Research data supporting "GIGANTEA is required for robust circadian rhythms in wheat"

The data describe studies investigating the role of GIGANTEA (GI) in wheat. We isolated TILLING mutants in the tetraploid wheat variety T. turgidum cv. Kronos. Two TILLING lines with premature stop codons within the A GI homoeologues (GI-A, TraesCS3A02G116300, Kronos2019) and B GI homoeologues (GI-B, TraesCS3B02G135400, Kronos2205) genes were obtained from the Germplasm Resource Unit (GRU), Norwich UK (Figure 1). Kronos2019 contains a C to T SNP 6606 nucleotide base pairs (bp) from the start of the GI-A transcription start site (TSS) (position 3A:84190982 IWGSC Refseq V1.1). This SNP is situated within the 12th exon of the first splice variant and the fourth exon of the second splice variant and is predicted to result in the deletion of 356 amino acids (Fig. 1A). Kronos2205 contains a G to A SNP 6348 bp downstream of the GI-B TSS (position 3B:117929486 IWGSC Refseq V1.1). This SNP is situated within the 12th exon of the first two splice variants and the 11th exon of the third splice variant and is predicted to result in the deletion of 403 amino acids (Fig. 1B). It is likely that both mutations would lead to nonsense mediated mRNA decay as the nearest intron to these SNPs is over 50 bp away (Nyikó et al., 2013). Kronos TILLING lines were backcrossed four times to Kronos. The BC4 F1 heterozygous plants were self-crossed to produce BC4 F2 seed. BC4 F2 seeds were genotyped, and seeds retained from plants which were homozygous for the Kronos2019 or Kronos2205 mutations. The homozygous Kronos2019 (Ttgi-A1) and Kronos2205 (Ttgi-B1) lines were crossed, self-crossed and plants homozygous for both Kronos2019 and Kronos2205 mutations (Ttgi-A1/gi-B1) were selected. To control for possible background mutations, that are present in the TILLING populations, plants homozygous for both wild type alleles were selected as mutant sibling wild type (WT segregant). Kronos wild type line was selected as non-mutagenized wild type control. The data are in Excel spreadsheet, with each sheet describing a separate experiment. Brief descriptions are on the description page. Fuller details below. Sheet 2 Non photochemical quenching of Kronos, WT segregant, Ttgi-B1, Ttgi-A1 and TtgiA1/gi-B1 leaf fragments in constant light (n = > 10). Sheet 3 Mean abundance of circadian oscillator transcripts (n = 3-5) in Ttgi-A1/gi-B1 and Kronos WT ). Transcript abundance (ΔΔCq) is relative to TtRP15 and TtRPT5A.. The data are described for sample points in hours. Sheet 4 Flowering time and yield data for Ttgi-A1/gi-B1, Ttgi-A1, Ttgi-B1, WT segregant and Kronos grown in long photoperiod (LD, 16 h light at 250 µmol m−2 s−1 , 20°C: 8 h dark 16°C) and short photoperiod (LD, 8 h light at 250 µmol m−2 s−1, 20°C: 16 h dark 16°C). GS55 is used as reference for heading date and defined as days after sowing (DAS). Sheet 5 Flowering time in the field and yield data for. Kronos, WT segregant, Ttgi-A1 and Ttgi-A1/gi-B1 grown in the field at NIAB experimental farm in the 2021 field season. Heading date as defined as days after sowing to reach GS55. Sheet 6 Transcript abundance of wheat genes involved in flowering in Ttgi-A1/gi-B1 and Kronos WT grown under long day (16 h light at 250 µmol m−2 s−1, 20°C: 8 h dark 16°C). Once plants reached the 3-leaf stage or GS39, sampling of the first true leaf commenced at time 0 to 24 hours, every 3 hours. Sheet 7 The chlorophyll a parameter, Fv/Fm in Ttgi-A1/gi-B1 in constant conditions (light and temperature). Kronos, WT Segregant, Ttgi-B1, Ttgi-A1 and TtgiA1/gi-B1 (n= > 10). Sheet 8 Kronos, WT segregant, Ttgi-A1, Ttgi-B1 and Ttgi-A1/gi-B1 plants were grown under long photoperiod (LD, 16 h light at 250 µmol m−2 s−1, 20°C: 8 h dark 16°C). The following were measured Plant length.) Number of tillers. Length of the primary head. Number of seeds per primary head. Weight of seeds produced by the primary head. Total number of seeds produced by each plant. Sheet 9 Kronos, WT segregant, Ttgi-A1, Ttgi-B1 and Ttgi-A1/gi-B1 plants were grown under short photoperiod (SD, 8 h light at 250 µmol m−2 s−1, 20°C: 16 h dark 16°C). The following were measured Plant length.) Number of tillers. Length of the primary head. Number of seeds per primary head. Weight of seeds produced by the primary head. Total number of seeds produced by each plant. Sheet 10 Morphological and yield traits in TtGI lines and Kronos in field conditions at NIAB farm. Kronos, WT segregant, Ttgi-A1, Ttgi-B1 and Ttgi-A1/gi-B1 plants were grown in NIAB experimental farm during the 2020 field season. Height. Length of primary head. Number of seeds of representative head. Mean weight of seeds of representative head per plot. Sheet 11 Abundance of transcripts expression of genes involved in gibberellin synthesis in Ttgi-A1/gi-B1 and Kronos WT grown under long day (16 h light at 250 µmol m−2 s−1, 20°C: 8 h dark 16°C). Once plants reached 3-leaves or GS39, sampling of the first true leaf commenced at time 0 to 24 hours every 3 hours. Transcript abundance (ΔΔCq) is relative to TtRP15 and TtRPT5A,