Plant Cell Reports

, Volume 17, Issue 10, pp 752-759

First online:

Sonication-assisted Agrobacterium-mediated transformation of soybean immature cotyledons: optimization of transient expression

  • E. R. SantarémAffiliated withDepartment of Horticulture and Crop Science, Plant Molecular Biology and Biotechnology Program, OARDC, The Ohio State University, Wooster, OH, 44691, USA Fax: 330–263–3887 e-mail: finer.1@osu.edu
  • , H. N. TrickAffiliated withDepartment of Horticulture and Crop Science, Plant Molecular Biology and Biotechnology Program, OARDC, The Ohio State University, Wooster, OH, 44691, USA Fax: 330–263–3887 e-mail: finer.1@osu.edu
  • , J. S. EssigAffiliated withDepartment of Horticulture and Crop Science, Plant Molecular Biology and Biotechnology Program, OARDC, The Ohio State University, Wooster, OH, 44691, USA Fax: 330–263–3887 e-mail: finer.1@osu.edu
  • , J. J. FinerAffiliated withDepartment of Horticulture and Crop Science, Plant Molecular Biology and Biotechnology Program, OARDC, The Ohio State University, Wooster, OH, 44691, USA Fax: 330–263–3887 e-mail: finer.1@osu.edu

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Abstract

Sonication-assisted Agrobacterium-mediated transformation (SAAT) tremendously improves the efficiency of Agrobacterium infection by introducing large numbers of microwounds into the target plant tissue. Using immature cotyledons of soybean as explants, we evaluated the effects of the following parameters on transient β-glucuronidase (GUS) activity: cultivars, binary vectors, optical density of Agrobacterium during infection, duration of sonication treatment, co-culture conditions, length of explant preculture and addition of acetosyringone during co-culture. The extent of tissue disruption caused by sonication was also determined. The highest GUS expression was obtained when immature cotyledons were sonicated for 2 s in the presence of Agrobacterium (0.11 OD600nm) followed by co-cultivation with the abaxial side of the explant in contact with the culture medium for 3 days at 27°C. The addition of acetosyringone to the co-culture medium enhanced transient expression. No differences were observed when different cultivars or binary vectors were used. Cotyledons sonicated for 2 s had moderate tissue disruption, while the longer treatments resulted in more extensive damage.

Key wordsAgrobacterium tumefaciens Glycine max Transient expression SAAT Transformation