This project folder contains working data for bioinformatic and functional analyses of diatom chloroplast glycolysis, chloroplast-targeted transporters, and ATP synthases. Feel free to add in contributions you are making here, although please: - update the wiki page with a brief description (name, date, where to look, brief description) - attach a README file (methodology, key results, take home messages) with your data upload. **CONTENTS** **Glycolysis** *Reference phylogenies* (Richard Dorrell, 03/01/21) contains accessions, alignments and consensus RAxML tree topologies for organelle-targeted copies of enolase, and forms 1 and 2 phospho-glycerate mutase, from cryptomonad, haptophyte and ochrophyte genomes and transcriptomes. *Tara Oceans calculations* (Richard Dorrell, Juan José Pierella Karlusich, 03/01/21) contains accessions, alignments, RAxML tree topologies, and relative abundance calculations for sequences phylogenetically reconciled to plastid- or mitochondrial-targeted copies of diatom enolase and PGAM sequences; across all stations and size fractions in *Tara* Oceans metaG and metaT data. *Mutant genotypes and qPCR* (Richard Dorrell, Tomomi Nonoyama, Amandine Baylet, 03/01/21) contains sequences, and qRT-PCR expression profiles for Phaeodactylum colonies in which gene sequences of plastidial enolase (Pt_41515), PGAM1A (Pt_17086) and PGAM2 (Pt_37201) were disrupted by CRISPR-CAS9 knockout. *Protein localisations* (Richard Dorrell, Pauline Clémente, Shun-Min Yang, 25/08/22) provides confocal microscopy images of ptEnolase, ptPGAM1A and ptPGAM2:GFP overexpressor lines, either produced from the transformation of wild-type cells or complementation of zeocin-resistant knockout lines with blasticidin-resistant constructs, confirming in each case a probable chloroplastic localisation. *Growth measurements* (Richard Dorrell, Quentin Caris, Sandrine Adiba, 25/08/22) contains raw growth curves (as measured by flow cytometry), and also calibration curves to measured densities with Malassez cells, for glycolysis knockout, control and complemented lines under multiple conditions, notably varying temperature (19C v 8C) and illumination regime (12h:12h LD cycling, versus 24h continuous light). *Transcriptional dynamics* (Richard Dorrell, Yonna Lauruol, 21/01/21) presents the relative expression of enolase and phospho-glycerate mutase genes in the *Phaeodactylum* genome in previously published meta-analyses of microarray (Ashworth et al., 2016) and RNAseq data (Ait-Mohamed et al., 2020); and in unpublished RNAseq data for control lines generated in this study. Correlation coefficients are additionally presented for the mutated genes (Pt_41515, Pt_17086, Pt_37201) against all genes in the *Phaeodactylum* genome; and against previously completed metabolic maps (Ait-Mohamed et al. 2020) for the *Phaeodactylum* chloroplast and mitochondria. *RNAseq* (Richard Dorrell, Yue Liang, Mathias Penot, 25/08/22) presents crude and edited RNAseq analyses, alongside qPCR validations, for control and glycolysis mutant lines grown under 19C LD, 19C CL, and 8C CL conditions. *Lipidomic analyses* (Richard Dorrell, Nolwenn Guéguen, Valérie Gros, Juliette Jouhet, Eric Maréchal, 25/08/22) presents GC-MS and LC-MS fatty acid profiles of control and glycolysis mutant lines grown under 19C LD, 19C CL, and 8C CL conditions. *Metabolomic analyses* (Richard Dorrell, Youjun Zhang, Alisdair Fernie, 25/08/2022) presents GC-MS analyses of 39 soluble sugars and amino acids in control and glycolysis mutant lines grown under 19C LD, 19C CL, and 8C CL conditions. *Photophysiology* (Richard Dorrell, Dany Croteau, Benjamin Bailleul, 25/08/2022) presents ETR, NPQ and FIRE measurements of control, glycolysis mutant, and complemented glycolysis knockout lines grown under 19C LD and 19C CL. *Paper figures* (Richard Dorrell, Chris Bowler, 25/08/22) presents a provisional manuscript text, main text and supporting figures and tables as uploaded to *BioRXiv*. **Transporters** (Shun Liu, 25/08/22) presents... *Raw TARA alignments* (Shun Liu 04/21) Mafft output file of each nferred plastid transporter raw alignment result with raw diatom plastid transporter homologues from TARA Ocean dataset combined with reference dataset from tree of life *Final alignment and NJ tree for TARA* (Shun Liu 04/21) Curated alignments and tree topologies after manually remove poorly aligned sequences from Raw TARA alignments, defined visually by <50% overlap with the conserved domain region of the query transporter, sequences clade together with diatoms were remained. *Meta G abundance matrix final* (Shun Liu 04/21) Meta-Genome abundance of P. tricornutum plastid transporters final Tara homologous sequences to environmental factors from Tara Oceans Version 1 dataset on the Ocean Gene Atlas website. *Meta T abundance matrix final* (Shun Liu 04/21) Meta-Transcriptome abundance of P. tricornutum plastid transporters final Tara homologous sequences to environmental factors from Tara Oceans Version 1 dataset on the Ocean Gene Atlas website. *transporter TARA homologue distribution* (Shun Liu 04/21) Details of P. tricornutum plastid transporters Tara homologue sequences, taxonomic annotations inferred by Ocean Gene Atlas website **xATPa**