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Agrobacterium-mediated co-transformation of multiple genes in upland cotton

文献类型： 外文期刊

作者： Li, Fei-Fei 1 ; Wu, Shen-Jie 1 ; Chen, Tian-Zi 1 ; Zhang, Jie 1 ; Wang, Hai-Hai 1 ; Guo, Wang-Zhen 1 ; Zhang, Tian-Zhe 1 ;

作者机构： 1.Nanjing Agr Univ, Natl Key Lab Crop Genet & Germplasm Enhancement, Nanjing 210095, Peoples R China

2.Shanxi Acad Agr Sci, Cotton Res Inst, Yuncheng 044000, Peoples R China

关键词： Upload cotton;Agrobacterium tumefaciens;Co-transformation;Transformation frequency

期刊名称：PLANT CELL TISSUE AND ORGAN CULTURE （ 影响因子：2.711； 五年影响因子：2.73 ）

ISSN： 0167-6857

年卷期： 2009 年 97 卷 3 期

页码：

收录情况： SCI

摘要： Two cotton genotypes, Simian 3 (SM 3) and WC, were co-transformed using a mixture of four Agrobacterium tumefaciens cultures of strain LBA4404, each carrying a plasmid harboring the following genes, Bt + sck (for Bacillus thuringenesis protein and modified Cowpea trypsin inhibitor), bar (for glufosinate), keratin, and fibroin. The frequency of callus induction, embryogenesis, and plant regeneration were notably different between the two genotypes. However, there were no differences between the two genotypes for number of plantlets carrying multiple gene copies of different gene combinations as well as transformation frequency for different gene combinations. PCR analysis indicated that more than 80% of plantlets carried the nptII gene for kanamycin resistance. Overall, the co-transformation frequency of two or more genes was about 35%. Southern blot analysis confirmed integration of target genes into the cotton genome, and the number of copies of the transgene(s) varied from one to four. Multiple transgene expression was confirmed by RT-PCR analysis in some transgenic lines. Further analysis of T(1) plants demonstrated that multiple transgenes were inherited and expressed in progenies.

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