The ability of pea transformation technology to transfer genes into peas adapted to western Canadian growing conditions

被引:40
作者
Polowick, PL [1 ]
Quandt, J [1 ]
Mahon, JD [1 ]
机构
[1] Natl Res Council Canada, Inst Plant Biotechnol, Saskatoon, SK S7N 0W9, Canada
关键词
plant transformation; Agrobacterium tumefaciens; pea (Pisum sativum); genotype independence; GUS (beta-glucuronidase);
D O I
10.1016/S0168-9452(99)00267-8
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Transgenic pea plants can be produced by Agrobacterium-mediated transformation of thin slices from developing embryo axes. To determine if the method is effective for different pea genotypes, seven pea breeding lines adapted to western Canadian growing conditions were tested. using three different Agrobacterium tumefaciens transformation vectors. All vectors contained the gus (uidA) gene coding for the beta-glucuronidase (GUS) protein, but with different chemical selection genes. In total, 323 transgenic plants were recovered from 39 independent transformation events. Transgenic plants were recovered from each genotype and each selection system. but not from all combinations. GUS-positive explants were obtained from seeds harvested between 24 and 31 days after flowering. The mean time from Agrobacterium treatment to planting into soil averaged 186 days. Based on the initial number of seeds used, the transformation frequency was 0.6% (i.e. six independent transgenic events per 1000 axes sliced). The inserted genes were functional and inherited in a Mendelian fashion. Although more plants were recovered by selection on chlorsulfuron, GUS activity was generally greater in plants selected on kanamycin. GUS activity in the leaves of the original plants varied, but GUS activity in the second generation was correlated with that of the original transformants. (C) 2000 Elsevier Science Ireland Ltd. Ail rights reserved.
引用
收藏
页码:161 / 170
页数:10
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