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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。
Abstract：Zinc deficiency in soil which has led to the reduction of grain yield and quality has become a common problem in agriculture. The ZRT-IRT-like protein(ZIP) plays a pivotal role in the acquisition of Zn and Fe in crop plants. In this study,a full length c DNA of ZIP family of transporter encoding gene,designed as AsZIP2,was isolated from legume Astragalus sinicus through RACE technique;and a 1.6 kb length of promoter sequence of AsZIP2 gene was identified using TAIL-PCR and inverse PCR methods. In silico analysis showed that AsZIP2 encoded 339 aa protein;the deduced AsZIP2 protein which was composed of 9 transmembrane domains contains a conserved ZIP domain. Sequence alignment and phylogenetic analysis indicated that AsZIP2 has close genetic relationship with Medicago truncatula and Lotus japonicus Zn transporter ZIP2. The RT-PCR and quantitative PCR methods were applied to detect the expression of Chinese milk vetch AsZIP2 gene in arbuscular mycorrhizal roots.The result showed that the infection of AM fungi strongly repressed the expression of AsZIP2. In agreement with the Mt ZIP2 gene identified,AsZIP2 was induced in roots by Zn fertilization. Interestingly,the AsZIP2 gene was dramatically enhanced in roots under low Pi conditions. These results indicated that the expression levels of AsZIP2 gene in roots was influenced by the AM fungi,Zn or Pi status. The initial study of AsZIP2 gene in the aspects of molecular characteristics and expression profiles will be useful for further functional identification of the gene involving in the growth and development of A. sinicus.
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基本信息:
DOI：10.13417/j.gab.035.000610
中图分类号:S541.3;S182
引用信息:
[1]韩亚超,谢贤安,范晓宁,等.紫云英丛枝菌根共生锌转运蛋白基因AsZIP2的克隆与表达调控[J].基因组学与应用生物学,2016,35(03):610-621.DOI:10.13417/j.gab.035.000610.
基金信息:
国家自然科学基金项目“丛枝菌根真菌Glomus mosseae Gm201基因在共生早期信号转导中的生物学功能研究”(31270159);; 教育部高等学校青年骨干教师国内访问学者项目(201312045);; 安徽省教育厅振兴计划“地方高职院校构建产学研平台的研究与实践”(2014ZDJY120)和安徽省教育厅自然科学项目“生态农业模式下淮北地区沼液中微生物多样性分析及土壤修复性能的研究”(KJ2015A365)共同资助
2016-03-25
2016-03-25