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葡萄糖氧化酶催化分子氧氧化b -D葡萄糖产生葡萄糖酸和H2 O2 。产生H2 O2 和其它活性氧物质是植物对病原体侵入的防御反应。实验证明H2 O2 水平升高不仅可诱发细胞过敏坏死反应 ,而且还能激活抗病基因表达如植物抗毒素及诱导病原相关蛋白 (PR)等基因的表达。通过农杆菌介导将来自黑麴霉(Aspergillusniger)的葡萄糖氧化酶基因转入 2个马铃薯重要栽培品种大西洋 (Atlantic)和夏普蒂 (Shepody)中获得 2 3个转基因株系 ,其中 75 %为Kan抗性植株。在 2 3个转基因株系中有 16个株系可用PCR扩增出目的基因。进一步用核酸斑点杂交选出 13个杂交阳性株系。转基因植物总DNA的Southernblot分析表明 ,GO基因已整合到马铃薯四倍体基因组中 ,转基因大西洋植株中目的基因为 2～ 4个拷贝 ,而夏普蒂转基因植株中为 1个拷贝。将转基因植株离体叶片接种呼和浩特地区流行的Phytophthoroinfestans的复合生理小种 1,3,4孢子。结果表明 ,和未转基因对照相比转基因植株病斑出现时间晚 ,病斑扩展速度慢 ,发病程度轻 ,感病叶片少。总体上表现出较明显抗病性。转基因大西洋中抗性株系多于夏普蒂。转基因马铃薯植株体内H2 O2 含量测定表明 ,转基因马铃薯植株的根系和叶片在KI-淀粉培养基上数天后变为蓝色 ,而未转基因对照植株的根系?
Glucose oxidase catalyzes the molecular oxygen oxidation of b-D glucose to produce gluconic acid and H2 O2. The production of H2O2 and other reactive oxygen species is a defensive response of plants to pathogen intrusion. Experimental results show that elevated H2O2 can not only induce cellular allergic necrosis reaction, but also activate the expression of disease resistance genes such as phytoalexin and induced pathogen-related protein (PR) and other genes. Agrobacterium-mediated transformation of glucose oxidase gene from Aspergillus niger into two potato cultivars Atlantic and Shepody obtained 23 transgenic lines, of which 75% were Kan Resistant plants. 16 of the 23 transgenic lines amplified the target gene by PCR. Further hybridization with nucleic acid dot selected 13 hybrid positive lines. Southern blot analysis of the total DNA of transgenic plants showed that the GO gene has been integrated into the tetraploid potato genome with 2 to 4 copies of the target gene in transgenic Atlantic plants and 1 copy in the Shatiopi transgenic plants. The transgenic plants were inoculated with the leaves of 1,3,4 spores of the composite physiological races of Phytophthoroinfestans endemic in Hohhot. The results showed that compared with the non-transgenic control, the transgenic plants appeared late, the lesion extended slowly, the incidence was light and the susceptible leaves were few. Overall showed more obvious disease resistance. Transgenic Atlantic strain more than Sharpe. Determination of H2O2 content in transgenic potato plants showed that the roots and leaves of transgenic potato plants turned blue in KI-starch medium several days later, while the roots of non-transgenic control plants were?