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Abstract Detail

Plant biotech & Risk Assessment

Hsiao, Paoyuan [1], Sanjaya, Sanjaya [1], Su, Ruey-Chih [2], da Silva, Jaime A. Teixeira [3], Chan, Ming-Tsair [4].

Plant native tryptophan synthase beta 1 gene is a non-antibiotic selection marker for plant transformation.

Gene transformation is an integral tool for plant genetic engineering. All antibiotic resistant genes currently employed are of bacterial origin and their presence in the field is undesirable. Therefore, we developed a novel and efficient plant native non-antibiotic selection system for the selection of transgenic plants in the model system Arabidopsis. This new system is based on the enhanced expression of Arabidopsis tryptophan synthase beta 1 (AtTSB1) and the use of 5-methyl-tryptophan (5MT, a tryptophan [Trp] analog) and/or CdCl2 as selection agent(s). We successfully integrated an expression cassette containing an AtTSB1 cDNA driven by a cauliflower mosaic virus 35S promoter into Arabidopsis by floral dip transformation. Transgenic plants were efficiently selected on MS medium supplemented with 75 µM 5MT or 300 µM CdCl2 devoid of antibiotics. TSB1 selection was as efficient as the conventional hygromycin selection system. Northern blot analysis of transgenic plants selected by 5MT and CdCl2 revealed increased TSB1 mRNA transcript whereas uneven transcript levels of hygromycin phosphotransferase II (hpt) (control) was observed. Gas chromatography-mass spectrometry revealed 10-15 fold greater free Trp content in AtTSB1 transgenic plants than in wild-type plants grown with or without 5MT or CdCl2. Taken together, the TSB1 system provides a novel selection system distinct from conventional antibiotic selection systems.

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1 - Academia Sinica, Agricultural Biotechnology Research Center
2 - Fu Jen Catholic University, Department of Life Science
3 - Kagawa University, Department of Horticultural Science, Faculty of Agricultur
4 - Academia Sinica, Agricultural Biotechnology Research Center, 128, Academia Road, Sec. 2, Taipei, 11529, Taiwan

non-antibiotic marker
5' methyl-tryptophan (5MT)
Plant transformation
tryptophan synthase beta 1.

Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P45008
Abstract ID:738

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