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引用本文:李迎,米铁柱,乔玲,甄毓.基于转录组测序对两种氮素营养条件下多形微眼藻氮代谢途径的解析.海洋与湖沼,2019,50(6):1241-1251.
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基于转录组测序对两种氮素营养条件下多形微眼藻氮代谢途径的解析
李迎1,2, 米铁柱1,3,2, 乔玲4, 甄毓1,3,2
1.海洋环境与生态教育部重点实验室 青岛 266100;2.中国海洋大学 环境科学与工程学院 青岛 266100;3.青岛海洋科学与技术试点国家实验室 海洋生态与环境科学功能实验室 青岛 266071;4.中国水产科学研究院黄海水产研究所 青岛 266071
摘要:
探究微藻的氮代谢通路对了解其对不同氮源利用的分子机制具有重要意义。本研究利用Illumina高通量测序技术对两种氮素营养条件下(硝酸氮和尿素)多形微眼藻的转录组进行分析,通过基因功能注释及数字基因表达谱分析,研究了多形微眼藻细胞内氮代谢的调控机制。结果检测出15种参与氮代谢的酶,对应76个编码基因,构建了多形微眼藻的氮代谢通路图。其中10个酶编码基因在两种不同氮素营养条件下存在差异表达,最显著的是谷氨酸合成酶、谷氨酸脱氢酶和谷氨酰胺合成酶相关基因。有机氮源(尿素)实验组中,多形微眼藻细胞内的硝酸盐还原酶、亚硝酸盐还原酶等基因的差异表达明显高于无机氮源(硝酸钠)实验组,表明当环境中的氮源为尿素时,会对多形微眼藻细胞内硝酸盐的转化和利用有一定影响。本研究初步阐述了硝酸盐、尿素的吸收转运对多形微眼藻细胞内氮代谢的影响机制,可为硅藻在不同氮素营养条件下的吸收利用机制及氮代谢响应研究提供依据。
关键词:  多形微眼藻  氮代谢  转录组  褐潮
DOI:10.11693/hyhz20190400073
分类号:Q178
基金项目:国家重点研发计划,2017YFC1404402号;国家自然科学基金,41976133号;青岛海洋科学与技术国家实验室鳌山卫科技创新计划项目,2016ASKJ02号。
NITROGEN METABOLISM PATHWAY OF MINUTOCELLUS POLYMORPHUS UNDER TWO NITROGEN NUTRITION CONDITIONS BASED ON TRANSCRIPTOME
LI Ying1,2, MI Tie-Zhu1,3,2, QIAO Ling4, ZHEN Yu1,3,2
1.Key Laboratory of Marine Environment and Ecology, Ministry of Education, Qingdao 266100, China;2.College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China;3.Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266071, China;4.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
Abstract:
To understand molecular mechanism of different nitrogen sources utilization in nitrogen metabolism pathway of Minutocellus polymorphus, we analyzed the transcriptome under the conditions of two nitrogen source nutrients (nitrate and urea) in Illumina-Hiseq2000 platform sequencing technology and studied the regulation of nitrogen metabolism in M. polymorphus by Gene Ontology (GO) annotation and Digital Gene Expression Profiling (DGE). We constructed the nitrogen metabolism pathway of M. polymorphus involving 15 enzymes and 76 coding genes. The results show that 10 enzyme-encoding genes were differentially expressed in the nitrogen metabolism pathway under different nitrogen conditions. The most significant differentially expressed were glutamate synthase (GLT1, GLTD), glutamate dehydrogenase (GDH), and glutamine synthetase (GS) related genes. The differential expressions of nitrate reductase and nitrite reductase in the urea culture medium was significantly higher than those in the nitrate culture solution, indicating that urea may have an impact on nitrate assimilation in the cells of M. polymorphus. These results provide a basis for studying the mechanism of nitrogen utilization and nitrogen metabolism response of diatom using different nitrogen species.
Key words:  Minutocellus polymorphus  nitrogen metabolism  RNA-seq  brown tide
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