Two critical factors are required for efficient transformation of multiple soybean cultivars: Agrobacterium strain and orientation of immature cotyledonary explant

Tae Seok Ko, Sangman Lee, Sergei Krasnyanski, Schuyler S. Korban

Research output: Contribution to journalArticlepeer-review

Abstract

An efficient transformation system was developed for multiple soybean [Glycine max (L.) Merrill.] cultivars using Agrobacterium-mediated gene transfer. A significantly high number of hygromycin-resistant somatic embryos (SEs) was obtained when immature zygotic cotyledons were inoculated with Agrobacterium tumefaciens strain KYRT1 and when the abaxial side of explants was oriented upwards (i.e., the adaxial side of explants was in contact with the medium). Most hygromycin-resistant SEs on selective medium were induced along the periphery of the abaxial side of cotyledonary explants. Extended periods of selection (up to 10 weeks post-cocultivation) increased the frequency of somatic embryogenesis, and more than 50% of selected SEs tested positive for β-glucuronidase (GUS). Following maturation and regeneration of selected SEs, ten independent transgenic soybean plants of cv Jack were obtained, and the overall transformation frequency ranged from 1.1 to 1.7%. Six and two transgenic plantlets were obtained from cvs Dwight and Williams, respectively. In addition, transgenic suspension lines were established from cvs Jack, Williams, Dwight, Rend and Ina. Molecular analysis of embryogenic lines and/or transgenic plants, established from different cultivars, confirmed stable integration, expression, and/or inheritance of transgenes in both T 0 and T1 plants.

Original languageEnglish (US)
Pages (from-to)439-447
Number of pages9
JournalTheoretical and Applied Genetics
Volume107
Issue number3
DOIs
StatePublished - Aug 1 2003

Keywords

  • Agrobacterium tumefaciens
  • Genetic transformation
  • Glycine max
  • Immature zygotic cotyledons
  • KYRT1
  • Soybean

ASJC Scopus subject areas

  • Biotechnology
  • Agronomy and Crop Science
  • Genetics

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