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Plant Physiol, April 2000, Vol. 122, pp. 1003-1014

Activation Tagging in Arabidopsis1

Detlef Weigel,* Ji Hoon Ahn,2 Miguel A. Blázquez,2 Justin O. Borevitz,2 Sioux K. Christensen,2 Christian Fankhauser,2 Cristina Ferrándiz,2 Igor Kardailsky,23 Elizabeth J. Malancharuvil,2 Michael M. Neff,24 Jasmine Thuy Nguyen,25 Shusei Sato,2 Zhi-Yong Wang,2 Yiji Xia,2 Richard A. Dixon, Maria J. Harrison, Chris J. Lamb,6 Martin F. Yanofsky, and Joanne Chory

Plant Biology Laboratory (D.W., J.H.A., M.A.B., J.O.B., S.K.C., C.Fankhauser, I.K., E.J.M., M.M.N., J.T.N., Z.-Y.W., Y.X., C.J.L., J.C.) and Howard Hughes Medical Institute (J.C.), The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037; Department of Biology, University of California at San Diego, La Jolla, California 92093-0116 (C.Ferrándiz, S.S., M.F.Y.); and Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Parkway, Ardmore, Oklahoma 73402 (I.K., Y.X., R.A.D., M.J.H.)

Activation tagging using T-DNA vectors that contain multimerized transcriptional enhancers from the cauliflower mosaic virus (CaMV) 35S gene has been applied to Arabidopsis plants. New activation-tagging vectors that confer resistance to the antibiotic kanamycin or the herbicide glufosinate have been used to generate several tens of thousands of transformed plants. From these, over 30 dominant mutants with various phenotypes have been isolated. Analysis of a subset of mutants has shown that overexpressed genes are almost always found immediately adjacent to the inserted CaMV 35S enhancers, at distances ranging from 380 bp to 3.6 kb. In at least one case, the CaMV 35S enhancers led primarily to an enhancement of the endogenous expression pattern rather than to constitutive ectopic expression, suggesting that the CaMV 35S enhancers used here act differently than the complete CaMV 35S promoter. This has important implications for the spectrum of genes that will be discovered by this method.

1 This work was supported by grants from the National Science Foundation (no. MCB-9723823 to D.W., no. IBN-9728402 to M.F.Y., and no. MCB-9631390 to J.C.), the National Institutes of Health (no. R01 GM52413 to J.C.), and by the Samuel Roberts Noble Foundation. This work was also supported by a Research Experience for Undergraduates Supplement to National Science Foundation grant (no. IBN-9406948 to D.W.), and fellowships from the Korea Science and Engineering Foundation and the Hoffman Foundation (J.H.A.), the Spanish Ministry of Education (M.A.B., C.Ferrándiz), the National Science Foundation (S.K.C., Z.-Y.W.), the Human Frontiers Science Program Organization (M.A.B., C.Fa.), the Swiss National Science Foundation (C.Fankhauser), the National Institutes of Health (M.M.N.), and the Kazusa DNA Research Foundation (S.S.). D.W. was a National Science Foundation Young Investigator and J.C. was an Associate Investigator of the Howard Hughes Medical Institute.

2 These authors contributed equally to this study and are listed alphabetically.

3 Present address: Plant Gene Expression Center, 800 Buchanan Street, Albany, CA 94710.

4 Present address: Department of Biology, Washington University, One Brookings Drive, St. Louis, MO 63130.

5 Present address: Akkadix Corporation, 11099 North Torrey Pines Road, La Jolla, CA 92037.

6 Present address: John Innes Centre, Colney Lane, Norwich NR4 7UH, UK.

* Corresponding author; e-mail weigel{at}salk.edu; fax 858-558-6379.

© 2000 American Society of Plant Physiologists


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