孙敏红;卢晓鹏;曹雄军;李静;熊江;谢深喜;

• 1:湖南农业大学园艺园林学院
• 2:中南林业科技大学林学院

摘要(Abstract)：

【目的】进一步探讨枳橙对氮素吸收利用机制,为柑橘生产上提高氮素利用效率和科学施肥提供科学依据。【方法】以枳橙为材料,采用水培方法,在Hoagland配方的基础上进行调节,研究了不同形态氮素配比对枳橙幼苗硝酸还原酶(NR)活性变化及相关基因表达量的影响。【结果】幼苗叶片及根系NR活性均随着培养时间的延长而增加,且叶片NR活性高于根系。其中2号(NO_3~-∶NH_4~+=7∶3)和3号(NO_3~-∶NH_4~+=5∶5)处理的叶片酶活最高;1号(NO_3~-∶NH_4~+=10∶0)和2号(NO_3~-∶NH_4~+=7∶3)处理根系的NR活性显著高于其他处理;进一步研究NR相关基因表达情况表明:3个基因(CitNia1、CitNia2、CitNia OA)在不同处理的枳橙幼苗体内均有表达,其中CitNia1和CitNia2分别是根系和叶片NR相关主效基因,2者均受到NO_3~-、NH_4~+配比诱导,且3号处理(NO_3~-∶NH_4~+=5∶5)下表达量最高。【结论】枳橙幼苗硝态氮代谢的主要部位为叶片,且CitNia2是主效基因,通过调节铵硝比例可以促进其表达。

关键词(KeyWords)： 枳橙;硝态氮;铵态氮;硝酸还原酶

Abstract:

Keywords:

基金项目(Foundation): 现代农业(柑橘)产业技术体系(CARS-027);; 国家自然科学基金(31401824);; 教育部高校博士点基金(20134320120006)

作者(Author): 孙敏红;卢晓鹏;曹雄军;李静;熊江;谢深喜;

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参考文献(References)：

• [1]王川.中国柑橘生产与消费现状分析[J].农业展望,2006,2(1):8-12.WANG Chuan.Analysis of Citrus production and consumption in China[J].Agricultural Outlook,2006,2(1):8-12.
• [2]廖炜,李先信,阳志慧,卜汉文.氮磷肥对柑橘的影响研究进展[J].湖南农业科学,2010,24(12):34-35.LIAO Wei,LI Xianxin,YANG Zhihui,BU Hanwen.The advanced on nitrogen and phosphate on Citrus effect[J].Hunan Agricultural Science,2010,24(12):34-35.
• [3]AMTMANN A,ARMENGAUD P.Effects of N,P,K and S on metabolism:new knowledge gained from multi-level analysis[J].Current Opinion in Plant Biology,2009,12(3):275-283.
• [4]阳剑,时亚文,李宙炜,陶优生,唐启源.水稻碳氮代谢研究进展[J].作物研究,2011,25(4):383-387.YANG Jian,SHI Yawen,LI Zhouwei,TAO Yousheng,TANG Qiyuan.Advances in carbon and nitrogen metabolism in rice[J].Crop Research,2011,25(4):383-387.
• [5]王小纯,熊淑萍,马新明,张娟娟,王志强.不同形态氮素对专用型小麦花后氮代谢关键酶活性及籽粒蛋白质含量的影响[J].生态学报,2005,25(4):802-807.WANG Xiaochun,XIONG Shuping,MA Xinming,ZHANG Juanjuan,WANG Zhiqiang.Effects of different nitrogen forms on key enzyme activity involved in nitrogen metabolism and grain protein content in speciality wheat cultivars[J].Acta Ecological Sinica,2005,25(4):802-807.
• [6]DALLING M J,BOLAND G,WILLSON J H.Relation between acid proteinase activity and redistribution of nitrogen during grain development in wheat[J].Functional Plant Biology,1976,3(6):721-730.
• [7]张瑞珍,张恩和,孙长占.不同基因型玉米品种氮素营养效率差异的研究[J].吉林农业大学学报,2003,25(2):183-186.ZHANG Ruizhen,ZHANG Enhe,SUN Changzhan.Study different genotype maize variety on diversity of nitrogen nutrition efficience[J].Journal of Jilin Agricultural University,2003,25(2):183-186.
• [8]KUBO Y,OGURA N,NAKAGAWA H.Limited proteolysis of the nitratere ductase from spinach leaves[J].Journal of Biological Chemistry,1998,263(36):19684-19689.
• [9]VAUCHERET H,VINCENTZ M,KRONENBERGER J,ROUZE P.Molecular cloning and characterization of the two homologous genes coding for nitrate reductase in tobacco[J].Molecular&General Genetics Mgg,1989,216(1):10-15.
• [10]CHENG C L,DEWDNEY J,NAM H G,BGDEN B,GOODMAN H M.A new locus(NIAl)in Arabidopsis thaliana encoding nitrate reductase[J].Embo Journal,1988,7(11):3309-3314.
• [11]丁广洲,侯静,陈丽,马凤鸣,陈连江.甜菜nia基因的克隆及不同氮素形态诱导的差异表达[J].作物学报,2011,37(11):1949-1955.DING Guangzhou,HOU Jing,CHEN Li,MA Fengming,CHEN Lianjiang.Cloning of nia gene and its differential expression induced by different nitrogen forms in sugar beet(Beta vulgaris L.)[J].Acta Agronomica Sinica,2011,37(11):1949-1955.
• [12]XU Q,CHEN L L,RUAN X A,ZHU A.The draft genome of sweet orange(Citrus sinensis)[J].Nature Genetics,2013,45(1):59-66.
• [13]李合生.植物生理生化实验原理和技术[M].北京:高等教育出版社,2000:125-127.LI Hesheng.Principles and techniques of plant physiology and biochemistry experiment[M].Beijing:Higher Education Press,2000:125-127.
• [14]陈煜,朱保葛,张敬.不同氮源对大豆硝酸还原酶和谷氨酰胺合成酶活性及蛋白质含量的影响[J].大豆科学,2004,23(2):143-146.CHEN Yu,ZHU Baoge,ZHANG Jing.Effects of different nitrogens on activities of nitrate reductase,glutamine synthetase and seed protein vontents in soybean vultivars[J].Soybean Science,2004,23(2):143-146.
• [15]肖凯,张树华,邹定辉,张荣铣.不同形态氮素对小麦光合特性的影响[J].作物学报,2000,26(1):53-58.XIAO Kai,ZHANG Shuhua,ZOU Dinghui,ZHANG Rongxian.The effects of different nitrogen nutrition forms on photosynthetic characteristics in wheat leaves[J].Acta Agronomica Sinica,2000,26(1):53-58.
• [16]汪建飞,董彩霞,谢越,沈其荣.铵硝比和磷素营养对菠菜生长、氮素吸收和相关酶活性的影响[J].土壤学报,2006,46(6):954-955.WANG Jianfei,DONG Caixia,XIE Yue,SHEN Qirong.Effects of NH+4-N/NO-3-N ratio and phosphorus levels on growth,nitrogen uptake and relevant enzymes activity of spinach(Spinacia oleracea L.)[J].Acta Pedologica Sinica,2006,46(6):954-955.
• [17]张新要,刘卫群,易建华,李天福,陈良存,周文辉.红壤、水稻土上不同氮素形态配比对烤烟碳氮代谢关键酶活性的影响[J].云南农业大学学报,2005,20(2):225-230.ZHANG Xinyao,LIU Weiqun,YI Jianhua,LI Tianfu,CHEN Liangcun,ZHOU Wenhui.Effects of different forms of nitrogen fertilizer on predominant enzymes of C and N metabolism in red and rice soils of flue-cured tobacco[J].Journal of Yunnan Agricultural University,2005,20(2):225-230.
• [18]GASHAW L,MUGWIRA L M.Ammonium-N and nitrate-N effects on the growth and mineral compositions of tritical wheat and rye[J].Agronomy Journal,1981,73(1):47-51.
• [19]胡润芳,张广庆,滕振勇,林国强.不同形态氮素对大豆硝酸还原酶和谷氨酰胺合成酶活性及蛋白质含量的影响[J].东北农业大学学报,2012,43(1):31-32.HU Runfang,ZHANG Guangqing,TENG Zhenyong,LIN Guoqiang.Effect of different nitrogens on activities of nitrate reductase,glutamine synthetase,and seed protein contents in soybean cultivars[J].Journal of Northeast Agricultural University,2012,43(1):31-32.
• [20]李宪利,高东升,米瑞芙.苹果植株硝酸还原酶(NR)研究初报[J].山东农业大学学报,1997,28(1):1-4.LI Xianli,GAO Dongsheng,MI Ruifu.Study of the nitrate reductase activity of apple trees in orchard[J].Journal of Shandong Agricultural University,1997,28(1):1-4.
• [21]戴廷波,曹卫星,孙传范,姜东,荆奇.增铵营养对小麦光合作用及硝酸还原酶和谷氨酰胺合成酶的影响[J].应用生态学报,2003,14(9):1529-1532.DAI Tingbo,CAO Weixing,SUN Chuanfan,JIANG Dong,JING Qi.Effect of enhanced ammonium nutrition on photosynthesis and nitrate reduetase and glutamine synthetase activities of winter wheat[J].Chinese Journal of Applied Ecology,2003,14(9):1529-1532.
• [22]KAFKAFI U.Root temperature,concentration and the ratio NO3-/NH4+effect on plant development[J].Journal of Plant Nutrition,1990,13(10):1291-1306.
• [23]MUNZAROVA E,LORENZENB B,BRIXB H,VOJTISKOVA L,VOTRUBOVA O.Effect of NH4+/NO3-availability on nitrate reductase activity and nitrogen accumulation in wetland helophytes Phragmites australis and Glyceria maxima[J].Environmental&Experimental Botany,2006,55(1/2):49-60.
• [24]VINCENTZ M,MOUREAUX T,VAUCHERET H,CABOCHE M.Regulation of nitrate and nitrite reductase expression in Nicotiana plumbaginifolia leaves by nitrogen and carbon metabolites[J].Plant Journal,1993,3(2):315-324.
• [25]LOQUE D,TILLARD P,GOJON A,LEPETI M.Gene expression of the NO3-transporter NRT1.1 and the nitrate reductase NIA1 is repressed in Arabidopsis roots by NO2-,the product of NO3-reduction[J].Plant Physiology,2003,132(2):958-967.
• [26]ZOU D H.Effects of elevated atmospheric CO2on growth,photosynthesis and nitrogen metabolism in the economic brown seaweed,Hizikia fusiforme(Sargassaceae,Phaeophyta)[J].Aquaculture,2005,250(3/4):726-735.
• [27]WANG R,GUEGLER K,LABRIE S T,CRAWFORD N M.Genomic analysis of a nutrient response in Arabidopsis reveals diverse expression patterns and novel metabolic and potential regulatory genes induced by nitrate[J].Plant Cell,2000,12(8):1491-1509.
• [28]SIVASANKAR S,ROTHSTEIN S,OAKS A.Regulation of the accumulation and reduction of nitrate by nitrogen and carbon metabolites in maize seedlings[J].Plant Physiology,1997,114(2):583-589.
• [29]CHEN B M,WANG Z H,LI S X,WANG G X.Effect of nitrate supply on plant growth,nitrate accumulation,metabolic nitrate concentration and nitrate reductase activity in three leafy vegetables[J].Plant Science,2004,167(3):635-643.
• [30]曹云,范晓荣,孙淑斌,徐国华,沈其荣,狄廷君.增硝营养对不同基因型水稻苗期硝酸还原酶活性及其表达量的影响[J].植物营养与肥料学报,2007,13(1):99-105.CAO Yun,FAN Xiaorong,SUN Shubin,XU Guohua,SHEN Qirong,DI Tingjun.Effect of partial replacement of NH4+by NO3-on nitrate reductase activity and their genetic expression patterns in rice[J].Plant Nutrition and Fertilizer Science,2007,13(1):99-105.
• [31]LOPPES R,RADOUX M,OHRESSER M C P,MATAGNE R F.Transcriptional regulation of the NIA1 gene encoding nitrate reductase in Chlamydomonas reinhardtii:effects of various environmental factors on the expression of a reporter gene under the control of the Nia1 promoter[J].Plant Molecular Biology,1999,41(5):701-711.
• [32]WILKINSON J Q,CRAWFORD N M.Identification and characterization of a chlorate resistant mutant of Arabidopsis with mutations in both Nia1 and Nia2 nitrate reductase structural genes[J].Molecular&General Genetics Mgg,1993,239(1/2):289-297.