<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated co-transformation of multiple genes in upland cotton |
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Authors: | Fei-Fei Li Shen-Jie Wu Tian-Zi Chen Jie Zhang Hai-Hai Wang Wang-Zhen Guo Tian-Zhen Zhang |
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Institution: | (1) National Key Laboratory of Crop Genetics & Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China;(2) Cotton Research Institute, Shanxi Academy of Agricultural Science, Yuncheng, 044000, China |
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Abstract: | 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.
Fei-Fei Li and Shen-Jie Wu are joint first authors. |
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Keywords: | Upload cotton Agrobacterium tumefaciens Co-transformation Transformation frequency |
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