Programa de Pós-Graduação em Genética e Biologia Molecular, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
Plant Mol Biol. 2012 May;79(1-2):75-87. doi: 10.1007/s11103-012-9894-1. Epub 2012 Mar 1.
The soybean ubiquitous urease (encoded by GmEu4) is responsible for recycling metabolically derived urea. Additional biological roles have been demonstrated for plant ureases, notably in toxicity to other organisms. However, urease enzymatic activity is not related to its toxicity. The role of GmEu4 in soybean susceptibility to fungi was investigated in this study. A differential expression pattern of GmEu4 was observed in susceptible and resistant genotypes of soybeans over the course of a Phakopsora pachyrhizi infection, especially 24 h after infection. Twenty-nine adult, transgenic soybean plants, representing six independently transformed lines, were obtained. Although the initial aim of this study was to overexpress GmEu4, the transgenic plants exhibited GmEu4 co-suppression and decreased ureolytic activity. The growth of Rhizoctonia solani, Phomopsis sp., and Penicillium herguei in media containing a crude protein extract from either transgenic or non-transgenic leaves was evaluated. The fungal growth was higher in the protein extracts from transgenic urease-deprived plants than in extracts from non-transgenic controls. When infected by P. pachyrhizi uredospores, detached leaves of urease-deprived plants developed a significantly higher number of lesions, pustules and erupted pustules than leaves of non-transgenic plants containing normal levels of the enzyme. The results of the present work show that the soybean plants were more susceptible to fungi in the absence of urease. It was not possible to overexpress active GmEu4. For future work, overexpression of urease fungitoxic peptides could be attempted as an alternative approach.
大豆普遍存在的脲酶(由 GmEu4 编码)负责循环利用代谢产生的尿素。植物脲酶还具有其他生物学功能,特别是对其他生物体的毒性。然而,脲酶的酶活性与其毒性无关。本研究调查了 GmEu4 在大豆对真菌易感性中的作用。在接种 Phakopsora pachyrhizi 过程中,观察到易感性和抗性大豆基因型中 GmEu4 的差异表达模式,尤其是感染后 24 小时。获得了 29 株成年、转基因大豆植株,代表 6 个独立转化的系。尽管本研究的最初目的是过表达 GmEu4,但转基因植物表现出 GmEu4 的共抑制和脲酶活性降低。在含有转基因或非转基因叶片粗蛋白提取物的培养基中评估了 Rhizoctonia solani、Phomopsis sp. 和 Penicillium herguei 的生长情况。在缺乏脲酶的转基因植物的蛋白提取物中,真菌的生长高于非转基因对照的蛋白提取物。当感染 P. pachyrhizi 夏孢子时,缺乏脲酶的离体叶片比含有正常水平酶的非转基因叶片形成了更多数量的病斑、脓疱和爆发性脓疱。本工作的结果表明,在缺乏脲酶的情况下,大豆植物对真菌的敏感性更高。无法过表达有活性的 GmEu4。未来的工作可以尝试过表达具有抑菌活性的脲酶肽作为替代方法。