草地早熟禾硝酸盐转运蛋白NRT2.4基因序列的克隆及表达分析  

Cloning and Expression Analysis of NRT2.4 Gene in Kentucky Bluegrass

作　　者：陈阳[1,2] 金一锋 金忠民[1] 王玉书 蔡荣建[1] 邵竹林[1] 王杰 CHEN Yang;JIN Yifeng;JIN Zhongmin;WANG Yushu;CAI Rongjian;SHAO Zhulin;WANG Jie(College of Life Science and Agro-Forestry,Qiqihar University,Qiqihar 161006,China;Heilongjiang Province Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas, Qiqihar 161006,China)

机构地区：[1]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006 [2]黑龙江省抗性基因工程与寒地生物多样性保护重点实验室,黑龙江齐齐哈尔161006

出　　处：《华北农学报》2021年第2期1-8,共8页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31701958);黑龙江省自然科学基金项目(QC2017026);黑龙江省普通高等学校青年创新人才培养计划项目(UNPYSCT-2018102);黑龙江省教育厅基本科研业务费科研项目(135309365);齐齐哈尔大学大学生创新创业项目(202010232161;202010232170)。

摘　　要：植物硝酸盐转运蛋白(Nitrate transporter,NRT)可有效调节与转运NO-3,提升氮素利用效率。分析草地早熟禾硝酸盐转运蛋白NRT2.4基因在不同氮素浓度下的表达规律,旨在揭示硝酸盐转运蛋白NRT2.4基因在氮素调控中扮演的角色。以草地早熟禾为材料,首先设计特异引物,利用RT-PCR技术克隆得到草地早熟禾NRT2.4基因cDNA序列,其长度为1694 bp,包含一个1257 bp的CDS区,编码418个氨基酸序列,属于Nitrate/nitrite transporter NarK超级家族1,与二穗短柄草、节节麦NRT2.4基因高度同源。第6,7个跨膜区之间有一段MFS家族所特有的序列特征:G-X3-D-X2-G-X-R,第4个跨膜区有典型的NO-3/NO-2转运载体特征序列:A-G-W-G/A-N-M-G,第8个跨膜区之后含有保守的蛋白激酶C识别基序(S/T-X-R/K):SKR。利用qRT-PCR方法,测定草地早熟禾NRT2.4基因表达水平存在组织特异性,叶部表达量最多,硝态氮利于NRT2.4基因的表达,氮饥饿和高浓度NaNO3水培液处理均利于NRT2.4基因的表达。上述结果为草地早熟禾硝酸盐转运蛋白NRT2.4在草坪草抗逆基因工程中的应用提供理论依据。Nitrate transporter(NRT)in plants can effectively regulate and transport NO-3 and improve nitrogen utilization efficiency.In order to investigate the relationship between Poa pratensis L.NRT2.4 and nitrogen stress,Poa pratensis L.was selected as the material for the cloning and bioinformatics analysis of NRT2.4 gene,and the regulation of this gene expression was observed under treatment by nitrogen nutrient solution.The cloned NRT2.4 gene of Poa pratensis L.belonged to the Nitrate/nitrite transporter NarK superfamily,and was highly homologous to that of Brachypodium distachyon(L.)Beauv.and Aegilops tauschii Coss.Between the 6th and 7th transmembrane domain(TMD),there was a sequence with signature of G-X3-D-X2-G-X-R unique to the MFS family.The 4th TMD had a typical NO-3/NO-2 transporter signature sequence of A-G-W-G/A-N-M-G.There was a conservative protein kinase C motif(S/T-X-R/K):SKR after the 8th TMD.The expression level of Poa pratensis L.NRT2.4 gene was tissue-specific,with the most expressed in leaves.Nitrate nitrogen was conducive to the expression of NRT2.4 gene,both nitrogen starvation and the treatment by nitrogen nutrient solution with a high NaNO3 were beneficial to the expression of NRT2.4 gene.The results above have provided a theoretical bad for the application of Poa pratensis L.NRT2.4 in stress-tolerance gene engineering in turfgrass.

关 键 词：草坪草 草地早熟禾 NRT2.4 氮素营养胁迫 克隆 表达 

分 类 号：Q78[生物学—分子生物学]