Pastori Gabriela M, Huttly Alison, West Jevon, Sparks Caroline, Pieters Alejandro, Luna Celina M, Jones Huw D, Foyer Christine H
Crop Performance and Improvement Division, Rothamsted Research, Harpenden, Hertfordshire AL5 2JQ, UK.
IVI, Centro de Ecología, Altos de Pipe, Carretera Panamericana Km 11, Apartado 21827, Caracas 1020-A, Venezuela.
Funct Plant Biol. 2007 Sep;34(9):835-843. doi: 10.1071/FP07112.
The aim of the present study was to provide useful background information and evidence of the functionality of the maize Activator/Dissociation (Ac/Ds) system in hexaploid wheat. Two transgenic parental wheat lines, one harbouring the immobilised Ac element (iAc) and the other the Ds element (pUbi[Ds-uidA]bar), were crossed. Transient GUS assays confirmed that the iAc transposase is active in hexaploid wheat. Selected F1 and F2 lines were analysed by PCR using primers specific to Ac, uidA and bar genes. The primer pair Ubi/bar-tag was used to detect excision of the Ds-uidA sequence, which occurred at a frequency of 39% in the F1 generation. Lines free of Ac and showing evidence of Ds excision were subject to Southern analysis, which indicated that at least one transposition event might have occurred in these lines. Although more evidence is required to unequivocally support the reintegration of the Ds element in the wheat genome, the evidence presented here nevertheless demonstrates the effectiveness and potential value of using this system to tag genes in wheat.
本研究的目的是提供关于玉米激活子/解离子（Ac/Ds）系统在六倍体小麦中功能的有用背景信息和证据。将两个转基因亲本小麦品系进行杂交，一个含有固定化的Ac元件（iAc），另一个含有Ds元件（pUbi[Ds-uidA]bar）。瞬时GUS分析证实iAc转座酶在六倍体小麦中具有活性。使用针对Ac、uidA和bar基因的特异性引物，通过PCR对选定的F1和F2品系进行分析。引物对Ubi/bar-tag用于检测Ds-uidA序列的切除，该序列在F1代中的切除频率为39%。不含Ac且显示Ds切除证据的品系进行了Southern分析，结果表明这些品系中可能至少发生了一次转座事件。尽管需要更多证据来明确支持Ds元件在小麦基因组中的重新整合，但本文提供的证据证明了使用该系统在小麦中标记基因的有效性和潜在价值。
