Agrobacterium-mediated large-scale transformation of wheat (Triticum aestivum L.) using glyphosate selection

被引：121

作者：

Hu, T ^{[1
]}

Metz, S ^{[1
]}

Chay, C ^{[1
]}

Zhou, HP ^{[1
]}

Biest, N ^{[1
]}

Chen, G ^{[1
]}

Cheng, M ^{[1
]}

Feng, X ^{[1
]}

Radionenko, M ^{[1
]}

Lu, F ^{[1
]}

Fry, J ^{[1
]}

机构：

[1] Monsanto Co, St Louis, MO 63017 USA

来源：

PLANT CELL REPORTS | 2003年 / 21卷 / 10期

关键词：

Agrobacterium transformation; wheat (Triticum aestivum L.); glyphosate selection; Roundup Ready;

D O I：

10.1007/s00299-003-0617-6

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

An Agrobacterium-mediated transformation system with glyphosate selection has been developed for the large-scale production of transgenic plants. The system uses 4-day precultured immature embryos as explants. A total of 30 vectors containing the 5-enol-pyruvylshikimate-3-phosphate synthase gene from Agrobacterium strain CP4 (aroA:CP4), which confers resistance to glyphosate, were introduced into wheat using this system. The aroA:CP4 gene served two roles in this study-selectable marker and gene of interest. More than 3,000 transgenic events were produced with an average transformation efficiency of 4.4%. The entire process from isolation of immature embryos to production of transgenic plantlets was 50-80 days. Transgenic events were evaluated over several generations based on genetic, agronomic and molecular criteria. Forty-six percent of the transgenic events fit a 3:1 segregation ratio. Molecular analysis confirmed that four of six lead transgenic events selected from Agrobacterium transformation contained a single insert and a single copy of the transgene. Stable expression of the aroA:CP4 gene was confirmed by ELISA through nine generations. A comparison of Agrobacterium-mediated transformation to a particle bombardment system demonstrated that the Agrobacterium system is reproducible, has a higher transformation efficiency with glyphosate selection and produces higher quality transgenic events in wheat. One of the lead events from this study, no. 33391, has been identified as a Roundup Ready wheat commercial candidate.

引用

页码：1010 / 1019

页数：10

共 48 条

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