African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12036

Full Length Research Paper

Transformation of multiple soybean cultivars by infecting cotyledonary-node with Agrobacterium tumefaciens

Xinping YI and Deyue YU*
Nanjing Agricultural University, National Center for Soybean Improvement, National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing 210095, China.
Email: [email protected]

  •  Accepted: 25 August 2006
  •  Published: 16 October 2006

Abstract

Transformation of four soybean cultivars (Nannong88-1, Nannong18-6, Yu23 and Nannong 87C-38) by infecting cotyledonary-node with Agrobacterium tumefaciensstrain EHA105 harboring pBI121 containing GFP reporter gene was conducted. The results indicated that the addition of thiol compounds (L-cysteine, dithiothreitol and sodium thiosulfate) in co-cultivation period increased the transformation efficiency of all four soybean cultivars, with Nannong 88-1 most increased up to 2.20%. Detection of GFP expression in the rooted plants was an effective selection system for the confirmation of soybean transformation. And most GFP-positive plants were confirmed to be positive by Southern blot analysis, which showed that transformation of cotyledonary-node explants mediated by Agrobacterium delivered T-DNA with one or two copies into soybean genome. In our study, the combination of Nannong88-1 with EHA105 is the optimum selection for explant and bacterial inoculum in soybean transformation, which could be applied in future functional study of soybean genes.

 

Key words: Soybean, cotyledonary–node, Agrobacterium, transformation, GFP.

Abbreviation

GFP, green fluorescent protein; BA, 6:benzylaminopurine; DTT,dithiolthreitol; IBA, indole-3-butyric acid