Generations of knock-down mutants using RNA interfering technology
To generate RGB1 RNAi transgenic lines, a gene fragment of RGB1 was amplified from Nipponbare cDNA. A hairpin structure with two inverted repeat fragments was subsequently constructed and transferred into the plant binary vector pTCK303, expressed under the control of the maize ubiquitin promoter (Wang et al., 2004). The construct was transformed into the Agrobacterium strain EHA105 and used for plant transformation as described previously (Chen et al. , 2014). The DNA fragments embracing the targets of the plant transformants were amplified for sequencing.