BUSCADOR RECOLECTA

Repository containing derived data for the manuscript 'Global collision-risk hotspots of marine traffic and the world's largest fish, the whale shark'., Peer reviewed

Tabla de datos, Peer reviewed

Predicted genes corresponding to the four most common enzymes present in photosynthetic organisms: NADH:ubiquinone reductase (H+-translocating), N-acetyl-gamma-glutamyl-phosphate reductase, DNA-directed RNA polymerase and non-specific serine/threonine protein kinase of 174 metagenomes sequenced during the Malaspina 2010 global expedition., Peer reviewed

1 table., S2 Table. NAAIRS mutants of SAUR63 and root tortuosity index., Peer reviewed

1 table., S3 Table. Primers used., Peer reviewed

1 table., S4 Table. Clones for transformation., Peer reviewed

1 table., S5 Table. Antibodies., Peer reviewed

A) Genomic map showing positions of genes and locations of mutations from CRISPR/Cas9 mutagenesis in the 9x-saur mutant based on the TAIR10 Arabidopsis genome annotation. The first sgRNA directed cuts in both SAUR61 and SAUR64 (green arrows), creating a deletion between them (green bar) and leaving behind a hybrid gene with a frameshift at the junction site (symbolized by a green X). The second sgRNA directed cuts in the remaining genes (blue arrows, with lighter blue indicating slight mismatches between the sgRNA and the genome), leading to deletions (blue bars) and/or frameshift mutations (blue X’s). SAUR gene names are abbreviated as S61 etc. SAUR61-SAUR68 are on chromosome 1 and SAUR75 is on chromosome 5. B,C) 5-day-old seedlings grown on 1x MS/1% Suc medium in long days. Scale bar, 1 mm. D) Hypocotyl lengths of seedlings grown for 4d in short days on 0.5x MS medium. n, 27 (wild type), 22 (9x-saur). E) Cotyledon area of seedlings grown on vertically oriented plates for 6d on MS/1% Suc medium. n, 16 (wild type), 22 (9x-saur). Graphs show means ± s.d. No statistical differences were detected between wild-type and 9x-saur mutant measurements by t-test. F) Sequences of guide RNAs used for CRISPR/Cas9-mediated mutagenesis, wild-type genes, and mutant alleles present in the 9x-saur mutant. Underlines indicate PAM motif adjacent to guide RNA target site, and any mismatches to the guide RNA sequence. Uppercase bold letters indicate insertion mutations. All alleles create frameshift mutations except for saur75-1, which has an in-frame deletion of 13 amino acids in the SAUR domain., Peer reviewed

A-F) Each point represents the length (A, C, E) or width (B, D, F) of a single epidermal cell against distance of that cell from the base of the hypocotyl (x-axis). Seedlings were grown in short days for 2 (A, B), 3 (C, D) or 4 (E, F) days. Cells were measured from confocal z-stack projections using ML1:RFP PM-localized fluorescence (examples in S3G, S3H and S3I Fig). Data are from 3 cell files per seedling, for two seedlings per time point, except that wild type at 4 days and ost2-2 at 3 days data are from just one seedling each. In a replicate experiment with 2-day-old seedlings only, P35S:SAUR63:YFP:HA hypocotyls also had a greater proportion of longer cells than did wild-type hypocotyls., Peer reviewed

A,B) Hypocotyl length (A) and hypocotyl tortuosity index [B, 1 –(distance between ends)/(contour length)]) of seedlings of indicated genotypes grown for 3d in darkness on plates with 0.5x MS medium. C,D) Root length (C) and root tortuosity index (D) of seedlings of indicated genotypes grown for 4d in long days on plates with 1x MS 1% Sucrose medium. E,F) Root length (E) and root tortuosity index (F) of seedlings of indicated genotypes grown for 4d in short days on plates with 0.5x MS medium. Graphs show means ± s.d. Letters in graphs indicate statistical classes based on Tukey’s Honestly Significant Difference test. n from left to right: Panels A,B: 51, 24, 26, 28, 23, 25, 25; Panels C,D: 34, 17, 23, 14, 19, 15, 16; Panels E,F: 43, 21, 18, 17, 16, 15, 17. The same genotypes were measured in panels A-F, with genotype designations shown only in panels E and F. G,H,I) Hypocotyl epidermal cells of seedlings of indicated genotypes grown in short days for 2 days, visualized with the ML1:RFP shoot epidermis plasma membrane marker. Shown are z-stack confocal image projections of the near side of the hypocotyl. Scale bar, 0.1 mm. S2 Fig shows measurements of cell sizes in this experiment., Peer reviewed