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为探究外生菌根真菌对油松磷吸收作用的分子机理,以油松优良乡土外生菌根真菌——浅黄根须腹菌Rhizopogon luteolus的磷酸盐转运蛋白基因(Rl PT)为对象,在缺陷酵母MB192中进行了异源表达研究。结果显示,该c DNA编码的蛋白质能够互补高亲和力磷酸盐转运蛋白pho84的功能;由不同pH条件下生长试验可知,该蛋白是一个与质子相偶联的运输蛋白;Rl PT测算的K_m值为57.90μmol/L磷酸盐;通过酸性磷酸酶的活性检测,进一步验证该基因是具有高亲和力磷酸盐转运蛋白功能的基因。激光共聚焦显微观察表明,该蛋白在低磷条件下多定位于酵母细胞膜上发挥其功能。
In order to explore the molecular mechanism of ectomycorrhizal fungi on the absorption of Pinus tabulaeformis, we used the phosphate transporter gene (Rl PT) of Rhizopogon luteolus, an ectomycorrhizal fungi of Pinus tabulaeformis, Defective yeast MB192 in heterologous expression studies. The results showed that the c-DNA encoded protein could complement the function of high-affinity phosphate transporter pho84. According to the growth test under different pH conditions, the protein was a transport protein coupled with proton. The K_m value of Rl PT was 57.90μmol / L phosphate. The activity of acid phosphatase was tested to further verify that the gene is a high-affinity phosphate transporter. Laser confocal microscopy showed that the protein under the low phosphorus conditions in the yeast cell membrane to play its function.