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土壤中锌的缺乏已成为农业中普遍存在的问题,会导致粮食的减产减质。ZIP家族蛋白质对农作物吸收Zn与Fe起着关键作用。本研究利用RACE技术从豆科植物紫云英中分离到一个全长的ZIP家族Zn转运基因,命名为AsZIP2;利用热不对称交错PCR与反向PCR方法获取了AsZIP2基因上游长度为1.6 kb的启动子序列。生物信息学分析表明AsZIP2基因编码339个氨基酸;预测的AsZIP2蛋白质含有保守的ZIP结构域并由9个跨膜结构域构成;序列比对与系统进化分析表明AsZIP2与蒺藜苜蓿及日本百脉根的Zn转运蛋白ZIP2的亲缘关系密切。利用RT-PCR与定量PCR方法检测紫云英AsZIP2基因在丛枝菌根中的表达,结果显示AM真菌的侵染强烈地抑制AsZIP2的表达。与已知的Mt ZIP2基因一致,施加Zn会诱导AsZIP2的表达。有趣地是,在低磷条件下,AsZIP2在根中的表达显著地增强。研究结果表明,AM真菌,Zn或Pi水平均影响Zn转运基因AsZIP2在根中的表达水平。对AsZIP2基因在分子特征与表达谱方面的初步研究将有利于进一步功能性鉴定该基因参与植物生长和发育。
The lack of zinc in the soil has become a common problem in agriculture, will lead to the reduction of grain production and quality. ZIP family proteins absorb Zn and Fe from crops play a key role. In this study, a full-length ZIP family Zn transport gene was isolated from the leguminous plant Vetchum by using RACE technology and named as AsZIP2. The upstream of AsZIP2 gene was obtained by thermal asymmetric staggered PCR and reverse PCR. Promoter sequence. Bioinformatics analysis showed that the AsZIP2 gene encoded 339 amino acids. The predicted AsZIP2 protein contained a conserved ZIP domain and consisted of nine transmembrane domains. Sequence alignment and phylogenetic analysis showed that AsZIP2 was positively associated with Medicago truncatula and Japanese Lotus roots Zn transporter ZIP2 is closely related. The expression of AsZIP2 gene in arbuscular mycorrhizas was detected by RT-PCR and quantitative PCR. The results showed that AM fungal infection strongly inhibited AsZIP2 expression. Consistent with the known Mt ZIP2 gene, Zn was administered to induce AsZIP2 expression. Interestingly, under low phosphorus conditions, expression of AsZIP2 in the root is significantly enhanced. The results showed that AM fungi, Zn or Pi levels affected the expression of Zn transport gene AsZIP2 in roots. A preliminary study on the molecular characteristics and expression profile of AsZIP2 gene will facilitate further functional identification of this gene involved in plant growth and development.