Summary
A procedure for the regeneration of fertile transgenic white mustard (Sinapis alba L.) is presented. The protocol is based on infection of stem explants of 7–9 day old plants with an Agrobacterium tumefaciens strain harboring a disarmed binary vector with chimeric genes encoding neomycin phosphotransferase and β-glucuronidase. Shoots are regenerated from callus-forming explants within 3–4 weeks. Under selection, 10% of the explants with transgenic embryonic callus develop into fertile transgenic plants. Rooting shoots transferred to soil yield seeds within 14–16 weeks following transformation. Integration and expression of the T-DNA encoded marker genes was confirmed by histochemical β glucuronidase assays and Southern-DNA hybridization using primary transformants and S1-progeny. The analysis showed stable integration and Mendelian inheritance of trans-genes in transformed Sinapis lines.
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Abbreviations
- BAP:
-
6-benzylaminopurine
- CaMV:
-
cauliflower mosaic virus
- GUS:
-
β-glucuronidase
- IBA:
-
indole-3-butyric acid
- IM:
-
infection medium
- NAA:
-
1-naphthalene acetic acid
- neo :
-
gene encoding NPTII
- NPTII:
-
neomycin phosphotransferase
- RIM:
-
root-inducing medium
- SEM:
-
shoot-elongation medium
- SIM:
-
shoot-inducing medium
- t-nos:
-
polyadenylation site of the nopaline synthase gene
- uidA :
-
gene encoding GUS
- WM:
-
wash medium
- X-Gluc:
-
5-bromo-4-chloro-3-indolyl β-D-glucuronide
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Hadfi, K., Batschauer, A. Agrobacterium-mediated transformation of white mustard (Sinapis alba L.) and regeneration of transgenic plants. Plant Cell Reports 13, 130–134 (1994). https://doi.org/10.1007/BF00239878
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DOI: https://doi.org/10.1007/BF00239878