Fig. 2: HtL1 accelerates flowering in winter wheat.

a The morphological phenotypes of HtL1-OE, HtL1-RNAi and wild-type KN199 plants. Seeds were initially germinated and cultivated in the dark at room temperature for 2 to 3 days. Subsequently, the materials were vernalized at 4 °C for 21 days, after which they were transplanted into the greenhouse. Flowering phenotypes of HtL1-OE and HtL1-RNAi lines were observed and photographed at 73 and 80 days, respectively. Scale bars, 10 cm. b Statistical analysis of heading time of HtL1-OE, HtL1-RNAi and KN199 plants. The box plots display the interquartile range, comprising the first quartile, median, and third quartile, while the whiskers extend from the minimum to the maximum values. Two-tailed Student’s t-test for statistical analysis, (n = 20 plants for each line). c and d The relative mRNA abundance of HtL1 and VRN1 in HtL1-OE and HtL1-RNAi lines, as well as KN199. RNA were extracted from plumules of wheat plants vernalized for 21 days, and transcription levels of the indicated genes were analyzed by RT-qPCR. The data were normalized to ACTIN, then normalized to KN199. Independent biological experiments were conducted three times. Data are mean ± SD, two-tailed Student’s t-test for statistical analysis. e and f The monomeric and homotetrameric structures of HtL1 predicted by SWISS-MODEL. g and h Enzyme kinetic curve and activity assays of in vitro purified HtL1. Three independent biological replicates were performed. Data are presented as mean ± SD, two-tailed Student’s t-test for statistical analysis. i Aldolase activity analysis in HtL1-OE and HtL1-RNAi plants with 21 days of vernalization treatment. Three independent biological replicates were performed. Data are mean ± SD, two-tailed Student’s t-test for statistical analysis.