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Additional file 4. Primers used in this study.

Additional file 3. PCR amplification for transgenic-positive identification in calli. a Amplification of DsRed from calli of Jia 9709, Jia 2016, Zhong shuang 8, Zhong shuang 11 and Zhong you 821, respectively. P, plasmid. WT, wild type plant. Marker, DL 100 bp ladder. b Amplification of BnaA07g17400D from calli in 7633, B 351 and Shan 3B, respectively. P, plasmid. N, wild type plant. Marker, DL 2000 bp.

Additional file 1. Formation of calli to a regenerated plantlet on a hypocotyl. a and b calli that were induced from hypocotyls. c Calli that were prepared for swollen to shoots. d and e Shoot formation and cotyledon development accompanied by the appearance of a growth point.

Additional file 2. Visual screening of transgenic-positive and/or -negative tissues under a fluorescence stereomicroscope. Red fluorescence could be observed under fluorescence stereomicroscope to facilitate visual screening in tissue culture medium. The image was captured in bright field a and its corresponding dark field b, respectively. The tissues indicated by the arrow represent transgenic-positive calli.