Arabidopsis Mutants by Activation Tagging in which Photosynthesis Genes are Expressed in Dedifferentiated Calli
1 Laboratory of Plant Cell Technology and COE Program in the 21st Century, Graduate School of Nutritional and Environmental Sciences, University of Shizuoka, 52-1 Yada, Suruga, Shizuoka, 422-8526 Japan
2 National Institute for Basic Biology/Okazaki Institutes for Integrated Bioscience, Myodaiji-cho, Okazaki, 444-8585 Japan
3 Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Hongo, Tokyo, 113-0033 Japan
* Corresponding author: E-mail, hirokoba{at}u-shizuoka-ken.ac.jp; Fax, +81-54-264-5584.
In an effort to delineate the precise mechanisms underlying the organ-specific expression of photosynthesis genes, Arabidopsis lines homozygous for each transgene construct made with the gene for hygromycin B phosphotransferase or ß-glucuronidase (GUS) placed under control of the promoter of the nuclear gene for the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (RBCS-3B) were constructed. Furthermore, activation tagging with T-DNA possessing quadruply repeated enhancers derived from the cauliflower mosaic virus 35S promoter was applied to a transgenic line of Arabidopsis. Mutants resistant to hygromycin B during the growth of calli generated from non-green roots on callus-inducing medium resulted from the expression of hygromycin B phosphotransferase driven by the RBCS-3B promoter. Three mutant lines, ces101 to ces103 (callus expression of RBCS), were obtained from approximately 4,000 calli resistant to a selectable marker for transformation. The active transcription driven by the RBCS-3B promoter in all the calli of ces mutants was confirmed by expression of both the GUS reporter gene and endogenous RBCS-3B. Chlorophyll and carotenoids, as well as light-dependent O2 evolution, have been detected in the calli of all ces mutants. The loci where T-DNA was integrated in the ces101 line were determined by thermal asymmetric interlaced (TAIL)-PCR. The introduction of a DNA fragment harboring the gene for receptor-like kinase placed under the influence of enhancers into the parental line reproduced the phenotype of ces mutants. We have thus concluded that CES101 is a receptor-like kinase. The strategy presented in this investigation may promise to select a greater number of ces mutants.
4 These authors contributed equally to this work.
5 Present address: Kumiai Chemical Industry Co. Ltd, Life Science Research Institute, 276 Tamari, Kakegawa, Shizuoka, 436-0011 Japan.
6 Present address: Lotte Central Laboratory Co., Ltd, 3-1-1 Numakage, Saitama, Saitama, 336-8601 Japan.
7 Present address: Department of Biology, University of Utah, 257 South 1400 East, Salt Lake City, UT 84112-0840, USA.
8 Adjunct addresses: Graduate University for Advanced Studies, Shonan Village, Hayama, Kanagawa, 240-0193 Japan; and Graduate School of Science, Kyoto University, Kyoto, 606-8502 Japan.
(Received September 5, 2005; Accepted December 7, 2005)
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  E. Lopez-Juez Plastid biogenesis, between light and shadows J. Exp. Bot., January 1, 2007; 58(1): 11 - 26. [Abstract] [Full Text] [PDF]
|
|