Review




Structured Review

BGI Genomics Co sequencing data
CHH methylation sites, particularly those with high-methylation levels (≥90%), are rare in plants. This figure presents the statistics on high-methylation CHH sites from previously published bisulfite <t>sequencing</t> (BS-seq) datasets , – and those generated in this study. a Ratios of high-methylation CHH sites among quantified CHH motifs (≥5 read coverage) (left panel) and the number of covered 9-mer contexts (right panel) in BS-seq datasets from ten plant species. Only 9-mers observed at three or more high-methylation CHH sites were considered. Species other than A. thaliana and O. sativa were selected based on high CHH methylation ratios reported in previous studies , . b Ratios of high-methylation CHH sites in BS-seq datasets sequenced for this study, including A. thaliana , O. sativa , and species with abundant high-methylation CHH sites identified in ( a ), as well as Glycine max and Marchantia polymorpha . c Number of covered 9-mer contexts (top) and heatmap of context abundance (bottom) grouped by CHH motifs in nanopore datasets from six plant species. In the top panel, the top line of each bar corresponds to the number (36,864) of all possible 9-mer sequences centered with a CHH motif. A 9-mer was considered covered if present in 50 or more positive training samples and had at least an equal number of negative samples. “Mixed” refers to combined samples from S. miltiorrhiza , R. communis , and S. tuberosum . Source data are provided as a Source Data file.
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1) Product Images from "Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant"

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant

Journal: Nature Communications

doi: 10.1038/s41467-025-58576-x

CHH methylation sites, particularly those with high-methylation levels (≥90%), are rare in plants. This figure presents the statistics on high-methylation CHH sites from previously published bisulfite sequencing (BS-seq) datasets , – and those generated in this study. a Ratios of high-methylation CHH sites among quantified CHH motifs (≥5 read coverage) (left panel) and the number of covered 9-mer contexts (right panel) in BS-seq datasets from ten plant species. Only 9-mers observed at three or more high-methylation CHH sites were considered. Species other than A. thaliana and O. sativa were selected based on high CHH methylation ratios reported in previous studies , . b Ratios of high-methylation CHH sites in BS-seq datasets sequenced for this study, including A. thaliana , O. sativa , and species with abundant high-methylation CHH sites identified in ( a ), as well as Glycine max and Marchantia polymorpha . c Number of covered 9-mer contexts (top) and heatmap of context abundance (bottom) grouped by CHH motifs in nanopore datasets from six plant species. In the top panel, the top line of each bar corresponds to the number (36,864) of all possible 9-mer sequences centered with a CHH motif. A 9-mer was considered covered if present in 50 or more positive training samples and had at least an equal number of negative samples. “Mixed” refers to combined samples from S. miltiorrhiza , R. communis , and S. tuberosum . Source data are provided as a Source Data file.
Figure Legend Snippet: CHH methylation sites, particularly those with high-methylation levels (≥90%), are rare in plants. This figure presents the statistics on high-methylation CHH sites from previously published bisulfite sequencing (BS-seq) datasets , – and those generated in this study. a Ratios of high-methylation CHH sites among quantified CHH motifs (≥5 read coverage) (left panel) and the number of covered 9-mer contexts (right panel) in BS-seq datasets from ten plant species. Only 9-mers observed at three or more high-methylation CHH sites were considered. Species other than A. thaliana and O. sativa were selected based on high CHH methylation ratios reported in previous studies , . b Ratios of high-methylation CHH sites in BS-seq datasets sequenced for this study, including A. thaliana , O. sativa , and species with abundant high-methylation CHH sites identified in ( a ), as well as Glycine max and Marchantia polymorpha . c Number of covered 9-mer contexts (top) and heatmap of context abundance (bottom) grouped by CHH motifs in nanopore datasets from six plant species. In the top panel, the top line of each bar corresponds to the number (36,864) of all possible 9-mer sequences centered with a CHH motif. A 9-mer was considered covered if present in 50 or more positive training samples and had at least an equal number of negative samples. “Mixed” refers to combined samples from S. miltiorrhiza , R. communis , and S. tuberosum . Source data are provided as a Source Data file.

Techniques Used: Methylation, Methylation Sequencing, Generated

Quantitative evaluation of CHH methylation detection by DeepPlant and Dorado
Figure Legend Snippet: Quantitative evaluation of CHH methylation detection by DeepPlant and Dorado

Techniques Used: Methylation, Sequencing

a Circos plot illustrating DeepPlant CHH methylation profiling in centromeric regions and 100 kb intervals upstream and downstream in O. sativa . From outer to inner: ideograms of centromere (center box) and neighboring regions (two terminal boxes); histograms of normalized sequencing coverage across 100 bp bins (gray, normalized against mean genomic coverage); histograms of CHH methylation frequencies (blue) across 100 bp bins. b Comparison of CHH motif coverage ratios across different genomic regions between BS-seq, DeepPlant, and Dorado profiling. To be noticed, the same nanopore dataset was used for DeepPlant and Dorado profiling, and the coverage difference between DeepPlant and Dorado derived from the distinct read filters they applied. DeepPlant applies three thresholds for screening high-quality alignments, including MAPQ ≥ 20, primary alignment length/read length ≥80%, and mapping identity ≥80% by default. Only CHH motifs with a minimum read coverage of 10 were regarded as quantified. TE transposable element, CDS non-TE protein-coding sequences. c Read coverage and CHH methylation frequencies in the centromeric regions of Chr1, comparing whole-genome BS-seq data, DeepPlant, and Dorado analysis on Nanopore data. d Boxplot illustrating CHH methylation frequencies on the forward (+) and reverse (−) strands in protein-coding and transposable element (TE) regions. The center line represents the median; each box shows the first and third quartiles; minima represents the larger between Q1−1.5×IQR and the minimum observed value; maxima represents the smaller between Q3 + 1.5×IQR and the maximum observed value. e Violin plot displaying strand-specific CHH methylation status across various TE types and non-TE protein-coding regions. The annotation of TEs and protein-coding regions was acquired from T2T-NIP . Source data of ( c , d ) are provided in Zenodo [10.5281/zenodo.15062213]. Source data of the other panels are provided as a Source Data file.
Figure Legend Snippet: a Circos plot illustrating DeepPlant CHH methylation profiling in centromeric regions and 100 kb intervals upstream and downstream in O. sativa . From outer to inner: ideograms of centromere (center box) and neighboring regions (two terminal boxes); histograms of normalized sequencing coverage across 100 bp bins (gray, normalized against mean genomic coverage); histograms of CHH methylation frequencies (blue) across 100 bp bins. b Comparison of CHH motif coverage ratios across different genomic regions between BS-seq, DeepPlant, and Dorado profiling. To be noticed, the same nanopore dataset was used for DeepPlant and Dorado profiling, and the coverage difference between DeepPlant and Dorado derived from the distinct read filters they applied. DeepPlant applies three thresholds for screening high-quality alignments, including MAPQ ≥ 20, primary alignment length/read length ≥80%, and mapping identity ≥80% by default. Only CHH motifs with a minimum read coverage of 10 were regarded as quantified. TE transposable element, CDS non-TE protein-coding sequences. c Read coverage and CHH methylation frequencies in the centromeric regions of Chr1, comparing whole-genome BS-seq data, DeepPlant, and Dorado analysis on Nanopore data. d Boxplot illustrating CHH methylation frequencies on the forward (+) and reverse (−) strands in protein-coding and transposable element (TE) regions. The center line represents the median; each box shows the first and third quartiles; minima represents the larger between Q1−1.5×IQR and the minimum observed value; maxima represents the smaller between Q3 + 1.5×IQR and the maximum observed value. e Violin plot displaying strand-specific CHH methylation status across various TE types and non-TE protein-coding regions. The annotation of TEs and protein-coding regions was acquired from T2T-NIP . Source data of ( c , d ) are provided in Zenodo [10.5281/zenodo.15062213]. Source data of the other panels are provided as a Source Data file.

Techniques Used: Methylation, Sequencing, Comparison, Derivative Assay

Related Articles

Polymerase Chain Reaction:

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant.
Article Snippet: .. After bisulfite conversion, PCR amplificationwas performed, and the sequencing data were generated by BGI Genomics Co. Ltd. ..

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant
Article Snippet: .. After bisulfite conversion, PCR amplification was performed, and the sequencing data were generated by BGI Genomics Co. Ltd. ..

Sequencing:

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant.
Article Snippet: .. After bisulfite conversion, PCR amplificationwas performed, and the sequencing data were generated by BGI Genomics Co. Ltd. ..

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant
Article Snippet: .. After bisulfite conversion, PCR amplification was performed, and the sequencing data were generated by BGI Genomics Co. Ltd. ..

Generated:

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant.
Article Snippet: .. After bisulfite conversion, PCR amplificationwas performed, and the sequencing data were generated by BGI Genomics Co. Ltd. ..

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant
Article Snippet: .. After bisulfite conversion, PCR amplification was performed, and the sequencing data were generated by BGI Genomics Co. Ltd. ..

Amplification:

Article Title: Accurate cross-species 5mC detection for Oxford Nanopore sequencing in plants with DeepPlant
Article Snippet: .. After bisulfite conversion, PCR amplification was performed, and the sequencing data were generated by BGI Genomics Co. Ltd. ..



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Image Search Results


Strains and plasmids used in this study.

Journal: Frontiers in Microbiology

Article Title: Thiamine-Mediated Cooperation Between Auxotrophic Rhodococcus ruber ZM07 and Escherichia coli K12 Drives Efficient Tetrahydrofuran Degradation

doi: 10.3389/fmicb.2020.594052

Figure Lengend Snippet: Strains and plasmids used in this study.

Article Snippet: The draft genome sequence data of strain ZM07 have been included in the National Center for Biotechnology Information (NCBI), accession number JACVXT000000000 , and strain K12 genome sequence data can also be found in NCBI (GenBank accession number NC_000913.3 ).

Techniques: Mutagenesis, Plasmid Preparation, Expressing

THF degradation and growth curves of strain ZM07 monocultured (A) and cocultured with strain K12 (B) in the first, second and third transfers. The solid lines represent THF concentration curves; the dotted lines represent growth curves (OD 600 ); the circles represent the first transfer; the squares represent the second transfer; and the triangles represent the third transfer. Error bars: SD from three independent replicates, which may be smaller than the marker.

Journal: Frontiers in Microbiology

Article Title: Thiamine-Mediated Cooperation Between Auxotrophic Rhodococcus ruber ZM07 and Escherichia coli K12 Drives Efficient Tetrahydrofuran Degradation

doi: 10.3389/fmicb.2020.594052

Figure Lengend Snippet: THF degradation and growth curves of strain ZM07 monocultured (A) and cocultured with strain K12 (B) in the first, second and third transfers. The solid lines represent THF concentration curves; the dotted lines represent growth curves (OD 600 ); the circles represent the first transfer; the squares represent the second transfer; and the triangles represent the third transfer. Error bars: SD from three independent replicates, which may be smaller than the marker.

Article Snippet: The draft genome sequence data of strain ZM07 have been included in the National Center for Biotechnology Information (NCBI), accession number JACVXT000000000 , and strain K12 genome sequence data can also be found in NCBI (GenBank accession number NC_000913.3 ).

Techniques: Concentration Assay, Marker

Comparison of the KEGG metabolic pathways reconstructed from the complete genomes of ZM07 and K12. Here, thiamine metabolism (map00730) is shown. The ZM07-specific pathways are colored in green; the K12-specific pathways are colored in blue; shared pathways are colored in brown; and pathways present in neither ZM07 nor K12 are colored in white. The genes for thiF (No. 2.7.7.73), thiH (No. 4.1.99.19), and thiI (No. 2.8.1.4) are highlighted by purple boxes, and the gene for thiE (No. 2.5.1.3) is highlighted by red boxes.

Journal: Frontiers in Microbiology

Article Title: Thiamine-Mediated Cooperation Between Auxotrophic Rhodococcus ruber ZM07 and Escherichia coli K12 Drives Efficient Tetrahydrofuran Degradation

doi: 10.3389/fmicb.2020.594052

Figure Lengend Snippet: Comparison of the KEGG metabolic pathways reconstructed from the complete genomes of ZM07 and K12. Here, thiamine metabolism (map00730) is shown. The ZM07-specific pathways are colored in green; the K12-specific pathways are colored in blue; shared pathways are colored in brown; and pathways present in neither ZM07 nor K12 are colored in white. The genes for thiF (No. 2.7.7.73), thiH (No. 4.1.99.19), and thiI (No. 2.8.1.4) are highlighted by purple boxes, and the gene for thiE (No. 2.5.1.3) is highlighted by red boxes.

Article Snippet: The draft genome sequence data of strain ZM07 have been included in the National Center for Biotechnology Information (NCBI), accession number JACVXT000000000 , and strain K12 genome sequence data can also be found in NCBI (GenBank accession number NC_000913.3 ).

Techniques: Comparison

(A) Growth curves of strain ZM07 monocultured (pink line) and cocultured with K12 under the non-contact condition (blue line) in two-phase reactors and the growth curve of the two strains cocultured under the contact condition (yellow line). (B) THF degradation curves of strain ZM07 in the three cases described above. (C) Growth curve of strain K12 cocultured with ZM07 under the non-contact condition in a two-phase reactor. (D) Two-phase reactor used to separate the cultured strains in this study. The P -value indicates a significant difference between monocultured strain ZM07 and strain ZM07 cocultured with K12 in two-phase reactors and was determined using Student’s t -test ( n = 3, ∗∗∗ p < 0.001). Error bars: SD from three independent replicates, which may be smaller than the marker.

Journal: Frontiers in Microbiology

Article Title: Thiamine-Mediated Cooperation Between Auxotrophic Rhodococcus ruber ZM07 and Escherichia coli K12 Drives Efficient Tetrahydrofuran Degradation

doi: 10.3389/fmicb.2020.594052

Figure Lengend Snippet: (A) Growth curves of strain ZM07 monocultured (pink line) and cocultured with K12 under the non-contact condition (blue line) in two-phase reactors and the growth curve of the two strains cocultured under the contact condition (yellow line). (B) THF degradation curves of strain ZM07 in the three cases described above. (C) Growth curve of strain K12 cocultured with ZM07 under the non-contact condition in a two-phase reactor. (D) Two-phase reactor used to separate the cultured strains in this study. The P -value indicates a significant difference between monocultured strain ZM07 and strain ZM07 cocultured with K12 in two-phase reactors and was determined using Student’s t -test ( n = 3, ∗∗∗ p < 0.001). Error bars: SD from three independent replicates, which may be smaller than the marker.

Article Snippet: The draft genome sequence data of strain ZM07 have been included in the National Center for Biotechnology Information (NCBI), accession number JACVXT000000000 , and strain K12 genome sequence data can also be found in NCBI (GenBank accession number NC_000913.3 ).

Techniques: Cell Culture, Marker

Relative abundances of ZM07, K12, and K12Δ thiE in a two-strain system (ZM07 and K12) (A) and in a three-strain system (B) across different transfers without thiamine. Relative abundances of three strains in a two-strain system (ZM07 and trace amount of K12) (C) and in a three-strain system (ZM07, K12 and trace amount of K12Δ thiE ) (D) across different transfers. The plots within (B,D) show details of the relative abundance of K12Δ thiE in corresponding experiments. Three independent replicates were performed for each group. Error bars: SD from three independent replicates.

Journal: Frontiers in Microbiology

Article Title: Thiamine-Mediated Cooperation Between Auxotrophic Rhodococcus ruber ZM07 and Escherichia coli K12 Drives Efficient Tetrahydrofuran Degradation

doi: 10.3389/fmicb.2020.594052

Figure Lengend Snippet: Relative abundances of ZM07, K12, and K12Δ thiE in a two-strain system (ZM07 and K12) (A) and in a three-strain system (B) across different transfers without thiamine. Relative abundances of three strains in a two-strain system (ZM07 and trace amount of K12) (C) and in a three-strain system (ZM07, K12 and trace amount of K12Δ thiE ) (D) across different transfers. The plots within (B,D) show details of the relative abundance of K12Δ thiE in corresponding experiments. Three independent replicates were performed for each group. Error bars: SD from three independent replicates.

Article Snippet: The draft genome sequence data of strain ZM07 have been included in the National Center for Biotechnology Information (NCBI), accession number JACVXT000000000 , and strain K12 genome sequence data can also be found in NCBI (GenBank accession number NC_000913.3 ).

Techniques:

Relative abundances of ZM07, K12, and K12Δ thiE in two-strain systems (ZM07 and K12) (A) , (ZM07 and K12Δ thiE ) (B) and a three-strain system (C) in different transfers with 0.01 mM thiamine. (D) Relative abundances of K12 and K12Δ thiE in the above experiments in two-strain systems (the relative abundance of strain K12 in a cocultured system of ZM07 and K12 in (A) and the relative abundance of strain K12Δ thiE in a cocultured system of ZM07 and K12Δ thiE in panel (B) in different transfers with 0.01 mM thiamine. (E) Relative abundances of K12 and K12Δ thiE in the above experiments in the three-strain system (ZM07, K12, and K12Δ thiE in (C) in different transfers with 0.01 mM thiamine. Three independent replicates were performed for each group. Error bars: SD from three independent replicates. The P value indicates statistical significance between K12 and K12Δ thiE determined using Student’s t -test ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Journal: Frontiers in Microbiology

Article Title: Thiamine-Mediated Cooperation Between Auxotrophic Rhodococcus ruber ZM07 and Escherichia coli K12 Drives Efficient Tetrahydrofuran Degradation

doi: 10.3389/fmicb.2020.594052

Figure Lengend Snippet: Relative abundances of ZM07, K12, and K12Δ thiE in two-strain systems (ZM07 and K12) (A) , (ZM07 and K12Δ thiE ) (B) and a three-strain system (C) in different transfers with 0.01 mM thiamine. (D) Relative abundances of K12 and K12Δ thiE in the above experiments in two-strain systems (the relative abundance of strain K12 in a cocultured system of ZM07 and K12 in (A) and the relative abundance of strain K12Δ thiE in a cocultured system of ZM07 and K12Δ thiE in panel (B) in different transfers with 0.01 mM thiamine. (E) Relative abundances of K12 and K12Δ thiE in the above experiments in the three-strain system (ZM07, K12, and K12Δ thiE in (C) in different transfers with 0.01 mM thiamine. Three independent replicates were performed for each group. Error bars: SD from three independent replicates. The P value indicates statistical significance between K12 and K12Δ thiE determined using Student’s t -test ( n = 3, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: The draft genome sequence data of strain ZM07 have been included in the National Center for Biotechnology Information (NCBI), accession number JACVXT000000000 , and strain K12 genome sequence data can also be found in NCBI (GenBank accession number NC_000913.3 ).

Techniques:

Proposed schematic depiction of the interaction mechanism among strains ZM07, K12, and K12Δ thiE during THF degradation. Strain K12 provides thiamine (red circles) to ZM07 and K12Δ thiE , and strain ZM07 might provide succinate or other downstream products (purple diamonds) to K12 and K12Δ thiE as a carbon source by degrading THF. In thiamine-free medium, ZM07 and K12 formed a stable symbiotic system, and K12Δ thiE disappeared during passaging; in thiamine-rich media, both K12 and K12Δ thiE represented only a small proportion of the coculture system.

Journal: Frontiers in Microbiology

Article Title: Thiamine-Mediated Cooperation Between Auxotrophic Rhodococcus ruber ZM07 and Escherichia coli K12 Drives Efficient Tetrahydrofuran Degradation

doi: 10.3389/fmicb.2020.594052

Figure Lengend Snippet: Proposed schematic depiction of the interaction mechanism among strains ZM07, K12, and K12Δ thiE during THF degradation. Strain K12 provides thiamine (red circles) to ZM07 and K12Δ thiE , and strain ZM07 might provide succinate or other downstream products (purple diamonds) to K12 and K12Δ thiE as a carbon source by degrading THF. In thiamine-free medium, ZM07 and K12 formed a stable symbiotic system, and K12Δ thiE disappeared during passaging; in thiamine-rich media, both K12 and K12Δ thiE represented only a small proportion of the coculture system.

Article Snippet: The draft genome sequence data of strain ZM07 have been included in the National Center for Biotechnology Information (NCBI), accession number JACVXT000000000 , and strain K12 genome sequence data can also be found in NCBI (GenBank accession number NC_000913.3 ).

Techniques: Passaging