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

• Published in: Plant cell reports 2006-12, Vol.25 (12), p.1355-1361
• Main Authors: Nishizawa, Keito, Kita, Yoichi, Kitayama, Masahiko, Ishimoto, Masao
• Format: Article
• Language: English
• Subjects: Aequorea victoria; Animals; Biological and medical sciences; Biotechnology; Blotting, Southern; Desiccation; Discosoma; DNA, Plant - metabolism; Electrophoresis, Polyacrylamide Gel; Fluorescence; Fluorescence microscopy; Fundamental and applied biological sciences. Psychology; Gene Expression; Genes, Reporter; Genetic engineering; Genetic technics; Genetic Vectors; Genome, Plant - genetics; Glycine max; Glycine max - genetics; Luminescent Proteins - analysis; Luminescent Proteins - metabolism; Methods. Procedures. Technologies; Plants, Genetically Modified; Plasmids; Proteins; Red Fluorescent Protein; Regeneration; Seeds; Seeds - metabolism; Soybeans; Transformation, Genetic; Transgenic animals and transgenic plants; Transgenic plants
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-006-0210-x

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.