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水稻中一种涉及两个染色体间易位的新型T-DNA整合。

A novel T-DNA integration in rice involving two interchromosomal translocations.

作者信息

Majhi Bharat Bhusan, Shah Jasmine M, Veluthambi Karuppannan

机构信息

Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, 625 021, Tamil Nadu, India.

出版信息

Plant Cell Rep. 2014 Jun;33(6):929-44. doi: 10.1007/s00299-014-1572-0. Epub 2014 Feb 1.

Abstract

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.

摘要

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。

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