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Plant Cell Reports

, Volume 25, Issue 12, pp 1355–1361 | Cite as

A red fluorescent protein, DsRed2, as a visual reporter for transient expression and stable transformation in soybean

  • Keito Nishizawa
  • Yoichi Kita
  • Masahiko Kitayama
  • Masao Ishimoto
Genetic Transformation and Hybridization

Abstract

Fluorescent proteins such as green fluorescent protein (GFP) from Aequorea victoria are often used as markers for transient expression and stable transformation in plants, given that their detection does not require a substrate and they can be monitored in a nondestructive manner. We have now evaluated the red fluorescent protein DsRed2 (a mutant form of DsRed from Discosoma sp.) for its suitability as a visual marker in combination with antibiotic selection for genetic transformation of soybean [Glycine max (L.) Merrill]. Transient and stable expression of DsRed2 in somatic embryos was readily detected by fluorescence microscopy, allowing easy confirmation of gene introduction. We obtained several fertile transgenic lines, including homozygous lines, that grew and produced seeds in an apparently normal manner. The red fluorescence of DsRed2 was detected by fluorescence microscopy without background fluorescence in both leaves and seeds of the transgenic plants. Furthermore, in contrast to seeds expressing GFP, those expressing DsRed2 were readily identifiable even under white light by the color conferred by the transgene product. The protein composition of seeds was not affected by the introduction of DsRed2, with the exception of the accumulation of DsRed2 itself, which was detectable as an additional band on electrophoresis. These results indicate that DsRed2 is a suitable reporter (even more suitable than GFP) for genetic transformation of soybean.

Keywords

Bombardment DsRed2 Fluorescent protein Nondestructive Transgenic soybean 

Notes

Acknowledgments

This work was supported by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN). We thank Y. Niwa for providing the sGFP(S65T) reporter gene.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Keito Nishizawa
    • 1
  • Yoichi Kita
    • 2
  • Masahiko Kitayama
    • 2
  • Masao Ishimoto
    • 1
  1. 1.National Agricultural Research Center for Hokkaido RegionHokkaidoJapan
  2. 2.Ehime Women’s CollegeEhimeJapan

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