Transformation of the obligate biotrophic rust fungus Uromyces fabae

专性生物营养型锈菌豆尿霉的转化

• 批准号: 36044083
• 负责人: Professor Dr. Ralf Thomas Vögele
• 金额: --
• 依托单位: Fachgebiet Phytopathologie (360a)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/36044083?language=en
• 关键词: Transformation; obligate; biotrophic; rust; fungus

Obligate biotrophic fungi, like the rust fungi, represent a major class of pathogens especially of economically important crops world wide. Yet, research on rust fungi and other obligate biotrophs has been hampered by the absence of a system for stable transformation. We have established Biolistics as a delivery tool for transgenes into the legume rust Uromyces fabae. Endogenous regulatory elements derived from the plasma membrane ATPase gene (Uf- PMA1) are used to drive expression of color and selection markers. GUS, GFP and DsRed have been established as color markers, while a mutated form of the endogenous gene succinate dehydrogenase 1 (SucDH1(H254Y)) is used as selection marker. The system uses the fungicide Carboxin as selective agent and allows a selection of transformants directly on the host plant Vicia faba. However, so far the transgene is lost after a couple of rounds of propagation. We are in the process of establishing a second transformation method based on Agrobacterium tumefaciens mediated gene delivery. This system uses the same control elements, markers and selection procedure as the biolistic system, but has the advantage of an increased stability of the transgenes. Future work will therefore center on the improvement of the A. tumefaciens mediated technique and on the extension of the method to other rust fungi.

专性生物营养真菌，如锈菌，代表了一类主要的病原体，特别是在世界范围内重要的经济作物。然而，由于缺乏稳定转化系统，对锈菌和其他专性生物营养菌的研究一直受到阻碍。我们已经建立了生物学作为转基因进入豆科锈菌的传递工具。来自质膜atp酶基因（Uf- PMA1）的内源性调控元件用于驱动颜色和选择标记的表达。建立了GUS、GFP和DsRed作为颜色标记，内源基因琥珀酸脱氢酶1的突变形式（SucDH1(H254Y)）作为选择标记。该系统使用杀菌剂Carboxin作为选择剂，并允许直接在宿主植物蚕豆上选择转化体。然而，到目前为止，转基因在几轮繁殖后就丢失了。我们正在建立基于农杆菌介导的基因传递的第二种转化方法。该系统使用与生物系统相同的控制元件、标记和选择程序，但其优点是增加了转基因的稳定性。因此，未来的工作将集中在改进肿瘤分枝杆菌介导技术和将该方法推广到其他锈菌上。