A rapid and non-destructive screenable marker, FAST, for identifying transformed seeds of Arabidopsis thaliana
Article first published online: 25 NOV 2009
© 2009 The Authors. Journal compilation © 2009 Blackwell Publishing Ltd
The Plant Journal
Volume 61, Issue 3, pages 519–528, February 2010
How to Cite
Shimada, T. L., Shimada, T. and Hara-Nishimura, I. (2010), A rapid and non-destructive screenable marker, FAST, for identifying transformed seeds of Arabidopsis thaliana. The Plant Journal, 61: 519–528. doi: 10.1111/j.1365-313X.2009.04060.x
- Issue published online: 21 JAN 2010
- Article first published online: 25 NOV 2009
- Received 18 June 2009; revised 29 September 2009; accepted 13 October 2009; published online 25 November 2009.
- Arabidopsis thaliana;
- green fluorescent protein;
- screenable marker
The creation of transgenic plants has contributed extensively to the advancement of plant science. Establishing homozygous transgenic lines is time-consuming and laborious, and using antibiotics or herbicides to select transformed plants may adversely affect the growth of some transgenic plants. Here we describe a novel technology, which we have named FAST (fluorescence-accumulating seed technology), that overcomes these difficulties. Although this technology was designed for use in Arabidopsis thaliana, it may be adapted for use in other plants. The technology is based on the expression of a fluorescent co-dominant screenable marker FAST, under the control of a seed-specific promoter, on the oil body membrane. The FAST marker harbors a fusion gene encoding either GFP or RFP with an oil body membrane protein that is prominent in seeds. The marker protein was only expressed in a specific organ (i.e. in dry seeds) and at a specific time (i.e. during dormancy), which are desirable features of selectable and/or screenable markers. This technique provides an immediate and non-destructive method for identifying transformed dry seeds. It identified the heterozygous transformed seeds among the T1 population and the homozygous seeds among the T2 population with a false-discovery rate of <1%. The FAST marker reduces the length of time required to produce homozygous transgenic lines from 7.5 to 4 months. Furthermore, it does not require sterilization, clean-bench protocols or the handling of large numbers of plants. This technology should greatly facilitate the generation of transgenic Arabidopsis plants.