Department of Genetics, Development and Cell Biology, Department of Agronomy, Iowa State University, Ames, IA 50011, USA.
Theor Appl Genet. 2012 Nov;125(7):1449-62. doi: 10.1007/s00122-012-1925-4. Epub 2012 Jul 14.
The maize Activator (Ac)/Dissociation (Ds) transposable element system has been used in a variety of plants for insertional mutagenesis. Ac/Ds elements can also generate genome rearrangements via alternative transposition reactions which involve the termini of closely linked transposons. Here, we introduced a transgene containing reverse-oriented Ac/Ds termini together with an Ac transposase gene into rice (Oryza sativa ssp. japonica cv. Nipponbare). Among the transgenic progeny, we identified and characterized 25 independent genome rearrangements at three different chromosomal loci. The rearrangements include chromosomal deletions and inversions and one translocation. Most of the deletions occurred within the T-DNA region, but two cases showed the loss of 72 kilobase pairs (kb) and 79 kb of rice genomic DNA flanking the transgene. In addition to deletions, we obtained chromosomal inversions ranging in size from less than 10 kb (within the transgene DNA) to over 1 million base pairs (Mb). For 11 inversions, we cloned and sequenced both inversion breakpoints; in all 11 cases, the inversion junctions contained the typical 8 base pairs (bp) Ac/Ds target site duplications, confirming their origin as transposition products. Together, our results indicate that alternative Ac/Ds transposition can be an efficient tool for functional genomics and chromosomal manipulation in rice.
玉米激活因子(Ac)/解离(Ds)可转座元件系统已被用于多种植物的插入诱变。Ac/Ds 元件还可以通过涉及紧密连锁转座子末端的替代转座反应产生基因组重排。在这里,我们将一个含有反向取向的 Ac/Ds 末端的转基因以及 Ac 转座酶基因导入水稻(Oryza sativa ssp. japonica cv. Nipponbare)。在转基因后代中,我们在三个不同的染色体位点鉴定和表征了 25 个独立的基因组重排。这些重排包括染色体缺失和倒位以及一个易位。大多数缺失发生在 T-DNA 区域内,但有两个情况显示缺失了 72 千碱基对(kb)和 79 kb 侧翼转基因的水稻基因组 DNA。除了缺失,我们还获得了大小从小于 10 kb(在转基因 DNA 内)到超过 100 万个碱基对(Mb)的染色体倒位。对于 11 个倒位,我们克隆并测序了两个倒位断点;在所有 11 个案例中,倒位连接处都包含典型的 8 个碱基对(bp)Ac/Ds 靶位点重复,证实了它们作为转座产物的起源。总之,我们的结果表明,替代 Ac/Ds 转座可以成为水稻功能基因组学和染色体操作的有效工具。