Integration and characterization of T-DNA insertion in upland cotton
2013
Yang, X., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Li. F., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Zhang, X., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Liu, K., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Wang, Q., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Zhang, Ch., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Liu, Ch., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Zhu, W., Henan Agricultural Univ., Zhengzhou (China). Agronomy Coll. | Shan, G., Chinese Academy of Agricultural Sciences, Anyang (China). Cotton Research Inst. | Chin, Ch.K., Rutgers Univ., New Jersey (USA). Dept. of Plant Science | Fang, W., Henan Academy of Agricultural Sciences, Zhengzhou (China). Economic Crop Research Inst.
Copy numbers were evaluated by real-time quantitative PCR and 149 junctions of T-DNA were isolated by thermal asymmetric interlaced PCR from 92 independent transgenic cotton lines transformed by Agrobacterium tumefaciens strain LBA4404. Real-time quantitative PCR results showed that 46% had integration of one or two T-DNA copies, 54% had three or more copies. Among 63 amplified products at LB junctions, 51% showed co-transformation of the vector backbone, 30% retained a portion of LB ranging from 3 to 23 bp, and 19% showed deletions ranging from 1 to 148 bp from the LB inner end. In contrast, all of the cleavage sites were located in the inner region of RB. The distribution of T-DNA insertions in upland cotton genome included coding sequences, transposons, plastid-derived sequences and microsatellites.
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