Plant transformation

To generate gene constructs with the C-terminal half of the catalytic subunit of DNA Pol ϵ, POLA5 (9), tagged with either GFP or HA epitopes, the corresponding cDNA fragments were cloned into pGWB6 or pEarleyGate201, respectively. The resulting clones were transformed in Col plants by Agrobacterium tumefaciens-mediated transformation using the floral-dip method (44). The Agrobacterium strain used was C58C1. The EMF2 cDNA was introduced in emf2-2/EMF2 heterozygous plants by floral-dip method and thus derived emf2-2 homozygous plants that expressed GFP-tagged EMF2 were selected on GM-glucose hygromycin-containing media plates. At least 10 independent transformants were evaluated for each construct. POLA5-HA lines were crossed with both 35S::GFP-CLFclf-50 and 35S::GFP-EMF2 emf2-2 plants.