Total RNA was isolated using Tiangen RNAprep plant kit (Tiangen, Beijing, China) from underground shoots collected from two-year-old bamboo plants and treated with RNase-free DNase I (Tiangen). cDNA was converted from 1 μg RNA using TransScript® II One-Step gDNA Removal and cDNA Synthesis SuperMix (TransGen Biotech).
Luciferase reporter assays were conducted as reported [62 (link)] with slight modifications, including vector, restriction enzyme and plasmid. PeGRF6 sequence with the predicted target site was inserted into the pGreenII 0800-miRNA vector [63 (link)] between KpnI and EcoRI. The precursor sequence of ped-miR396d-5p was inserted behind the 35S promoter of the pCAMBIA1300 vector. pGreenII-0800::PeGRF6 and 35S::ped-miR396d-5p vectors were transformed into Agrobacteria GV3101 (pSoup) and GV3101, respectively. The transformed strains were co-infiltrated into the leaves of N. benthamiana. Empty pCAMBIA1300 vectors were used as the controls. At 48 h after infiltration, fluorescent signals were detected and compared.Table S5 shows the primers.
Luciferase reporter assays were conducted as reported [62 (link)] with slight modifications, including vector, restriction enzyme and plasmid. PeGRF6 sequence with the predicted target site was inserted into the pGreenII 0800-miRNA vector [63 (link)] between KpnI and EcoRI. The precursor sequence of ped-miR396d-5p was inserted behind the 35S promoter of the pCAMBIA1300 vector. pGreenII-0800::PeGRF6 and 35S::ped-miR396d-5p vectors were transformed into Agrobacteria GV3101 (pSoup) and GV3101, respectively. The transformed strains were co-infiltrated into the leaves of N. benthamiana. Empty pCAMBIA1300 vectors were used as the controls. At 48 h after infiltration, fluorescent signals were detected and compared.
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