Extended Data Fig. 3: Role of NTL8 in vernalization and redundancy.
From: Temperature-dependent growth contributes to long-term cold sensing
a, Forward genetics identified ntl14-D. A schematic of NTL14 is shown below. The light blue box indicates the NAC domain, and the red box indicates TM. Q indicates glutamine, and the asterisk indicates the stop codon. Seven independent repeats with similar results were performed. b, Analysis of VIN3 transcript level by qPCR in the ntl8-1 mutant and the Col-0 control, at NV (20 °C) and 4 weeks of cold treatment (4W) at 5 °C. Error bars show s.e.m. of three biological replicates. c, As for b, but in ntl14-1 and the Ler control. Error bars show s.e.m. of four biological replicates. d, VIN3 transcript in ntl8-1ntl14-1 and WT control, at NV (20 °C) and 3 weeks of cold treatment (5 °C). WT, WT plants in the F2 population of the cross between ntl8-1 and ntl14-1. For comparison between WT and the ntl8-1ntl14-1 double mutant, we performed a two-tailed t-test: t = 4.634, d.f. =10, P = 0.0009. Error bars show s.e.m. of six biological replicates. e, f, Analysis of FLC transcripts in Ler JU223, vin3-6 JU223, ntl8-D2 JU223 and vin3-6 ntl8-D2 JU223 mutants under NV (e), 4WT0 (f) and 4WT14 (f) conditions. NV indicates the warm (20 °C) before cold treatment, 4W indicates 4 weeks of cold treatment at 5 °C, and 4WT14 indicates 4 weeks of cold plus 14 days of warm treatment at 20 °C. In f, data are normalized to the corresponding NV treatment. Error bars are s.e.m. of three biological replicates. g, Unspliced FLC levels after 4 weeks of vernalization in Col FRI and ntl8-1 FRI, normalized to the NV levels in the same genotype. Error bars are s.e.m. of three biological replicates. Unpaired t-test, two-tailed: t = 5.303, d.f. = 4, P = 0.0061. h, Flowering time, counted by the number of rosette leaves at flowering, for plants vernalized for 4 weeks. Error bars are s.e.m. of 12 individuals. Unpaired t-test, two-tailed: t = 4.241, d.f. = 22, P = 0.00031. i, Potential cross-regulation targets of NTL14 based on microarray data17. j, Potential cross-regulation targets of NTL8 based on in vitro data20. RNA was isolated from whole seedlings, and values are all normalized to UBC.
