Establishment of Brassica rapa rapa hairy root lines and
their maintenance
The eGFP Brassica rapa rapa hairy root lines were developed by
BIOPI (Plant Biology and Innovation, Amiens, France) resulting from
transformation with a construct portraying a double 35S promoter-SP-His
tag-eGFP-CaMV polyA as described by Huet et al (2014). Similarly, the
human alpha-L-iduronidase (IDUA) B. rapa rapa hairy root line was
developed with the same vector set up but as a gene of interest encoding
IDUA, as presented by Cardon et al. (2019).
Fresh hairy roots were subcultured every three weeks onto solid modified
Gamborg B5 medium containing 10-fold higher concentrations of copper
sulfate and cobalt chloride, and no casein when compared to the original
Gamborg B5 (Oksman-Caldentey et al., 1991) and grown in the dark at
+24°C. Prior to inoculation of liquid media, the hairy roots were
subcultured on solid plates for seven days as a standardized time.