A previously described construct with the Col-0 genomic FLM fragment pFLM::gFLM template (pDP34) (Posé et al., 2013) was recombined into a pDONR201 destination vector using the Gateway system (Life Technologies, Carlsbad, CA). This vector was used as a template (FLMCol-0) to generate mutations using either a single phosphorylated primer or a combination of forward and reverse primers (Sawano and Miyawaki, 2000; Hansson et al., 2008). In case of variants with multiple modifications, individual mutations were introduced one at a time. The mutated inserts were recombined to the pFAST-R07 expression vector using the Gateway system (Life Technologies, Carlsbad, CA) (Shimada et al., 2010). All expression constructs were verified by sequencing and transformed into Agrobacterium tumefaciens strain GV3101. Nd-1 plants were transformed using the floral-dip method (Clough and Bent, 1998) and transgenic plants were identified based on seed fluorescence (Shimada et al., 2010). Segregation of T2 lines was examined and lines with single insertion events were selected for further analysis based on segregation ratios (Shimada et al., 2010). A list of primers and expression constructs is listed in Supplementary file 7.