Trehalose-6-phosphate synthase as an intrinsic selection marker for plant transformation

被引:21
|
作者
Leyman, B
Avonce, N
Ramon, M
Van Dijck, P
Iturriaga, G
Thevelein, JM
机构
[1] Katholieke Univ Leuven VIB, Dept Mol Microbiol, B-3001 Heverlee, Belgium
[2] Katholieke Univ Leuven, Inst Bot & Microbiol, Mol Cell Biol Lab, B-3001 Heverlee, Belgium
[3] UAEM, Ctr Invest Biotecnol, Cuernavaca 62210, Morelos, Mexico
关键词
plant transformation; intrinsic selection marker; AtTPS1; trehalose-6-phosphate synthase; glucose resistance; Arabidopsis thaliana; Nicotiana tabacum;
D O I
10.1016/j.jbiotec.2005.08.033
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Insertion of foreign DNA into plant genomes occurs randomly and with low frequency. Hence, a selectable marker is generally required to identify transgenic plants. Until now, all selection systems have been based on the use of non-plant genes, derived from microorganisms and usually conferring antibiotic or herbicide resistance. The use of microorganism-derived genes however has raised biosafety concerns. We have developed a novel selection system based on enhancing the expression of a plant-intrinsic gene and the use of a harmless selection agent. Selection takes advantage of the reduced glucose sensitivity of seedlings with enhanced expression of AtTPS1, a gene encoding trehalose-6-P synthase. As a result, transformants can be identified as developing green seedlings amongst the background of small, pale non-transformed plantlets on high glucose medium. In addition, vegetative regeneration of tobacco leaf explants is very sensitive to high external glucose. Overexpression of AtTPS1 in tobacco allows selecting glucose insensitive transgenic shoots. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:309 / 317
页数:9
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