Data from: Nuclear-localized β-(1→3)-glucan polymers and expression of the candidate metabolic genes provide evidence of a novel photosynthate storage system in Paulinella micropora
Data files
Mar 03, 2026 version files 57.01 MB
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GH161.aln.fasta
1.68 MB
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GH161.aln.trim.fasta
258.87 KB
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GH161.pdf
427.20 KB
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GH161.treefile
35.52 KB
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GH72.aln.fasta
2.73 MB
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GH72.aln.trim.fasta
80.29 KB
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GH72.pdf
417.38 KB
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GH72.treefile
35.39 KB
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GH81.aln.fasta
18.25 MB
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GH81.aln.trim.fasta
274.40 KB
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GH81.pdf
665.49 KB
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GH81.treefile
83.15 KB
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GT48.aln.fasta
30.99 MB
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GT48.aln.trim.fasta
555.52 KB
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GT48.pdf
450.64 KB
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GT48.treefile
75.64 KB
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README.md
5.32 KB
Abstract
Paulinella micropora is a testate amoeba that acquired a photosynthetic organelle (the chromatophore) from a cyanobacterial donor. This event is independent from the origin of the widespread primary plastid in Archaeplastida. Whereas photosynthate storage mechanisms are well-characterized in plants, little is known about how P. micropora stores and utilizes photosynthetically fixed carbon. Here, we examined β-glucans in P. micropora, focusing on their identification, subcellular localization, and associated gene expression patterns to assess their putative roles in the cell. Comparative analyses suggest that photosynthetic Paulinella species lack the genes required for α-glucan (starch or glycogen) storage but contain β-glucans, as confirmed by enzymatic hydrolysis in P. micropora that released glucose monomers. Using immunolabeling transmission electron microscopy with a monoclonal anti-(1→3)-β-D-glucan antibody, we report that β-1,3-glucan is localized to the chromatophore and cytosol, and unexpectedly also to the nucleus, where labeling was preferentially enriched in heterochromatin. The latter result has not previously been reported for β-glucans. In addition, co-expression analysis of transcriptome data identified candidate genes likely involved in β-glucan metabolism in Paulinella, several of which show signatures of horizontal gene transfer, suggesting a mosaic evolutionary origin. Our results provide novel insights into the evolution of photosynthate storage strategies and the potential roles of β-glucans in an independent example of primary plastid endosymbiosis.
Dataset DOI: 10.5061/dryad.mcvdnckfh
Description of the data and file structure
Phylogenetic analyses were performed for candidate β-glucan metabolism genes identified in P. micropora. Eukaryotic homologs were retrieved from the EukProt v3 database and clustered at 90 % sequence identity using CD-HIT to reduce redundancy. Bacterial and archaeal homologs were obtained from the GTDB-Tk v2 and downsampled at a 70 % identity threshold using CD-HIT. Viral sequences were sourced from the Virus–Host DB and clustered at 90 % identity using CD-HIT. In addition, a heterotrophic Paulinella MAG was included in the analysis.
Using P. micropora sequences as queries, homology searches were conducted using DIAMOND v 2.1.2 in ultra-sensitive mode with an e-value cutoff of 0.001. To minimize biases arising from uneven taxonomic representation across databases, searches were performed separately against seven taxonomically partitioned databases: Prokaryotes, Archaeplastida, Amorphea, Stramenopiles, Alveolata, other eukaryotes, and viruses. Sequences within each group were aligned using MAFFT v7.490 with the L-INS-i algorithm. Poorly aligned sequences and partial fragments were manually inspected and removed. The remaining sequences were realigned using MAFFT (L-INS-i), followed by automated trimming with trimAl to remove ambiguously aligned regions. Phylogenetic trees were inferred using IQ-TREE 2.0.7 with automatic model selection (ModelFinder), and node support was evaluated using 1,000 ultrafast bootstrap and 1,000 SH-aLRT replicates.
Files and variables
File: GH81.aln.trim.fasta
Description: Trimmed multiple sequence alignment of GH81 homologous protein sequences used for phylogenetic analysis.
File: GH81.treefile
Description: Maximum-likelihood phylogenetic tree of GH81 homologs inferred using IQ-TREE2.
File: GH81.aln.fasta
Description: Untrimmed multiple sequence alignment of GH81 homologous protein sequences generated using MAFFT.
File: GT48.aln.trim.fasta
Description: Trimmed multiple sequence alignment of GT48 homologous protein sequences used for phylogenetic analysis.
File: GT48.treefile
Description: Maximum-likelihood phylogenetic tree of GT48 homologs inferred using IQ-TREE2.
File: GH72.aln.trim.fasta
Description: Trimmed multiple sequence alignment of GH72 homologous protein sequences used for phylogenetic analysis.
File: GH72.treefile
Description: Maximum-likelihood phylogenetic tree of GH72 homologs inferred using IQ-TREE2.
File: GH161.aln.trim.fasta
Description: Trimmed multiple sequence alignment of GH161 homologous protein sequences used for phylogenetic analysis.
File: GH161.treefile
Description: Maximum-likelihood phylogenetic tree of GH161 homologs inferred using IQ-TREE2.
File: GH72.aln.fasta
Description: Untrimmed multiple sequence alignment of GH72 homologous protein sequences generated using MAFFT.
File: GH161.aln.fasta
Description: Untrimmed multiple sequence alignment of GH161 homologous protein sequences generated using MAFFT.
File: GT48.aln.fasta
Description: Untrimmed multiple sequence alignment of GT48 homologous protein sequences generated using MAFFT.
File: GT48.pdf
Description: Phylogeny of GT48 homologs. Paulinella sequences are shown in blue; pink, Rhizaria; orange, Stramenopiles; olive, Alveolata; green, Archaeplastida. Major clades are collapsed for clarity. Only ultrafast bootstrap values ≥80, representing well-supported nodes, are shown. P. micropora sequences belonging to the co-expression modules m10 and m13 are indicated next to the corresponding branches.
File: GH72.pdf
Description: Phylogeny of GH72 homologs. Paulinella sequences are shown in blue; pink, Rhizaria; orange, Stramenopiles; olive, Alveolata; green, Archaeplastida. Major clades are collapsed for clarity. Only ultrafast bootstrap values ≥ 80, representing well-supported nodes, are shown. P. micropora sequences belonging to the co-expression modules m10 and m13 are indicated next to the corresponding branches.
File: GH161.pdf
Description: Phylogeny of GH161 homologs. Paulinella sequences are shown in blue; pink, Rhizaria; orange, Stramenopiles; olive, Alveolata; green, Archaeplastida. Major clades are collapsed for clarity. Only ultrafast bootstrap values ≥ 80, representing well-supported nodes, are shown. P. micropora sequences belonging to the co-expression modules m10 and m13 are indicated next to the corresponding branches.
File: GH81.pdf
Description: Phylogeny of GH81 homologs. Paulinella sequences are shown in blue; pink, Rhizaria; orange, Stramenopiles; olive, Alveolata; green, Archaeplastida. Major clades are collapsed for clarity. Only ultrafast bootstrap values ≥80, representing well-supported nodes, are shown. P. micropora sequences belonging to the co-expression modules m10 and m13 are indicated next to the corresponding branches.
