Supplement to A Distorted Circadian Clock Causes Early Flowering and Temperature-Dependent Variation in Spike Development in the Eps-3Am Mutant of Einkorn Wheat | Genetics
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Research ArticleINVESTIGATION

A Distorted Circadian Clock Causes Early Flowering and Temperature-Dependent Variation in Spike Development in the Eps-3Am Mutant of Einkorn Wheat

Piotr Gawroński, Ruvini Ariyadasa, Axel Himmelbach, Naser Poursarebani, Benjamin Kilian, Nils Stein, Burkhard Steuernagel, Goetz Hensel, Jochen Kumlehn, Sunish Kumar Sehgal, Bikram S. Gill, Peter Gould, Anthony Hall and Thorsten Schnurbusch
Genetics January 2014, genetics.113.158444; DOI: https://doi.org/10.1534/genetics.113.158444

Files in this data supplement:

  • Supporting Information: Figures S1-S4, Tables S1-S10, and Files S1-S13 (PDF, 683 KB)
  • Figure S1: TILLING of HvPUMILIO in barley cultivar 'Barke' revealed an M4 family (11266) with a knock-out mutation. (PDF, 369 KB)
  • Figure S2: Results from DNA gel blot hybridization showing deletion of the two genes, TmPUMILIO and TmLUX from the genome of the KT3-5 mutant in einkorn wheat. (PDF, 468 KB)
  • Figure S3: Relative transcript levels of circadian clock genes in transgenic barley. (PDF, 396 KB)
  • Figure S4: Allele mining in a diverse wheat collection revealed a putative mutant at TtLUX-A in cultivar 'Tsing Hua no. 559'. (PDF, 440 KB)
  • Table S1: List of primers. (PDF, 134 KB)
  • Table S2: Gene content in the physical contig_95 from barley and contigs 1331 and 1512 from bread wheat syntenic to the Eps-3Am locus. (PDF, 166 KB)
  • Table S3: KT lines of einkorn wheat analyzed. (PDF, 126 KB)
  • Table S4: Haplotype marker analysis performed on 15 KT mutants at the Eps-3Am locus. (PDF, 159 KB)
  • Table S5: Selected results from the delayed fluorescence (DF) measurements performed on T. monococcum wild type KT3-1 and mutant KT3-5. (PDF, 155 KB)
  • Table S6: Phenotypic data of transgenic HvLUX knock-down (RNAi) plants as compared to cv. 'Golden Promise'. (PDF, 141 KB)
  • Table S7: Results from re-sequencing of the wheat LUX in the collection of 96 accessions. (PDF, 140 KB)
  • Table S8: Selected wheat accessions grouped according to the cultivation status (wild or domesticated) to assess variation in the LUX sequence, expressed as a number of haplotypes. (PDF, 123 KB)
  • Table S9: Selected wheat accessions grouped according to the climatic condition at the site of origin (cool or warm) to assess variation in the LUX sequence, expressed as a number of haplotypes. (PDF, 123 KB)
  • Table S10: Selected wheat accessions grouped according to the climatic condition at the site of origin (cool or warm) and cultivation status (wild or domesticated) to assess variation in the LUX sequence, expressed as a number of haplotypes. (PDF, 122 KB)
  • File S1: Materials and Methods (PDF, 187 KB)
  • File S2: Phenotypic characterization of T. monococcum wild type line KT3-1 and mutant line KT3-5. Plants were grown in a controlled chamber under three different regimes: short day (SD), long day (LD) cool and LD warm. Data were collected from at least seven plants/genotype/regime. (.xls, 33 KB)
  • File S3: Mapping of the Eps-3Am locus in F2 and F3 populations. Molecular markers INDEL_201_202 and INDEL_271_272 flanking the Eps-3Am locus were used to screen F2 population and select 38 F2 recombinants. Plants were further genotyped with additional PCR-based markers to narrow down the locus. Seeds of eight F2 individuals with closest recombination events were sown to analyze their F3 progeny. Days to heading were scored for each F2 and F3 plant. (.xls, 80 KB)
  • File S4: Assembled BAC 454 sequence reads from barley contig_95. Eps-3Am locus from einkorn wheat was linked to the syntenic part of barley chromosome 3H. Selected contig_95 from a physical map of barley was sequenced following the 454 technology. (.txt, 654 KB)
  • File S5: DNA and protein sequences of putative HvPUMILIO mutant in barley cv. 'Barke'. (PDF, 145 KB)
  • File S6: Assembled BAC 454 sequence reads from bread wheat contig_1331. Eps-3Am locus from einkorn wheat was linked to the syntenic part of bread wheat chromosome 3AL. Selected contig_1331 from a physical map of bread wheat was sequenced following the 454 technology. (.txt, 1 MB)
  • File S7: Assembled BAC 454 sequence reads from bread wheat contig_1512. Eps-3Am locus from einkorn wheat was linked to the syntenic part of bread wheat chromosome 3AL. Selected contig_1512 from a physical map of bread wheat was sequenced following the 454 technology. (.txt, 301 KB)
  • File S8: Numerical light intensities from the delayed fluorescence (DF) measurements in einkorn wheat. Wild type line KT3-1 and mutant KT3-5 were imaged using an ORCA-II-BT 1024 16-bit camera (Hamamatsu Photonics, Japan) cooled to -80 degrees. The pictures were analyzed in the Metamorph 6.0 (Universal Imaging Corp., Downingtown, USA) and the numerical light intensities were extracted. (.xls, 177 KB)
  • File S9: Transcript levels of circadian clock genes in einkorn wheat, transgenic barley plants and durum wheat. qPCR reactions were performed on the ABI 7900HT Fast Real-Time PCR System (Applied Biosystems) employing QuantiTect SYBR Green PCR KIT (Qiagen) (.xls, 1 MB)
  • File S10: Phenotypic characterization of HvLUX-RNAi plants in barley cv. 'Golden Promise'. Putative knock down plants (BG284E10, BG284E11, BG353_1E15, BG353_1E01, BG353_2E06) and DH lines (DH_BG284E11_PP1, DH_BG284E11_PP7), and cv. 'Golden Promise' were grown under two different regimes. Days to heading were scored for every single plant. (.xls, 38 KB)
  • File S11: Panel of 96 diverse wheat accessions characterized by heading date and the haplotype of LUX. "Climate" was estimated based on either collection site or the country of origin. (.xls, 63 KB)
  • File S12: DNA sequences of 21 LUX-A haplotypes found among 96 wheat accessions (.txt, 14 KB)
  • File S13: DNA sequences of 8 LUX-B haplotypes found among 96 wheat accessions (.txt, 6 KB)

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