小麦条锈菌GMPP基因克隆及其产物糖转运功能分析  

Cloning and Sugar Transport Functional Analysis of GMPP in Puccinia striiformis f.sp.tritici

作　　者：刘秀峰 许静杨 杨兆顺[1] 楼辰军[1] 许高平 邵凤武[1] LIU Xiufeng;XU Jingyang;YANG Zhaoshun;LOU Chenjun;XU Gaoping;SHAO Fengwu(Institute of Crops Sciences,Tianjin Academy of Agricultural Sciences,Tianjin 300381,China;Institute of Plant Protection,Tianjin Academy of Agricultural Sciences,Tianjin 300381,China)

机构地区：[1]天津市农业科学院农作物研究所,天津300381 [2]天津市农业科学院植物保护研究所,天津300381

出　　处：《四川农业大学学报》2023年第1期1-5,53,共6页Journal of Sichuan Agricultural University

基　　金：天津市农业科学院青年科技创新项目(2020001);天津市应用基础与前沿技术研究计划(15JCYBJC30800)。

摘　　要：【目的】探讨条形柄锈菌小麦专化型真菌(Puccinia striiformis f.sp.tritici,Pst)中预测编码甘露糖-1-磷酸鸟苷酰转移酶(mannose-1-phosphate guanylyltransferase,GMPP)基因产物的糖转运功能。【方法】基于Pst转录组数据与NCBI中17个Pst基因组比对结果设计引物,筛选和克隆Pst的GMPP基因,命名为PsMT1,构建表达载体并导入酵母己糖转运缺失突变体EBY.VW4000,对获得的pDR196-PsMT1-EBY.VW4000重组酵母菌进行糖吸收功能互补测定。【结果】PsMT1基因的编码区为1245 bp,编码414个氨基酸,等电点为6.05。构建的无根进化树显示PsMT1与小麦秆锈菌的GMPP单独聚为一个分支。在酵母己糖转运功能缺失突变体中表达PsMT1能恢复EBY.VW4000菌体在D-甘露糖为唯一碳源的固体培养基上生长。【结论】克隆了Pst的PsMT1基因,其异源表达产物具有转运甘露糖的功能,为研究糖转运蛋白介导Pst-小麦互作的机制奠定基础。【Objective】This study was aimed to investigate the sugar transport function of mannose-1-phosphate guanyltransferase(GMPP)in Puccinia striiformis f.sp.tritici(Pst).【Method】Primers were designed to screen and amplify the GMPP gene(named PsMT1)based on the comparison of Pst transcriptome data with 17 Pst genomes sets in NCBI database,and the GMPP gene of Pst was cloned,named PsMT1.PsMT1 was expressed in the Saccharomyces cerevisiae mutant strain EBY.VW4000 and grew on media with hexose or sucrose as the sole carbon source.【Result】The cDNA of PsMT1 was 1245 bp in length,encoding 414 amino acids,and the isoelectric point was 6.05.Phylogenetic analysis indicated that PsMT1 had the closest genetic relationship with GMPP of P.graminis f.sp.tritici.Heterologous expression of PsMT1 can restore the growth of the hexose transporter deficient mutant yeast EBY.VW4000on medium with D-mannose as the sole carbon source.【Conclusion】PsMT1 was cloned and its heterologous expression product could transport mannose.The results obtained in this study can provide basis for the further study of interaction between Pst and wheat mediated by sugar transporter.

关 键 词：小麦条锈菌 糖转运 甘露糖-1-磷酸鸟苷酰转移酶 异源表达 

分 类 号：S435.121.42[农业科学—农业昆虫与害虫防治]