMP: natural resistance-associated macrophage protein (metal ion transporter); PDR: pleiotropic drug resistance (ATP-binding cassette transporter); PHT: phosphate transporter; PIP: plasma membrane intrinsic protein; PME: pectin methyl esterase; PMEI: pectin methyl esterase inhibitor; PRX: peroxidase; RBCS: ribulose bisphosphate carboxylase small chain; TIP: tonoplast intrinsic protein; TBL: TRICHOME BIREFRINGENCE-LIKE (xylan acetylation); ZIP: zinc transporter.

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Fig. 1 in Cyclopiazonic acid type indole alkaloids from Nicotiana tabacum-derived fungus Aspergillus versicolor and their anti-tobacco mosaic virus activities

Fig. 1. The structures of compounds 1–8 from A. versicolor.

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Fig. 2 in Cyclopiazonic acid type indole alkaloids from Nicotiana tabacum-derived fungus Aspergillus versicolor and their anti-tobacco mosaic virus activities

Fig. 2. The key HMBC () and 1H–1H COSY () correlations of compounds 1 and 3.

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openGEO-OpenNov 2019View details →

Nicotiana tabacum degradome analysis using high definition adapters proves an efficient targeting method against the Tomato leaf curl New Delhi virus using artificial ta-siRNAs

GEO Series GSE85819. Nicotiana tabacum. 6 samples. Type: Non-coding RNA profiling by high throughput sequencing; Expression profiling by high throughput sequencing.

openGEO-OpenJan 2017View details →

Integrated transcriptomic and metabolomic to characterize cold stress response in Nicotiana tabacum

GEO Series GSE94105. Nicotiana tabacum. 8 samples. Type: Expression profiling by high throughput sequencing.

openGEO-OpenJul 2019View details →

Transcriptome anaylsis of wild type 2-celled proembryo of Nicotiana tabacum

GEO Series GSE133373. Nicotiana tabacum. 3 samples. Type: Expression profiling by high throughput sequencing.

openGEO-OpenJan 2024View details →

Transcriptome analysis reveals the key role of overdominant expression of photosynthetic and respiration-related genes in the formation of tobacco(Nicotiana tabacum L.) biomass heterosis

GEO Series GSE252097. Nicotiana tabacum. 18 samples. Type: Expression profiling by high throughput sequencing.

openGEO-OpenJun 2021View details →

Transcriptome analysis of heat stressed Nicotiana tabacum pollen tubes

GEO Series GSE153474. Nicotiana tabacum. 9 samples. Type: Expression profiling by high throughput sequencing.