Reef-coral proteins as visual, non-destructive reporters for plant transformation

• Published in: Plant cell reports Vol. 22; no. 4; pp. 244 - 251
• Main Authors: WENCK, A, PUGIEUX, C, WANG, W. C, REED, J, DRAYTON, P, OLIVER, D, TRAFFORD, H, LEGRIS, G, RUSHTON, H, TAYAB, S, LAUNIS, K, CHANG, Y.-F, TURNER, M, CHEN, D.-F, MELCHERS, L, DUNN, M, STACY, C, TIOZZO, A, DUNDER, E, VAN GRINSVEN, E, KHAN, R, SIGAREVA, M
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01.11.2003; Springer Nature B.V
• Subjects: Agronomy. Soil science and plant productions; Anthozoa; Biological and medical sciences; Biotechnology; Corn; Fluorescence; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic engineering applications; Genetic Markers; Genetic technics; Genetics and breeding of economic plants; Luminescent Proteins - genetics; Methods. Procedures. Technologies; Plant breeding: fundamental aspects and methodology; Plants - genetics; Proteins; Soybeans; Tomatoes; Transformation, Genetic; Transgenic animals and transgenic plants; Transgenic plants; Wavelengths; Fluorescent proteins· Plant transformation· Visual markers; Genetic transformation; Genetic marker; AmCyan; Interspecific comparison; Fluorescence; Coral reef; Gene expression; Protein; ZsGreen· ZsYellow; DsRed; Reporter gene; AsRed; Transgenic plant
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s00299-003-0690-x

Recently, five novel fluorescent proteins have been isolated from non-bioluminescent species of reef-coral organisms and have been made available through ClonTech. They are AmCyan, AsRed, DsRed, ZsGreen and ZsYellow. These proteins are valuable as reporters for transformation because they do not require a substrate or external co-factor to emit fluorescence and can be tested in vivo without destruction of the tissue under study. We have evaluated them in a large range of plants, both monocots and dicots, and our results indicate that they are valuable reporting tools for transformation in a wide variety of crops. We report here their successful expression in wheat, maize, barley, rice, banana, onion, soybean, cotton, tobacco, potato and tomato. Transient expression could be observed as early as 24 h after DNA delivery in some cases, allowing for very clear visualization of individually transformed cells. Stable transgenic events were generated, using mannose, kanamycin or hygromycin selection. Transgenic plants were phenotypically normal, showing a wide range of fluorescence levels, and were fertile. Expression of AmCyan, ZsGreen and AsRed was visible in maize T1 seeds, allowing visual segregation to more than 99% accuracy. The excitation and emission wavelengths of some of these proteins are significantly different; the difference is enough for the simultaneous visualization of cells transformed with more than one of the fluorescent proteins. These proteins will become useful tools for transformation optimization and other studies. The wide variety of plants successfully tested demonstrates that these proteins will potentially find broad use in plant biology.