种植行距和品种对玉米根际反硝化菌群丰度和功能的影响
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摘要
反硝化是根际氮素损失重要途径,作物品种和行距改变是否会对根际反硝化产生影响尚不清楚。本研究比较了不同玉米品种和种植行距间根际土壤反硝化菌群丰度和功能的差异,为降低根际反硝化损失和提高氮肥利用效率提供科学依据。通过两个独立的田间试验,利用生物化学和分子微生物学方法,分别研究‘浚单20’、‘安农8号’、‘郑单958’、‘品玉18’和‘隆平206’5个玉米品种以及20 cm、30 cm、40 cm、50 cm种植行距对根际土壤反硝化能力、反硝化菌群丰度、N_2O/(N_2O+N_2)产物比和土壤呼吸等指标的影响。‘浚单20’、‘安农8号’、‘郑单958’根际反硝化能力显著低于其他两个品种;随着行距减少,反硝化能力有显著增加趋势。‘隆平206’和‘品玉18’的nir S型反硝化菌群丰度显著高于其他品种,而nir K和nosZ型菌群的丰度以‘浚单20’和‘安农8号’最高;行距20 cm的nir S和nir K型菌丰度显著高于其他行距处理,但nosZ型菌丰度以40 cm行距丰度最大。品种对N_2O/(N_2O+N_2)产物比有一定影响,其中‘安农8号’最低,但行距对产物比没有显著影响。相关分析表明反硝化能力与土壤呼吸和nirS型菌群丰度均极显著正相关,但与nos Z和nir K型菌群未呈现这种关系,由此表明nirS型菌丰度和根际有机碳差异可能是造成反硝化能力不同的主导因子。品种和种植行距会对玉米根际反硝化过程产生一定影响,根际低反硝化损失品种的筛选、选育和根际反硝化过程调控是减少根际反硝化损失,提高氮肥利用效率的有效途径。
Denitrification is a key pathway of nitrogen loss in the rhizosphere. It is unknown whether changes in variety and row spacing of crop have any impacts on denitrification in the rhizosphere. Thus this study compared the differences in abundance and function of denitrifiers in the rhizosphere of different maize varieties and row spacings. The aim of the study was to provide scientific basis for reducing rhizosphere denitrification losses and improving nitrogen use efficiency. Based on data from two independent field experiments, the study investigated the effects of five maize varieties(‘Xundan 20', ‘Annong 8', ‘Zhengdan 958', ‘Pinyu 18' and ‘Longping 206') and four row spacings(20 cm, 30 cm, 40 cm and 50 cm) on denitrification capacity, abundance of denitrifiers, N_2O/(N_2O+N_2) ratio and soil respiration in the rhizosphere using biochemical and molecular microbiological methods. Denitrification capacities in the rhizospheres of ‘Xundan 20', ‘Annong 8' and ‘Zhengdan 958' were significantly lower than those in the rhizosphere of the other maize varieties. With decreasing row spacing, denitrification capacity in rhizosphere soil obviously increased. The abundances of nir S-denitrifiers in the rhizospheres of ‘Longping 206' and ‘Pinyu 18' were significantly higher than those in the rhizospheres of the other maize varieties. The abundances of nir K- and nos Z-denitrifiers in the rhizospheres of ‘Xundan 20' and ‘Annong 8' were the highest. In all the row spacings, 20 cm had the highest abundances of nir S- and nir K-denitrifiers. As for nos Z-denitrifiers, the most abundance was found in the 40 cm row spacing treatment. The change of variety had a certain impact on N_2O/(N_2O+N_2) ratios(e.g. ‘Annong 8' had the lowest ratio), but the effect of row spacing was no significant. Correlation analysis showed there was a significantly positive correlation between denitrification capacity and soil respiration or the abundance of nir S-denitrifiers. But not significant correlation was found between denitrification capacity and the abundances of nos Z or nir K-denitrifiers. This suggested that differences in the abundances of nir S-denitrifiers and organic carbon in the rhizosphere were the dominant factors causing the changes in denitrification capacity. Changes in variety and row spacing could affect denitrification of maize rhizosphere as well. The screening of varieties with low denitrification loss and the regulation of denitrification processes in the rhizosphere were effective ways of reducing rhizosphere denitrification loss and improving crop nitrogen use efficiency.
Denitrification is a key pathway of nitrogen loss in the rhizosphere. It is unknown whether changes in variety and row spacing of crop have any impacts on denitrification in the rhizosphere. Thus this study compared the differences in abundance and function of denitrifiers in the rhizosphere of different maize varieties and row spacings. The aim of the study was to provide scientific basis for reducing rhizosphere denitrification losses and improving nitrogen use efficiency. Based on data from two independent field experiments, the study investigated the effects of five maize varieties(‘Xundan 20', ‘Annong 8', ‘Zhengdan 958', ‘Pinyu 18' and ‘Longping 206') and four row spacings(20 cm, 30 cm, 40 cm and 50 cm) on denitrification capacity, abundance of denitrifiers, N_2O/(N_2O+N_2) ratio and soil respiration in the rhizosphere using biochemical and molecular microbiological methods. Denitrification capacities in the rhizospheres of ‘Xundan 20', ‘Annong 8' and ‘Zhengdan 958' were significantly lower than those in the rhizosphere of the other maize varieties. With decreasing row spacing, denitrification capacity in rhizosphere soil obviously increased. The abundances of nir S-denitrifiers in the rhizospheres of ‘Longping 206' and ‘Pinyu 18' were significantly higher than those in the rhizospheres of the other maize varieties. The abundances of nir K- and nos Z-denitrifiers in the rhizospheres of ‘Xundan 20' and ‘Annong 8' were the highest. In all the row spacings, 20 cm had the highest abundances of nir S- and nir K-denitrifiers. As for nos Z-denitrifiers, the most abundance was found in the 40 cm row spacing treatment. The change of variety had a certain impact on N_2O/(N_2O+N_2) ratios(e.g. ‘Annong 8' had the lowest ratio), but the effect of row spacing was no significant. Correlation analysis showed there was a significantly positive correlation between denitrification capacity and soil respiration or the abundance of nir S-denitrifiers. But not significant correlation was found between denitrification capacity and the abundances of nos Z or nir K-denitrifiers. This suggested that differences in the abundances of nir S-denitrifiers and organic carbon in the rhizosphere were the dominant factors causing the changes in denitrification capacity. Changes in variety and row spacing could affect denitrification of maize rhizosphere as well. The screening of varieties with low denitrification loss and the regulation of denitrification processes in the rhizosphere were effective ways of reducing rhizosphere denitrification loss and improving crop nitrogen use efficiency.
引文
[1]陆雅海,张福锁.根际微生物研究进展[J].土壤,2006,38(2):113–121Lu Y H,Zhang F S.The advances in rhizosphere microbiology[J].Soils,2006,38(2):113–121
[2]Bothe H,Ferguson S J,Newton W E.Biology of the Nitrogen Cycle[M].Amsterdam:Elsevier Science,2007:1–427
[3]Ravishankara A R,Daniel J S,Portmann R W.Nitrous oxide(N2O):The dominant ozone depleting substance emitted in the21st century[J].Science,2009,326(5949):123–125
[4]Philippot L,Hallin S,Schlote M.Ecology of denitrifying prokaryotes in agricultural soil[J].Advances in Agronomy,2007,96:249–305
[5]宋贺,王成雨,陈清,等.长期秸秆还田对设施菜田土壤反硝化特征和N2O排放的影响[J].中国农业气象,2014,35(6):628–634Song H,Wang C Y,Chen Q,et al.Effects of long-term amendment of residue on denitrification characteristics and N2O emissions in greenhouse soil[J].Chinese Journal of Agrometeorology,2014,35(6):628–634
[6]Henry S,Texier S,Hallet S,et al.Disentangling the rhizosphere effect on nitrate reducers and denitrifiers:Insight into the role of root exudates[J].Environmental Microbiology,2008,10(11):3082–3092
[7]Hamonts K,Clough T J,Stewart A,et al.Effect of nitrogen and waterlogging on denitrifier gene abundance,community structure and activity in the rhizosphere of wheat[J].FEMS Microbiology Ecology,2013,83(3):568–584
[8]Wang H T,Su J Q,Zheng T L,et al.Impacts of vegetation,tidal process,and depth on the activities,abundances,and community compositions of denitrifiers in mangrove sediment[J].Applied Microbiology and Biotechnology,2014,98(22):9375–9387
[9]盛婧,张鹏,孙国锋,等.基于污染控制的小麦品种氮磷钾吸收与移除特征研究[J].生态环境学报,2015,24(3):487–493Sheng J,Zhang P,Sun G F,et al.Nutrient absorption characteristics and removal from soil with different wheat varieties based on pollution control[J].Ecology and Environmental Sciences,2015,24(3):487–493
[10]Liu J G,Qian M,Cai G L,et al.Variations between rice cultivars in root secretion of organic acids and the relationship with plant cadmium uptake[J].Environmental Geochemistry and Health,2007,29(3):189–195
[11]Li X G,Zhang T L,Wang X X,et al.The composition of root exudates from two different resistant peanut cultivars and their effects on the growth of soil-borne pathogen[J].International Journal of Biological Sciences,2013,9(2):164–173
[12]姜兴芳,陶洪斌,郑志芳,等.株行距配置对玉米根系性状及产量的影响[J].玉米科学,2013,21(2):116–121Jiang X F,Tao H B,Zheng Z F,et al.Effect of spacing allocation on the root system characters and yield of maize[J].Journal of Maize Sciences,2013,21(2):116–121
[13]Jiang W S,Wang K J,Wu Q P,et al.Effects of narrow plant spacing on root distribution and physiological nitrogen use efficiency in summer maize[J].The Crop Journal,2013,1(1):77–83
[14]王成雨,宋贺,胡玲惠,等.玉米品种耐淹形态指标筛选及其耐淹光合生理特性研究[J].安徽农业大学学报,2014,41(4):533–539Wang C Y,Song H,Hu L H,et al.Screening for cultivars with water-logging resistance through analyzing morphological indexes and photosynthetic characteristics in maize[J].Journal of Anhui Agricultural University,2014,41(4):533–539
[15]云鹏,高翔,陈磊,等.冬小麦–夏玉米轮作体系中不同施氮水平对玉米生长及其根际土壤氮的影响[J].植物营养与肥料学报,2010,16(3):567–574Yun P,Gao X,Chen L,et al.Plant nitrogen utilization and soil nitrogen status in rhizosphere of maize as affected by various nitrogen rates in wheat-maize rotation system[J].Plant Nutrition and Fertilizer Science,2010,16(3):567–574
[16]Braker G,Fesefeldt A,Witzel K P.Development of PCR primer systems for amplification of nitrite reductase genes(nir K and nir S)to detect denitrifying bacteria in environmental samples[J].Applied and Environmental Microbiology,1998,64(10):3769–3775
[17]Throb?ck I N,Enwall K,Jarvis A,et al.Reassessing PCR primers targeting nir S,nir K and nos Z genes for community surveys of denitrifying bacteria with DGGE[J].FEMS Microbiology Ecology,2004,49(3):401–417
[18]Henry S,Bru D,Stres B,et al.Quantitative detection of the nos Z gene,encoding nitrous oxide reductase,and comparison of the abundance of 16S r RNA,nar G,nir K,and nos Z genes in soils[J].Applied and Environmental Microbiology,2006,72(8):5181–5189
[19]王晓辉.生物炭对设施栽培土壤硝化和反硝化微生物群落的影响研究[D].北京:中国科学院大学,2013:27–29Wang X H.Effect of biochar on nitrifying and denitrifying communities in greenhouse soils[D].Beijing:University of Chinese Academy of Sciences,2013:27–29
[20]?imek M,Kal?ík J.Carbon and nitrate utilization in soils:The effect of long-term fertilization on potential denitrification[J].Geoderma,1998,83(3/4):269–280
[21]?imek M,Hopkins D W.Regulation of potential denitrification by soil p H in long-term fertilized arable soils[J].Biology and Fertility of Soils,1999,30(1/2):41–47
[22]Zhou X G,Guan S N,Wu F Z.Composition of soil microbial communities in the rhizosphere of cucumber cultivars with differing nitrogen acquisition efficiency[J].Applied Soil Ecology,2015,95:90–98
[23]Williams A,Davis A S,Ewing P M,et al.Precision control of soil nitrogen cycling via soil functional zone management[J].Agriculture,Ecosystems&Environment,2016,231:291–295
[24]Ohunakin A O,Adebisi S O.Nitrogen use and assimilate partitioning ability for grain yield in some selected maize(Zea mays L.)inbred lines genotypes[J].American Journal of Experimental Agriculture,2015,5(2):148–155
[25]Monneveux P,Zaidi P H,Sanchez C.Population density and low nitrogen affects yield-associated traits in tropical maize[J].Crop Science,2005,45(2):535–545
[26]Al-Naggar A M M,Shabana R A,Atta M M M,et al.Maize response to elevated plant density combined with lowered N-fertilizer rate is genotype-dependent[J].The Crop Journal,2015,3(2):96–109
[27]Boomsma C R,Santini J B,Tollenaar M,et al.Maize morphophysiological responses to intense crowding and low nitrogen availability:An analysis and review[J].Agronomy Journal,2009,101(6):1426–1452
[28]Wang W J,Dalal R C,Moody P W,et al.Relationships of soil respiration to microbial biomass,substrate availability and clay content[J].Soil Biology Biochemistry,2003,35(2):273–284
[29]Zeng F R,Chen S,Miao Y,et al.Changes of organic acid exudation and rhizosphere p H in rice plants under chromium stress[J].Environmental Pollution,2008,155(2):284–289
[30]Raut N,D?rsch P,Sitaula B K,et al.Soil acidification by intensified crop production in south Asia results in higher N2O/(N2O+N2)product ratios of denitrification[J].Soil Biology and Biochemistry,2012,55:104–112
[31]Yoshida M,Ishii S,Otsuka S,et al.nir K-harboring denitrifiers are more responsive to denitrification-inducing conditions in rice paddy soil than nir S-harboring bacteria[J].Microbes and Environments,2010,25(1):45–48
[32]Enwall K,Throb?ck I N,Stenberg M,et al.Soil resources influence spatial patterns of denitrifying communities at scales compatible with land management[J].Applied and Environmental Microbiology,2010,76(7):2243–2250
[33]Ducey T F,Shriner A D,Hunt P G.Nitrification and denitrification gene abundances in swine wastewater anaerobic lagoons[J].Journal of Environmental Quality,2011,40(2):610–619
[34]Liu B B,M?rkved P T,Frosteg?rd?,et al.Denitrification gene pools,transcription and kinetics of NO,N2O and N production as affected by soil p H[J].FEMS Microbiology Ecology,2010,72(3):407–417
[35]Billings S A,Tiemann L K.Warming-induced enhancement of soil N2O efflux linked to distinct response times of genes driving N2O production and consumption[J].Biogeochemistry,2014,119(1/3):371–386
[2]Bothe H,Ferguson S J,Newton W E.Biology of the Nitrogen Cycle[M].Amsterdam:Elsevier Science,2007:1–427
[3]Ravishankara A R,Daniel J S,Portmann R W.Nitrous oxide(N2O):The dominant ozone depleting substance emitted in the21st century[J].Science,2009,326(5949):123–125
[4]Philippot L,Hallin S,Schlote M.Ecology of denitrifying prokaryotes in agricultural soil[J].Advances in Agronomy,2007,96:249–305
[5]宋贺,王成雨,陈清,等.长期秸秆还田对设施菜田土壤反硝化特征和N2O排放的影响[J].中国农业气象,2014,35(6):628–634Song H,Wang C Y,Chen Q,et al.Effects of long-term amendment of residue on denitrification characteristics and N2O emissions in greenhouse soil[J].Chinese Journal of Agrometeorology,2014,35(6):628–634
[6]Henry S,Texier S,Hallet S,et al.Disentangling the rhizosphere effect on nitrate reducers and denitrifiers:Insight into the role of root exudates[J].Environmental Microbiology,2008,10(11):3082–3092
[7]Hamonts K,Clough T J,Stewart A,et al.Effect of nitrogen and waterlogging on denitrifier gene abundance,community structure and activity in the rhizosphere of wheat[J].FEMS Microbiology Ecology,2013,83(3):568–584
[8]Wang H T,Su J Q,Zheng T L,et al.Impacts of vegetation,tidal process,and depth on the activities,abundances,and community compositions of denitrifiers in mangrove sediment[J].Applied Microbiology and Biotechnology,2014,98(22):9375–9387
[9]盛婧,张鹏,孙国锋,等.基于污染控制的小麦品种氮磷钾吸收与移除特征研究[J].生态环境学报,2015,24(3):487–493Sheng J,Zhang P,Sun G F,et al.Nutrient absorption characteristics and removal from soil with different wheat varieties based on pollution control[J].Ecology and Environmental Sciences,2015,24(3):487–493
[10]Liu J G,Qian M,Cai G L,et al.Variations between rice cultivars in root secretion of organic acids and the relationship with plant cadmium uptake[J].Environmental Geochemistry and Health,2007,29(3):189–195
[11]Li X G,Zhang T L,Wang X X,et al.The composition of root exudates from two different resistant peanut cultivars and their effects on the growth of soil-borne pathogen[J].International Journal of Biological Sciences,2013,9(2):164–173
[12]姜兴芳,陶洪斌,郑志芳,等.株行距配置对玉米根系性状及产量的影响[J].玉米科学,2013,21(2):116–121Jiang X F,Tao H B,Zheng Z F,et al.Effect of spacing allocation on the root system characters and yield of maize[J].Journal of Maize Sciences,2013,21(2):116–121
[13]Jiang W S,Wang K J,Wu Q P,et al.Effects of narrow plant spacing on root distribution and physiological nitrogen use efficiency in summer maize[J].The Crop Journal,2013,1(1):77–83
[14]王成雨,宋贺,胡玲惠,等.玉米品种耐淹形态指标筛选及其耐淹光合生理特性研究[J].安徽农业大学学报,2014,41(4):533–539Wang C Y,Song H,Hu L H,et al.Screening for cultivars with water-logging resistance through analyzing morphological indexes and photosynthetic characteristics in maize[J].Journal of Anhui Agricultural University,2014,41(4):533–539
[15]云鹏,高翔,陈磊,等.冬小麦–夏玉米轮作体系中不同施氮水平对玉米生长及其根际土壤氮的影响[J].植物营养与肥料学报,2010,16(3):567–574Yun P,Gao X,Chen L,et al.Plant nitrogen utilization and soil nitrogen status in rhizosphere of maize as affected by various nitrogen rates in wheat-maize rotation system[J].Plant Nutrition and Fertilizer Science,2010,16(3):567–574
[16]Braker G,Fesefeldt A,Witzel K P.Development of PCR primer systems for amplification of nitrite reductase genes(nir K and nir S)to detect denitrifying bacteria in environmental samples[J].Applied and Environmental Microbiology,1998,64(10):3769–3775
[17]Throb?ck I N,Enwall K,Jarvis A,et al.Reassessing PCR primers targeting nir S,nir K and nos Z genes for community surveys of denitrifying bacteria with DGGE[J].FEMS Microbiology Ecology,2004,49(3):401–417
[18]Henry S,Bru D,Stres B,et al.Quantitative detection of the nos Z gene,encoding nitrous oxide reductase,and comparison of the abundance of 16S r RNA,nar G,nir K,and nos Z genes in soils[J].Applied and Environmental Microbiology,2006,72(8):5181–5189
[19]王晓辉.生物炭对设施栽培土壤硝化和反硝化微生物群落的影响研究[D].北京:中国科学院大学,2013:27–29Wang X H.Effect of biochar on nitrifying and denitrifying communities in greenhouse soils[D].Beijing:University of Chinese Academy of Sciences,2013:27–29
[20]?imek M,Kal?ík J.Carbon and nitrate utilization in soils:The effect of long-term fertilization on potential denitrification[J].Geoderma,1998,83(3/4):269–280
[21]?imek M,Hopkins D W.Regulation of potential denitrification by soil p H in long-term fertilized arable soils[J].Biology and Fertility of Soils,1999,30(1/2):41–47
[22]Zhou X G,Guan S N,Wu F Z.Composition of soil microbial communities in the rhizosphere of cucumber cultivars with differing nitrogen acquisition efficiency[J].Applied Soil Ecology,2015,95:90–98
[23]Williams A,Davis A S,Ewing P M,et al.Precision control of soil nitrogen cycling via soil functional zone management[J].Agriculture,Ecosystems&Environment,2016,231:291–295
[24]Ohunakin A O,Adebisi S O.Nitrogen use and assimilate partitioning ability for grain yield in some selected maize(Zea mays L.)inbred lines genotypes[J].American Journal of Experimental Agriculture,2015,5(2):148–155
[25]Monneveux P,Zaidi P H,Sanchez C.Population density and low nitrogen affects yield-associated traits in tropical maize[J].Crop Science,2005,45(2):535–545
[26]Al-Naggar A M M,Shabana R A,Atta M M M,et al.Maize response to elevated plant density combined with lowered N-fertilizer rate is genotype-dependent[J].The Crop Journal,2015,3(2):96–109
[27]Boomsma C R,Santini J B,Tollenaar M,et al.Maize morphophysiological responses to intense crowding and low nitrogen availability:An analysis and review[J].Agronomy Journal,2009,101(6):1426–1452
[28]Wang W J,Dalal R C,Moody P W,et al.Relationships of soil respiration to microbial biomass,substrate availability and clay content[J].Soil Biology Biochemistry,2003,35(2):273–284
[29]Zeng F R,Chen S,Miao Y,et al.Changes of organic acid exudation and rhizosphere p H in rice plants under chromium stress[J].Environmental Pollution,2008,155(2):284–289
[30]Raut N,D?rsch P,Sitaula B K,et al.Soil acidification by intensified crop production in south Asia results in higher N2O/(N2O+N2)product ratios of denitrification[J].Soil Biology and Biochemistry,2012,55:104–112
[31]Yoshida M,Ishii S,Otsuka S,et al.nir K-harboring denitrifiers are more responsive to denitrification-inducing conditions in rice paddy soil than nir S-harboring bacteria[J].Microbes and Environments,2010,25(1):45–48
[32]Enwall K,Throb?ck I N,Stenberg M,et al.Soil resources influence spatial patterns of denitrifying communities at scales compatible with land management[J].Applied and Environmental Microbiology,2010,76(7):2243–2250
[33]Ducey T F,Shriner A D,Hunt P G.Nitrification and denitrification gene abundances in swine wastewater anaerobic lagoons[J].Journal of Environmental Quality,2011,40(2):610–619
[34]Liu B B,M?rkved P T,Frosteg?rd?,et al.Denitrification gene pools,transcription and kinetics of NO,N2O and N production as affected by soil p H[J].FEMS Microbiology Ecology,2010,72(3):407–417
[35]Billings S A,Tiemann L K.Warming-induced enhancement of soil N2O efflux linked to distinct response times of genes driving N2O production and consumption[J].Biogeochemistry,2014,119(1/3):371–386