生命科学研究2019，Vol.23Issue(1)：1-12,12.DOI:10.16605/j.cnki.1007-7847.2019.01.001
异源表达芸薹生链格孢谷氨酸脱氢酶基因AbGDH提高水稻氮素利用率的研究
Ectopic Expression of a Fungal AbGDH Gene from Alternaria brassicicola Improves Nitrogen Use Efficiency in Rice
摘要
Abstract
Nitrogen is one of the most important elements in agriculture industry.NADP(H)-dependent glutamate dehydrogenase (GDH) has a lower affinity for ammonium in higher plants,whose nitrogen is thus mainly assimilated by the glutamine synthetase (GS)/glutamate synthase (GOGAT) pathway.But,in lower organ isms such as fungi,GDHs play an important role in nitrogen assimilation due to their higher affinity for ammonium.In this study,a glutamate dehydrogenase gene AbGDH from Alternaria brassicicola was cloned and successfully over-expressed in rice (Oryza sativa L.cv.Kitaake).Enzyme activity assay in vitro demonstrated that the Km values of AbGDH for NH4+,2-oxoglutarate and glutamate were 2.144±0.141 mmol/L,2.690± 0.233 mmol/L and 96.772±0.542 mmol/L,respectively.Meanwhile,the enzyme activity assay in vivo revealed that,compared with the nontransgenic plants,the AbGDH over-expressing plants showed a higher aminating activity and a lower Km value for NH4+.Additionally,the hydroponic experiments revealed that,compared with control plants,the transgenic seedlings exhibited a significant increase in plant height and dry weight at lower nitrogen conditions.These results suggested that ectopic expression of AbGDH was inclined to convert 2-oxoglutarate to glutamate,which subsequently improved the ammonium assimilation and enhanced the nitrogen use efficiency particularly under low nitrogen fertility.关键词
谷氨酸脱氢酶(GDH)/氮素利用率/水稻/芸薹生链格孢(Alternaria brassicicola)Key words
glutamate dehydrogenase (GDH)/nitrogen assimilation/rice/Alternaria brassicicola分类
农业科技引用本文复制引用
陈鸣冬,刘聪,刘德荣,汤琦龙,林建中,刘选明..异源表达芸薹生链格孢谷氨酸脱氢酶基因AbGDH提高水稻氮素利用率的研究[J].生命科学研究,2019,23(1):1-12,12.基金项目
国家自然科学基金资助项目(31571635,31871595) （31571635,31871595）
湖南省科技重大专项(2018NK1010) （2018NK1010）
湖南省自然科学基金资助项目(2017JJ2042) （2017JJ2042）
湖南省科技计划重点项目(2017WK2012) （2017WK2012）
长沙市科技计划项目(kq1801001,kq1701028) （kq1801001,kq1701028）
水稻生物学国家实验室开放课题(150103) The National Science Foundation of China (31571635,31871595) （150103）
Hunan Provincial Technology Major Special Projects (2018NK1010) （2018NK1010）
Hunan Provincial Natural Science Foundation of China (2017JJ2042) （2017JJ2042）
Planned Science and Technology Project of Hunan Province (2017WK2012) （2017WK2012）
Planned Science and Technology Project of Changsha City (kq1801001,kq1701028) （kq1801001,kq1701028）
the Public Subject of State Key Laboratory of Rice Biology (150103) （150103）
