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[目的]为了解决大麦根组织与根际微生物(有益菌或致病菌)互作分析系统的缺失,建立了大麦根部快速稳定遗传转化系统。[方法]显微镜下通过分离大麦未成熟幼胚并剥离胚轴,对大麦幼胚实施含有外源基因GFP、Hv EXPANSINB1-RNAi、BAX INHIBITOR-1以及GFP-ER的农杆菌转化,将大麦幼胚分别转入愈伤培养基和生根培养基进行根组织的诱导。[结果]大量的根可以在6 w时间内被成功诱导。利用该体系:实现GFP在根中表达;转有Hv EXPANSINB1-RNAi基因的根毛数量显著减少;经BAX INHIBITOR-1基因转化的根组织减少了病原菌Fusarium graminearum的定殖;成功地检测到了GFP-ER基因在内质网及细胞核周围的定殖。[结论]该体系可成功应用于外源基因的表达、沉默,大麦根部功能基因与病原菌的互作以及蛋白的亚细胞定位研究。
[Objective] The purpose of this research was to establish a rapid and stable genetic transformation system of barley root in order to solve the lack of interaction analysis system between rhizobium and rhizosphere microbes (beneficial bacteria or pathogenic bacteria). [Method] Agrobacterium tumefaciens transformed with exogenous genes GFP, Hv EXPANSINB1-RNAi, BAX INHIBITOR-1 and GFP-ER were transformed into immature embryos of barley by using immature embryos isolated from the barley under microscope. Were transferred to the callus medium and rooting medium for root tissue induction. [Result] A large number of roots could be successfully induced within 6 w time. The number of root hairs transfected with Hv EXPANSINB1-RNAi gene was significantly reduced. The root tissues transformed with BAX INHIBITOR-1 gene reduced the colonization of Fusarium graminearum. The results of GFP-ER Genes are colonized in the endoplasmic reticulum and nucleus. [Conclusion] The system could be successfully applied to the expression of exogenous genes, silencing, the interaction between functional genes of root of barley and pathogens, and the subcellular localization of proteins.