中外葡萄与葡萄酒Issue(5)：1-9,9.DOI:10.13414/j.cnki.zwpp.2025.05.001
酿酒葡萄铵转运体AMT1;1基因的克隆与功能分析
Cloning and Function Determination of Ammonium Transporter Gene AMT1;1 in Wine Grape
摘要
Abstract
An ammonium(NH4+)transporter-encoding gene VvAMT1;1 was cloned from the wine grape'Marselan'.VvAMT1;1 shares 76.10%amino acid sequence identity with homologs from peach PpeAMT1;1,pear PbrAMT1;1,rice OsAMT1;1,and Arabidopsis AtAMT1;1,AtAMT1;2,and AtAMT1;3.Phylogenetic analysis revealed that VvAMT1;1 is evolutionarily closer to peach AMT1;1.The expression level of VvAMT1;1 varied significantly across different developmental stages of'Marselan'fruit.It was highly expressed during the young fruit stage,hard core stage,and veraison stage,but significantly decreased during the second expansion stage and ripening stage.Although the expression levels in annual new leaves and roots were lower than in fruits at the same stage,they remained relatively stable throughout the entire fruit developmental period.The expression level of the VvAMT1;1 gene varied among fruits from different graft combinations,which was significantly higher in the'Marselan/SO4'graft combination,compared to the'Marselan/1103P'and own-rooted'Marselan'grapevines.Yeast mutant functional complementation and 15N isotope labeled uptake kinetics analysis demonstrated that VvAMT1;1 is a typical high-affinity ammonium transporter,with a Km value of 35.67 μmol·L-1 and an Vmax value of 2.85 μmoles·min-1·μg-1 cells,dominates the NH4+uptake in yeast cells.Moreover,methylamine(MeA+)significantly reduced the Vmax value for VvAMT1;1-mediated NH4+uptake,and VvAMT1;1 can mediate the weak uptake and utilization of MeA+in yeast cells.L-methionine sulfoximine(L-MSX)significantly reduced the Vmax value for VvAMT1;1-mediated NH4+uptake.The VvAMT1;1-mediated NH4+uptake process was subject to feedback inhibition by endogenous NH4+accumulation levels.关键词
酿酒葡萄/铵转运体/果实发育/酵母突变体功能互补/底物反馈抑制Key words
wine grape/ammonium transporter/fruit development/functional complementation in yeast mutant/substrate feedback inhibition分类
农业科技引用本文复制引用
曾蕊,朱骏驰,徐俐琴,刘秀云,韩菲菲,唐美玲,唐艳雯,宋志忠..酿酒葡萄铵转运体AMT1;1基因的克隆与功能分析[J].中外葡萄与葡萄酒,2025,(5):1-9,9.基金项目
上海市晨光计划基金项目(KY4-0100-20-01) （KY4-0100-20-01）
山东省重点研发计划(重大科技创新工程)(2022CXGC010605)国家留学基金(202208370080) （重大科技创新工程）
