An early-flowering einkorn wheat mutant with deletions of PHYTOCLOCK 1/LUX ARRHYTHMO and VERNALIZATION 2 exhibits a high level of VERNALIZATION 1 expression induced by vernalization.

Using heavy-ion beam mutagenesis of Triticum monococcum strain KU104-1, we identified a mutant that shows extra early-flowering; it was named extra early-flowering 3 (exe3). Here, we carried out expression analyses of clock-related genes, clock downstream genes and photoperiod pathway genes, and found that the clock component gene PHYTOCLOCK 1/LUX ARRHYTHMO (PCL1/LUX) was not expressed in exe3 mutant plants. A PCR analysis of DNA markers indicated that the exe3 mutant had a deletion of wheat PCL1/LUX (WPCL1), and that the WPCL1 deletion was correlated with the mutant phenotype in the segregation line. We confirmed that the original strain KU104-1 carried a mutation that produced a null allele of a flowering repressor gene VERNALIZATION 2 (VRN2). As a result, the exe3 mutant has both WPCL1 and VRN2 loss-of-function mutations. Analysis of plant development in a growth chamber showed that vernalization treatment accelerated flowering time in the exe3 mutant under short day (SD) as well as long day (LD) conditions, and the early-flowering phenotype was correlated with the earlier up-regulation of VRN1. The deletion of WPCL1 affects the SD-specific expression patterns of some clock-related genes, clock downstream genes and photoperiod pathway genes, suggesting that the exe3 mutant causes a disordered SD response. The present study indicates that VRN1 expression is associated with the biological clock and the VRN1 up-regulation is not influenced by the presence or absence of VRN2.

Level of VERNALIZATION 1 expression is correlated with earliness in extra early-flowering mutant wheat lines.

Four extra early-flowering mutants, named extra early-flowering1 (exe1), exe2, exe3, and exe4, were identified in Triticum monococcum strain KU104-1 following heavy-ion beam mutagenesis. The four exe mutants fell into two groups, namely Type I (moderately extra early-flowering type; exe1 and exe3) and Type II (extremely extra early-flowering type; exe2 and exe4). Analysis of plant development in a growth chamber showed that the speed of leaf emergence was accelerated in exe mutants at the reproductive stage compared to wild-type (WT) plants. The speed of leaf emergence was faster in Type II than Type I plants. Analysis of VERNALIZATION 1 (VRN1), a flowering promoter gene, showed that it was more highly expressed in seedlings at early developmental stages in Type II mutants than Type I mutants. These findings indicate that the difference in earliness between Type I and Type II mutants is associated with the level of VRN1 expression. The original KU104-1 is an einkorn wheat strain that carries a null allele of the VRN2 gene, a repressor of flowering. Thus, our results indicate that the level of VRN1 expression controls earliness in exe mutants independently of VRN2.