Allotetraploids were resynthesized as previously described1 (link),4 (link), and hybrids were made by crossing C24 with Columbia. Unless noted otherwise, 8−15 plants (grown under 22°C and 16-hour light/day) from each of 2−3 biological replications were pooled for the analysis of DNA, RNA, protein, chlorophyll, starch, and sugar. TOC1:CCA1 and TOC1:cca1-RNAi transgenic plants were produced using pEarlygate303 (CD694) and pCAMBIA (CD3−447) derivatives, respectively. cca1−11 (CS9378) and ccal-11 lhy-21 (CS9380) mutants9 (link),22 (link),29 (link) were obtained from ABRC. Protein blot, EMSA, and ChIP assays were performed as previously described17 (link),18 (link).
Full Methods and any associated references are available in the online version of the paper at www.nature.com/nature .
Protocol detail
Allotetraploid Resynthesis and Hybrid Production
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Biological
Chip assays
Chlorophyll
Derivatives
Emsa
Hybrids
Light
Plants
Protein
Replications
Rnai
Starch
Sugar
Transgenic plants
Corresponding Organization :
Other organizations : China Agricultural University, Institute of Molecular and Cell Biology
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Variable analysis
independent variables
- Genotype (allotetraploids, hybrids, TOC1:CCA1 transgenic plants, TOC1:cca1-RNAi transgenic plants, cca1-11 mutants, cca1-11 lhy-21 mutants)
- Protein
- Chlorophyll
- Starch
- Sugar
- Temperature (22°C)
- Photoperiod (16-hour light/day)
- Replication (2-3 biological replicates)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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