Xuan Yuan Hu, Peterson Thomas, Han Chang-Deok
College of Plant Protection, Shenyang Agricultural University, Dongling Road 120, Shenyang, Liaoning, 110866, China.
Division of Applied Life Science (BK21 Program), Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju, 660-701, Korea.
Methods Mol Biol. 2016;1469:49-61. doi: 10.1007/978-1-4939-4931-1_4.
Closely-located transposable elements (TEs) have been known to induce chromosomal breakage and rearrangements via alternative transposition. To study genome rearrangements in rice, an Ac/Ds system has been employed. This system comprises an immobile Ac element expressed under the control of CaMV 35S promoter, and a modified Ds element. A starter line carried Ac and a single copy of Ds at the OsRLG5 (Oryza sativa receptor-like gene 5). To enhance the transpositional activity, seed-derived calli were cultured and regenerated into plants. Among 270 lines regenerated from the starter, one line was selected that contained a pair of inversely-oriented Ds elements at the OsRLG5 (Oryza sativa receptor-like gene 5). The selected line was again subjected to tissue culture to obtain a regenerant population. Among 300 regenerated plants, 107 (36 %) contained chromosomal rearrangements including deletions, duplications, and inversions of various sizes. From 34 plants, transposition mechanisms leading to such genomic rearrangements were analyzed. The rearrangements were induced by sister chromatid transposition (SCT), homologous recombination (HR), and single chromatid transposition (SLCT). Among them, 22 events (65 %) were found to be transmitted to the next generation. These results demonstrate a great potential of tissue culture regeneration and the Ac/Ds system in understanding alternative transposition mechanisms and in developing chromosome engineering in plants.
已知紧密定位的转座元件(TEs)可通过交替转座诱导染色体断裂和重排。为了研究水稻中的基因组重排,采用了Ac/Ds系统。该系统由一个在CaMV 35S启动子控制下表达的非移动性Ac元件和一个修饰的Ds元件组成。一个起始系在水稻类受体基因5(OsRLG5)处携带Ac和单拷贝的Ds。为了增强转座活性,将种子来源的愈伤组织进行培养并再生为植株。在从起始系再生的270个株系中,选择了一个在OsRLG5处含有一对反向排列的Ds元件的株系。将所选株系再次进行组织培养以获得再生群体。在300株再生植株中,107株(36%)含有包括各种大小的缺失、重复和倒位在内的染色体重排。对34株植株分析了导致这种基因组重排的转座机制。重排是由姐妹染色单体转座(SCT)、同源重组(HR)和单染色单体转座(SLCT)诱导的。其中,22个事件(65%)被发现可传递给下一代。这些结果证明了组织培养再生和Ac/Ds系统在理解交替转座机制以及植物染色体工程开发方面具有巨大潜力。