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

被引：0
作者：
Fei-Fei Li
Shen-Jie Wu
Tian-Zi Chen
Jie Zhang
Hai-Hai Wang
Wang-Zhen Guo
Tian-Zhen Zhang
机构：
[1] Nanjing Agricultural University,National Key Laboratory of Crop Genetics & Germplasm Enhancement
[2] Shanxi Academy of Agricultural Science,Cotton Research Institute
来源：
Plant Cell, Tissue and Organ Culture (PCTOC) | 2009年 / 97卷
关键词：
Upload cotton; Co-transformation; Transformation frequency;
D O I：
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中图分类号：
学科分类号：
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 T1 plants demonstrated that multiple transgenes were inherited and expressed in progenies.
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页码：225 / 235
页数：10
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