白肋烟烟叶氮同化能力及其对硝酸盐积累的影响
|
摘要
为明确白肋烟烟叶硝酸盐积累的主要原因,以烤烟为对照,研究白肋烟硝酸盐同化作用及其对硝酸盐积累的影响。结果表明:增施5倍氮素后,白肋烟较烤烟干物质积累量与烤烟相近;在相同供氮条件下,白肋烟烟叶硝酸盐含量较烤烟高103.36%,但氮素积累量,硝酸还原酶和谷氨酰胺合成酶活性,色素和总糖及还原糖含量,NH_4-N和可溶性蛋白质含量,基因GDHA_1、GS1、NIA1和NIA2表达水平均较烤烟低,氮还原同化能力弱。由相关性分析得出,烟叶硝酸盐含量与氮还原同化酶活性和两糖含量均呈现负相关,相关系数达极显著水平;可溶性蛋白质含量与氮还原同化酶活性和两糖含量均呈现正相关,相关系数达极显著水平。氮还原同化能力弱和能量物质供应不足是引起白肋烟硝酸盐积累的主要原因。
Experiment was set up to investigate nitrogen assimilation capacity of burley tobacco leaves and its effects on nitrate accumulation with ?ue-cured tobacco as contrast. Results showed that dry matter accumulation in burley tobacco was equivalent to that in?ue-cured tobacco when 5-fold amount of nitrogen was additionally applied. Nitrate content in burley tobacco was higher than that in ?uecured tobacco by 103.26 % at same nitrogen application level, however, nitrogen accumulation, nitrate reductase activity(NRA), glutamine synthetase activity(GSA), pigment content, total reducing soluble sugar and reducing soluble sugar content, NH_4-N and soluble protein content, genes(GDHA_1, GS1, NIA1, NIA2) expression level in burley tobacco were all signi?cantly lower than in ?ue-cured tobacco,indicating that nitrate assimilation capacity of burley tobacco was weaker than ?ue-cured tobacco. Correlation analysis revealed that nitrate content was signi?cantly and negatively correlated with NRA and total soluble sugar & reducing soluble sugar contents; while soluble protein content was in significant positive correlation with NRA, and total soluble sugar & reducing soluble sugar content. The weak nitrogen assimilation capacity and low energy supply are two major causes for nitrate accumulation in burley tobacco.
Experiment was set up to investigate nitrogen assimilation capacity of burley tobacco leaves and its effects on nitrate accumulation with ?ue-cured tobacco as contrast. Results showed that dry matter accumulation in burley tobacco was equivalent to that in?ue-cured tobacco when 5-fold amount of nitrogen was additionally applied. Nitrate content in burley tobacco was higher than that in ?uecured tobacco by 103.26 % at same nitrogen application level, however, nitrogen accumulation, nitrate reductase activity(NRA), glutamine synthetase activity(GSA), pigment content, total reducing soluble sugar and reducing soluble sugar content, NH_4-N and soluble protein content, genes(GDHA_1, GS1, NIA1, NIA2) expression level in burley tobacco were all signi?cantly lower than in ?ue-cured tobacco,indicating that nitrate assimilation capacity of burley tobacco was weaker than ?ue-cured tobacco. Correlation analysis revealed that nitrate content was signi?cantly and negatively correlated with NRA and total soluble sugar & reducing soluble sugar contents; while soluble protein content was in significant positive correlation with NRA, and total soluble sugar & reducing soluble sugar content. The weak nitrogen assimilation capacity and low energy supply are two major causes for nitrate accumulation in burley tobacco.
引文
[1]Chaplin JF.Inheritance and possible use of pale yellow character in tobacco[J].Crop Science,1969,9(2):169-172.
[2]景延秋.不同种植、调制条件下对白肋烟重要香味成分的影响[D].郑州:河南农业大学,2006.Jing Yanqiu.The Effects of different planting and air-curing conditions on important flavor components in burley tobacco leaves[D].Zhengzhou:Henan Agricultural University,2006.
[3]Li Y,Chang D,Sun J,et al.The analysis of the difference on nitrogen metabolism of flue-cured tobacco and burley seedling.Beijing,2016:244-244.
[4]尚志强.施氮量对白肋烟生长发育及产量质量的影响[J].中国农学通报,2007,23(1):299-301.Shang Zhiqiang.Effects of nitrogen amount on growth and development yield and quality in burley tobacco[J].Chinese Agricultural Science Bulletin,2007,23(1):299-301.
[5]Edwards KD,Fernandez-Pozo N,Drake-Stowe K,et al.A reference genome for Nicotiana tabacum enables map-based cloning of homeologous loci implicated in nitrogen utilization efficiency[J].Bmc Genomics,2017,18:14.
[6]Lewis RS,Parker RG,Danehower DA,et al.Impact of alleles at the yellow Burley(Yb)Loci and nitrogen fertilization rate on nitrogen utilization efficiency and tobacco-specific nitrosamine(TSNA)formation in air-cured tobacco[J].Journal of Agricultural and Food Chemistry,2012,60(25):6454-6461.
[7]Shi H,Wang R,Bush LP,et al.The relationships between TSNAs and their precursors in burley tobacco from different regions and varieties[J].Journal of Food,Agriculture&Environment,2012,10(3/4):1048-1052.
[8]Shi H,Wang R,Bush LP,et al.Changes in TSNA contents during tobacco storage and the effect of temperature and nitrate level on TSNA formation[J].Journal of Agricultural and Food Chemistry,2013,61(47):11588-11594.
[9]Fischer S,Spiegelhalder B,Preussmann R.Preformed tobaccospecific nitrosamines in tobacco--role of nitrate and influence of tobacco type[J].Carcinogenesis,1989,10(8):1511-7.
[10]Brauer EK,Rochon A,Bi Y,et al.Reappraisal of nitrogen use efficiency in rice overexpressing glutamine synthetase1[J].Physiologia Plantarum,2011,141(4):361-372.
[11]Zhang Z,Xiong S,Wei Y,et al.The role of glutamine synthetase isozymes in enhancing nitrogen use efficiency of N-efficient winter wheat[J].Scientific Reports,2017,7.
[12]Wang X,Wei Y,Shi L,et al.New isoforms and assembly of glutamine synthetase in the leaf of wheat(Triticum aestivum L.)[J].Journal of Experimental Botany,2015,66(21):6827-34.
[13]Miflin BJ,Habash DZ.The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops[J].Journal of Experimnetal Botany,2002,53:979-987.
[14]Balotf S,Kavoosi G,Kholdebarin B.Nitrate reductase,nitrite reductase,glutamine synthetase,and glutamate synthase expression and activity in response to different nitrogen sources in nitrogen-starved wheat seedlings[J].Biotechnol Appl Biochem,2016,63(2):220-229.
[15]Li Y,Yang H,Chang D,et al.Biochemical,physiological and transcriptomic comparison between burley and flue-cured tobacco seedlings in relation to carbohydrates and nitrate content[J].Molecules,2017,22(12):2126.
[16]O’Neal D,Joy K.Glutamine synthetase of pea leaves.I.Purification,stabilization,and pH optima.[J].Plant Physiology,1974,54(5):773-779.
[17]李合生.现代植物生理学[M].北京:高等教育出版社,2006.Li Hesheng.Principle and technology of plant physiological and biochemical experiments[M].Beijing:Higher Education Press,2006.
[18]范淑琴,梁淑文.现代植物生理学实验指南[M].上海:科学出版社,1999.Fang Shuqin,Liang Shuwen.Principle and technology of plant physiological and biochemical experiments:A Laboratory Manual[M].Shanghai:Science Press,1999.
[19]Cataldo DA,Haroon M,Schrader LE,et al.Rapid cplorimetric deternination of nitrate in plant-tissure by nitration of salicylic acid[J].Commun Soil Sci Plan,1975,6:71-80.
[20]史宏志,徐发华,杨兴有,等.不同产地和品种白肋烟烟草特有亚硝胺与前体物关系[J].中国烟草学报,2012,18(5):9-15.Shi Hongzhi,Xu Fahua,Yang Xingyou,et al.The relationships between TSNAs and their precursors in different burley tobacco varieties from different regions[J].Acta Tabacaria Sinica,2012,18(5):9-15.
[21]孙榅淑,王俊,周骏,等.硝态氮含量对烟叶高温贮藏过程TSNA形成的影响[J].中国烟草学报,2015,21(2):53-57.Sun Wenshu,Wang Jun,Zhou Jun,et al.Effect of nitrate nitrogen level in tobacco leaves on TSNAs formation during high temperature storage[J].Acta Tabacaria Sinica,2015,21(2):53-57.
[22]Li Y,Shi H,Yang H,et al.Difference between Burley Tobacco and Flue-Cured Tobacco in nitrate accumulation and chemical regulation of nitrate and tsna contents[J].Journal of Chemistry,2017,2017:13.
[23]Navarro FJ,Perdomo G,Tejera P,et al.The role of nitrate reductase in the regulation of the nitrate assimilation pathway in the yeast Hansenula polymorpha[J].Fems Yeast Research,2003,4(2):149-155.
[24]Man HM,Boriel R,EI-Khatib R,et al.Characterization of transgenic poplar with ectopic expression of pine cytosolic glutamine synthetase under conditions of varying nitrogen availability[J].New Phytol,2005,167:31-39.
[25]周建建,蔡晓锋,徐晨曦,等.不同菠菜基因型硝酸盐积累与氮吸收、利用效率的比较[J].上海师范大学学报(自然科学版),2017,5:611-617.Zhou Jianjian,Cai Xiaofeng,Xu Chenxi,et al.Comparison of nitrate accumulation,nitrogen uptake and utilization efficiency among different spinachn(Spinacia oleracea L.)genotypes[J].Journal of Shanghai Normal University(Natural Sciences),2017,5:611-617.
[26]Lu.J,Zhang L,Lewis RS,et al.Expression of a constitutively active nitrate reductase variant in tobacco reduces tobacco-specific nitrosamine accumulation in cured leaves and cigarette smoke[J].Plant Biotechnology Journal,2016,14(7):1500-1510.
[27]高相彬,赵凤霞,孟智勇,等.碳氮代谢与烟叶品质的关系及其氮素的响应研究进展[J].农学学报,2013,3(8):38-40.Gao Xiangbin,Zhao Fengxia,Meng Zhiyong,et al.A review on the response of relation between carbon and nitrogen metabolism and tobacco leaf quality to nitrogen[J].Journal of Agriculture,2013,3(8):38-40.
[28]史宏志,韩锦峰.烤烟碳氮代谢几个问题的探讨[J].烟草科技,1998,(2):34-36.Shi Hongzhi,Han Jinfeng.Discussion of several problem on carbon and nitrogen metabolism in Flue-cured tobacco[J].Tobacco science&technology,1998,(2):34-36.
[29]韩锦峰,史宏志,官春云,等.不同施氮水平和氮素来源烟叶碳氮比及其与碳氮代谢的关系[J].中国烟草学报,1996,3(1):19-25.Han Jinfeng,Shi Hongzhi,Guan Chunyun,et al.C/N of tobacco leaf and its relations with carbon and nitrogen metabolism as related to nitrogen level and source[J].1996,3(1):19-25.
[30]艾绍英,姚建武,黄小红,等.蔬菜硝酸盐的还原转化特性研究[J].植物营养与肥料学报,2002,8(1):40-43.Ai Shaoying,Yao Jianwu,Huang Xiaohong,et al.Study on the nitrate reduction characteristic of vegetables[J].Plant nutrition and fertilizer science.2002,8(1):40-43.
[2]景延秋.不同种植、调制条件下对白肋烟重要香味成分的影响[D].郑州:河南农业大学,2006.Jing Yanqiu.The Effects of different planting and air-curing conditions on important flavor components in burley tobacco leaves[D].Zhengzhou:Henan Agricultural University,2006.
[3]Li Y,Chang D,Sun J,et al.The analysis of the difference on nitrogen metabolism of flue-cured tobacco and burley seedling.Beijing,2016:244-244.
[4]尚志强.施氮量对白肋烟生长发育及产量质量的影响[J].中国农学通报,2007,23(1):299-301.Shang Zhiqiang.Effects of nitrogen amount on growth and development yield and quality in burley tobacco[J].Chinese Agricultural Science Bulletin,2007,23(1):299-301.
[5]Edwards KD,Fernandez-Pozo N,Drake-Stowe K,et al.A reference genome for Nicotiana tabacum enables map-based cloning of homeologous loci implicated in nitrogen utilization efficiency[J].Bmc Genomics,2017,18:14.
[6]Lewis RS,Parker RG,Danehower DA,et al.Impact of alleles at the yellow Burley(Yb)Loci and nitrogen fertilization rate on nitrogen utilization efficiency and tobacco-specific nitrosamine(TSNA)formation in air-cured tobacco[J].Journal of Agricultural and Food Chemistry,2012,60(25):6454-6461.
[7]Shi H,Wang R,Bush LP,et al.The relationships between TSNAs and their precursors in burley tobacco from different regions and varieties[J].Journal of Food,Agriculture&Environment,2012,10(3/4):1048-1052.
[8]Shi H,Wang R,Bush LP,et al.Changes in TSNA contents during tobacco storage and the effect of temperature and nitrate level on TSNA formation[J].Journal of Agricultural and Food Chemistry,2013,61(47):11588-11594.
[9]Fischer S,Spiegelhalder B,Preussmann R.Preformed tobaccospecific nitrosamines in tobacco--role of nitrate and influence of tobacco type[J].Carcinogenesis,1989,10(8):1511-7.
[10]Brauer EK,Rochon A,Bi Y,et al.Reappraisal of nitrogen use efficiency in rice overexpressing glutamine synthetase1[J].Physiologia Plantarum,2011,141(4):361-372.
[11]Zhang Z,Xiong S,Wei Y,et al.The role of glutamine synthetase isozymes in enhancing nitrogen use efficiency of N-efficient winter wheat[J].Scientific Reports,2017,7.
[12]Wang X,Wei Y,Shi L,et al.New isoforms and assembly of glutamine synthetase in the leaf of wheat(Triticum aestivum L.)[J].Journal of Experimental Botany,2015,66(21):6827-34.
[13]Miflin BJ,Habash DZ.The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops[J].Journal of Experimnetal Botany,2002,53:979-987.
[14]Balotf S,Kavoosi G,Kholdebarin B.Nitrate reductase,nitrite reductase,glutamine synthetase,and glutamate synthase expression and activity in response to different nitrogen sources in nitrogen-starved wheat seedlings[J].Biotechnol Appl Biochem,2016,63(2):220-229.
[15]Li Y,Yang H,Chang D,et al.Biochemical,physiological and transcriptomic comparison between burley and flue-cured tobacco seedlings in relation to carbohydrates and nitrate content[J].Molecules,2017,22(12):2126.
[16]O’Neal D,Joy K.Glutamine synthetase of pea leaves.I.Purification,stabilization,and pH optima.[J].Plant Physiology,1974,54(5):773-779.
[17]李合生.现代植物生理学[M].北京:高等教育出版社,2006.Li Hesheng.Principle and technology of plant physiological and biochemical experiments[M].Beijing:Higher Education Press,2006.
[18]范淑琴,梁淑文.现代植物生理学实验指南[M].上海:科学出版社,1999.Fang Shuqin,Liang Shuwen.Principle and technology of plant physiological and biochemical experiments:A Laboratory Manual[M].Shanghai:Science Press,1999.
[19]Cataldo DA,Haroon M,Schrader LE,et al.Rapid cplorimetric deternination of nitrate in plant-tissure by nitration of salicylic acid[J].Commun Soil Sci Plan,1975,6:71-80.
[20]史宏志,徐发华,杨兴有,等.不同产地和品种白肋烟烟草特有亚硝胺与前体物关系[J].中国烟草学报,2012,18(5):9-15.Shi Hongzhi,Xu Fahua,Yang Xingyou,et al.The relationships between TSNAs and their precursors in different burley tobacco varieties from different regions[J].Acta Tabacaria Sinica,2012,18(5):9-15.
[21]孙榅淑,王俊,周骏,等.硝态氮含量对烟叶高温贮藏过程TSNA形成的影响[J].中国烟草学报,2015,21(2):53-57.Sun Wenshu,Wang Jun,Zhou Jun,et al.Effect of nitrate nitrogen level in tobacco leaves on TSNAs formation during high temperature storage[J].Acta Tabacaria Sinica,2015,21(2):53-57.
[22]Li Y,Shi H,Yang H,et al.Difference between Burley Tobacco and Flue-Cured Tobacco in nitrate accumulation and chemical regulation of nitrate and tsna contents[J].Journal of Chemistry,2017,2017:13.
[23]Navarro FJ,Perdomo G,Tejera P,et al.The role of nitrate reductase in the regulation of the nitrate assimilation pathway in the yeast Hansenula polymorpha[J].Fems Yeast Research,2003,4(2):149-155.
[24]Man HM,Boriel R,EI-Khatib R,et al.Characterization of transgenic poplar with ectopic expression of pine cytosolic glutamine synthetase under conditions of varying nitrogen availability[J].New Phytol,2005,167:31-39.
[25]周建建,蔡晓锋,徐晨曦,等.不同菠菜基因型硝酸盐积累与氮吸收、利用效率的比较[J].上海师范大学学报(自然科学版),2017,5:611-617.Zhou Jianjian,Cai Xiaofeng,Xu Chenxi,et al.Comparison of nitrate accumulation,nitrogen uptake and utilization efficiency among different spinachn(Spinacia oleracea L.)genotypes[J].Journal of Shanghai Normal University(Natural Sciences),2017,5:611-617.
[26]Lu.J,Zhang L,Lewis RS,et al.Expression of a constitutively active nitrate reductase variant in tobacco reduces tobacco-specific nitrosamine accumulation in cured leaves and cigarette smoke[J].Plant Biotechnology Journal,2016,14(7):1500-1510.
[27]高相彬,赵凤霞,孟智勇,等.碳氮代谢与烟叶品质的关系及其氮素的响应研究进展[J].农学学报,2013,3(8):38-40.Gao Xiangbin,Zhao Fengxia,Meng Zhiyong,et al.A review on the response of relation between carbon and nitrogen metabolism and tobacco leaf quality to nitrogen[J].Journal of Agriculture,2013,3(8):38-40.
[28]史宏志,韩锦峰.烤烟碳氮代谢几个问题的探讨[J].烟草科技,1998,(2):34-36.Shi Hongzhi,Han Jinfeng.Discussion of several problem on carbon and nitrogen metabolism in Flue-cured tobacco[J].Tobacco science&technology,1998,(2):34-36.
[29]韩锦峰,史宏志,官春云,等.不同施氮水平和氮素来源烟叶碳氮比及其与碳氮代谢的关系[J].中国烟草学报,1996,3(1):19-25.Han Jinfeng,Shi Hongzhi,Guan Chunyun,et al.C/N of tobacco leaf and its relations with carbon and nitrogen metabolism as related to nitrogen level and source[J].1996,3(1):19-25.
[30]艾绍英,姚建武,黄小红,等.蔬菜硝酸盐的还原转化特性研究[J].植物营养与肥料学报,2002,8(1):40-43.Ai Shaoying,Yao Jianwu,Huang Xiaohong,et al.Study on the nitrate reduction characteristic of vegetables[J].Plant nutrition and fertilizer science.2002,8(1):40-43.