Agrobacterium rhizogenes-mediated genetic transformation of radish (Raphanus sativus L. cv. Valentine) for accumulation of anthocyanin

作者：Bae Hanhong; Kim Yeon Bok; Park Nam Il; Kim Haeng Hoon; Kim Young Seon; Lee Mi Young; Park Sang Un^*

来源：Plant Omics, 2012, 5(4): 381-385.

• anthocyanin
• hairy root culture
• Raphanus sativus L. cv. Valentine
• genetic transformation
• 2,4-D

An efficient protocol for the transformation of radish (Raphanus sativus L. cv. Valentine) hairy root cultures was initiated using cotyledon explants and infected by Agrobacterium rhizogenes R1000, a strain with the binary vector pBI121. Kanamycin-resistant roots appeared on explants at 4 weeks after infection with A. rhizogenes and these roots were maintained on hormone-free selection medium. PCR analysis of the neomycin phosphotransferase (NTPII) gene confirmed transformation in 15 plants of kanamycin-resistant hairy root cultures from a total 20 plants. Detection of high levels of beta-glucuronidase (GUS) transcripts and enzyme activity, as well as GUS histochemical localization, also confirmed the stable genetic transformation. We inoculated isolated hairy roots in liquid medium to promote rapid growth and production of anthocyanin. Among the different auxin treatments, 1.0 mg/L 2,4-D treatment resulted the highest production of anthocyanin (30.9 mg/L) that was 26 times higher as compared to control (1.2 mg/L). Transgenic root cultures of R. sativus L. cv. Valentine will allow investigation of the molecular and metabolic regulation of anthocyanin biosynthesis and evaluation of the genetic engineering potential of this species.

• 出版日期2012-7