Screening of the rice FOX Arabidopsis lines for various phenotypes led to the identification of several lines exhibiting the desired phenotypes. To determine whether the rice cDNA inserts in these transgenic lines could generate the same phenotypes in rice, we introduced these cDNAs under the control of a maize ubiquitin promoter into rice for subsequent verification of the target phenotypes.
Full-length rice cDNAs were obtained from the Rice Genome Resource Center of the National Institute of Agrobiological Resources or from PCR products of genomic DNA isolated from selected rice FOX Arabidopsis hunting lines as templates. The DNA sequences were cut with SfiI and inserted into the SfiI site of pBIGS-RiceFOX. The resultant plasmids were introduced into Agrobacterium tumefaciens EHA105 by electroporation.
Nipponbare (Oryza sativa ssp. japonica) was transformed with a high-speed transformation system (Toki et al., 2006), and plants resistant to hygromycin (50 µg/mL) were selected. Regenerated plants were grown in isolated greenhouses at 28 °C. Aboveground visible phenotypes were observed in the T0 generation. Detailed phenotypes were observed in the T1 generation.