A novel T-DNA integration in rice involving two interchromosomal translocations
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A male sterile transgenic rice plant TC-19 harboured a novel T-DNA integration in chromosome 8 with two interchromosomal translocations of 6.55 kb chromosome 3 and 29.8 kb chromosome 9 segments.
We report a complex Agrobacterium T-DNA integration in rice (Oryza sativa) associated with two interchromosomal translocations. The T-DNA-tagged rice mutant TC-19, which harboured a single copy of the T-DNA, displayed male sterile phenotype in the homozygous condition. Analysis of the junctions between the T-DNA ends and the rice genome by genome walking showed that the right border is flanked by a chromosome 3 sequence and the left border is flanked by a chromosome 9 sequence. Upon further walking on chromosome 3, a chromosome 3/chromosome 8 fusion was detected. Genome walking from the opposite end of the chromosome 8 break point revealed a chromosome 8/chromosome 9 fusion. Our findings revealed that the T-DNA, together with a 6.55-kb region of chromosome 3 and a 29.8-kb region of chromosome 9, was translocated to chromosome 8. Southern blot analysis of the homozygous TC-19 mutant revealed that the native sequences of chromosome 3 and 9 were restored but the disruption of chromosome 8 in the first intron of the gene Os08g0152500 was not restored. The integration of the complex T-DNA in chromosome 8 caused male sterility.
KeywordsAgrobacterium Genome walking Interchromosomal translocation Male sterility Rice T-DNA integration
Murashige and Skoog
Hygromycin phosphotransferase gene
Chitinase gene of Trichoderma virens
Homozygous transgenic event
Hemizygous transgenic event
T-DNA left border
T-DNA right border
Basic local alignment search tool
International rice genome sequencing project
National Center for Biotechnology Information
We thank Dr. K. Dharmalingam, School of Biotechnology, Madurai Kamaraj University for his permission to use the radioisotope facility. This work was supported by Department of Biotechnology, Ministry of Science & Technology, Government of India [Project entitled “Functional Analysis of Gene Regulatory Networks During Flower and Seed development in rice”, Project No. BT/AB/FG-I(PH-II)(5)2009].
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