new time-saving transformation system for Brassica napus

  • F Kong
  • J Li
  • X Tan
  • L Zhang
  • Z Zhang
  • C Qi
  • X Ma
Keywords: Agrobacterium tumefactiens, cotyledonary node, 6-benzyladenine, naphthalene acetic acid, transformation, Silwet L 77.

Abstract

By optimizing different parameters for infection, an efficient Agrobacterium-mediated transformation system for Brassica napus was developed. A series of combinations of optical densities, infection durations, concentrations of acetosyringone and silwet L77 were evaluated. Maximum transformation efficiency reached 18.93% when cotyledonary nodes were infected for 10 min with an Agrobacterium suspension of 0.8 OD and 200 ìM acetosyingone and 0.02% silwet L77 in the culture medium. In addition, the concentrations of 6-benzyladenine and naphthalene acetic acid were also optimized. Meanwhile, maximum transformation efficiency for green plantlet was 94.38% when cotyledonary nodes were cultivated in the medium containing 4 mg/L 6-benzyladenine and 0.04 mg/L naphthalene acetic after co-cultivation. Transformation was confirmed by polymerase chain reaction analysis, southern blotting analysis and green fluorescent protein assay.
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eISSN: 1684-5315