Majhi Bharat Bhusan, Shah Jasmine M, Veluthambi Karuppannan
Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625 021, Tamil Nadu, India.
Plant Cell Rep. 2014 Jun;33(6):929-44. doi: 10.1007/s00299-014-1572-0. Epub 2014 Feb 1.
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.
雄性不育转基因水稻植株TC-19在8号染色体上有一个新的T-DNA整合,伴有3号染色体6.55 kb片段和9号染色体29.8 kb片段的两个染色体间易位。我们报道了水稻(Oryza sativa)中一种与两个染色体间易位相关的复杂农杆菌T-DNA整合。携带单拷贝T-DNA的T-DNA标签水稻突变体TC-19在纯合条件下表现出雄性不育表型。通过基因组步移分析T-DNA末端与水稻基因组之间的连接,结果表明右边界侧翼为3号染色体序列,左边界侧翼为9号染色体序列。在3号染色体上进一步步移时,检测到3号染色体/8号染色体融合。从8号染色体断点的另一端进行基因组步移,发现了8号染色体/9号染色体融合。我们的研究结果表明,T-DNA连同3号染色体的6.55 kb区域和9号染色体的29.8 kb区域一起易位到了8号染色体上。对纯合TC-19突变体的Southern杂交分析表明,3号和9号染色体的天然序列得以恢复,但8号染色体在基因Os08g0152500的第一个内含子中的破坏并未恢复。复杂T-DNA整合到8号染色体导致了雄性不育。