不同生物硝化抑制剂对红壤性水稻土N2O排放的影响及其机制

为揭示不同生物硝化抑制剂(BNIs)对红壤性水稻土N₂O排放的影响差异及作用机制,通过21 d的土柱淹水培养试验,比较了三种BNIs 1,9-癸二醇(1,9-D)、亚麻酸(LN)和3-(4-羟基苯基)丙酸甲酯(MHPP)与化学合成硝化抑制剂双氰胺(DCD)对土壤N₂O排放及相关硝化、反硝化功能基因的影响。结果表明:不同BNIs(1,9-D、LN、MHPP)可以显著平均降低土壤N₂O日排放峰值40.1%;1,9-D和MHPP可分别抑制N₂O排放总量44.5%和43.9%,而DCD和LN对N₂O排放总量没有显著影响。1,9-D和MHPP对AOA(氨氧化古菌)、AOB(氨氧化细菌)硝化菌和nirS、nirK型反硝化菌的调控均有所不同,1,9-D可以同时抑制AOA、AOB和nirS微生物的生长;MHPP仅可以抑制AOA的生长;其中,AOA-amoA和nirS基因丰度与土壤N₂O的排放呈显著正相关关系。同时,1,9-D和MHPP均增加了nosZ基因丰度及其与AOA-...     

水氮耦合对设施农田土壤自养和异养硝化作用的影响

水氮措施影响设施土壤氮素的转化及硝化微生物活性,但水氮耦合对设施土壤自养和异养硝化作用差异的影响尚不明确。以连续8年设施水氮耦合田间定位试验土壤为研究对象,控制不同土壤田间持水量(WHC)(40%WHC、60%WHC和80% WHC)进行室内微宇宙培养试验,通过添加乙炔抑制剂抑制自养硝化途径,研究水氮耦合对设施土壤自养和异养硝化速率及参与自养硝化的氨氧化微生物的影响,分析氨氧化微生物氨氧化古细菌(AOA)和氨氧化细菌(AOB)对自养硝化作用的贡献。结果表明,水氮耦合下,不同硝化途径NH4+-N、NO^3--N含量以及参与自养硝化的AOA amoA和AOB amoA基因拷贝数均有显著差异。无乙炔培养7 d后,NO^3--N含量显著增加,而NH4+-N含量显著降低,AOA amoA和AOB amoA的基因丰度显著增加。添加乙炔后,NO^3--N、NH4+-N含量基本保持恒定,AOA amoA和AOB amoA基因丰度显著减少。水氮耦合显著影响自养和异养硝化速率,冗余分析(RDA)表明,NH4+-N含量、AOB amoA、NO^3--N-C2H2、AOA amoA可分别解释自养和异养硝化速率变异的68.9%、34.9%、32.8%和24.4%。设施土壤存在自养硝化和异养硝化两种途径,60%～80%WHC各施氮处理均以自养硝化为主,占总硝化速率的65%～86%;仅40%WHC下,氮纯养分量300和525 kg·hm-2处理以异养硝化为主,占总硝化速率的61%～77%。AOB和AOA共同驱动自养硝化,且AOB贡献更大。     

紫色土硝化过程中氨氧化细菌和古菌对温度的响应

采用室内恒温培养法,探究4℃、15℃、28℃和40℃条件下中性紫色土硝化作用的差异及氨氧化细菌(AOB)和氨氧化古菌(AOA)对不同温度的响应模式。结果表明:28℃条件下硝化作用最强,净硝化率为3.40 mg N kg~(-1) d~(-1),40℃条件下硝化作用较弱,净硝化率为1.52 mg N kg~(-1) d~(-1),15℃条件下硝化作用发生延迟,净硝化率为2.40 mg N kg~(-1) d~(-1),4℃条件下硝化作用被抑制。28℃条件下的硝化动力学过程符合一级动力学模型(R2为0.996),15℃和40℃条件下的硝化动力学过程符合零级动力学模型(R2分别为0.969和0.984),说明15℃和40℃条件下限制硝化作用的因子可能是硝化微生物活性。AOB amoA基因拷贝数在28℃条件下达到最大,而AOA amoA基因拷贝数在40℃条件下达到最大。AOA和AOB amoA基因拷贝数在培养期间的变化表明AOB和AOA可能在中性紫色土硝化作用中共同发挥作用。在培养过程中,AOA与AOB amoA基因拷贝数之比在15℃和28℃条件下,由1 d时的7.38和8.92下降为28 d时的1.51和2.12,呈显著降低趋势(P 0.05);在40℃条件下,AOA与AOB amoA基因拷贝数之比在培养过程中呈先显著升高后降低的趋势(P 0.05)且显著高于4℃、15℃和28℃三个温度条件(P0.05);而4℃条件下AOB和AOA amoA活性均受到抑制。相比于AOB,AOA可能更适应高温环境。     

中性紫色水稻土硝化作用中细菌和古菌的相对贡献

不同的硝化抑制剂对硝化作用的强度以及氨氧化古菌(ammonia-oxidizing archeae,AOA)和氨氧化细菌(ammonia-oxidizing bacteria,AOB)丰度可以产生不同的影响,因此可推算AOA和AOB在硝化作用中的相对贡献。本研究选取硝化过程中AOA和AOB共同起作用的中性紫色土为研究对象,分析不同硝化抑制剂对土壤硝化势以及AOB和AOA的丰度的影响。结果发现,烯丙基硫脲(Allylthiourea,ATU)、2-苯基-4,4,5,5,-四甲基咪唑烷-1-氧-3-氧化物(2-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl3-oxide,PTIO)、辛炔(Octyne、C_8)均可显著降低影响土壤的硝化作用。PTIO显著降低AOA的丰度(约40%),对AOB没有显著影响;ATU和C_8显著降低AOB的丰度(约50%),对AOA没有显著影响。通过计算,该土中AOA与AOB对硝化作用的相对贡献率分别为7.5%和55.6%。结果表明,在中性紫色土中AOA与AOB共同推动硝化作用,其中AOB起主导作用。     

硝化抑制剂对毛竹林土壤N_2O排放和氨氧化微生物的影响

为了探索硝化抑制剂在毛竹生产中的施用技术,通过培养试验研究3,4-二甲基吡唑磷酸盐(DMPP)和双氰胺(DCD)两种硝化抑制剂对毛竹林施用尿素后土壤N2O排放、氮素转化和相关氨氧化细菌(AOB)、氨氧化古菌(AOA)群落结构和丰度的影响。试验设(1)对照(CK)、(2)单施尿素(Urea)、(3)尿素+1%DMPP(DMPP占总N的1%,下同);(4)尿素+1.5%DMPP;(5)尿素+10%DCD;(6)尿素+15%DCD等6个处理,测定N2O的排放动态以及气体排放转折点时的土壤特征指标。结果表明:与单施尿素相比,160 d的时间内两种DMPP用量处理的土壤N2O累积排放减排幅度均为54%,而10%DCD和15%DCD处理的土壤分别减少28%和41%。DMPP和DCD处理50 d和90 d时土壤的NH4+-N含量均显著高于(p0.05)单施尿素处理,而NO3--N含量和表观硝化率则恰好相反,但两种抑制剂间无差异。DMPP处理的AOB群落结构的变化从10 d开始显现,至50 d和90 d时仍保持明显的抑制状态,而DCD处理则至90 d时抑制作用基本消失。单施尿素AOB功能基因(amo A)的丰度均显著高于硝化抑制剂处理(90 d时尿素+10%DCD处理除外);在整个培养期内,尿素和对照土壤的AOA群落结构相似,硝化抑制剂反而增加了AOA功能基因的丰度,表明硝化抑制剂对AOA丰度无明显抑制作用。即两种硝化抑制剂主要通过抑制AOB起作用;调节土壤p H至中性范围,并在1%DMPP施用条件下,硝化抑制剂的抑制效果最显著。     

氮肥施加条件下增温对硝化菌活性和丰度的影响

温度在多种生物地球化学过程中起到关键的调节作用,是影响土壤硝化作用和微生物分布的重要因素之一。硝化过程的第1个步骤由氨氧化细菌(AOB)和氨氧化古菌(AOA)催化,然而,不同施氮量下,增温对硝化菌活性和丰度的影响尚不清楚。本研究基于2008年10月起设立于太行山山前平原的长期增温试验平台(高于地表2m的红外加热器使土壤温度升高1.5℃),于2018年5月对不施氮(N0)和施氮[N1,240kg(N)·hm-2·a-1]下增温分别对0～10 cm和10～20 cm土壤硝化潜势(PNR)、AOA和AOB丰度的影响进行了研究。硝态氮(NO3--N和铵态氮(NH4+-N)含量用分光光度法测量,应用缓冲液培养法测定土壤PNR,提取土壤DNA后用实时荧光定量PCR技术测定功能基因AOA和AOB的丰度。结果表明:温度升高显著增加N1条件下PNR和NO3--N含量(P0.05),降低了N0条件下PNR和NO3--N含量,但差异不显著。N1条件下,增温土壤AOB丰度显著提高(P0.05); N0条件下,增温土壤AOA丰度显著降低(P0.05)。与N0相比, N1条件下的AOA/AOB比值明显降低,表明增温加氮肥处理对AOB的生长刺激更强烈。在增温加施氮条件下,细菌(AOB)表现显著的正反应,在增温不施氮条件下,古菌(AOA)和AOB表现显著的负反应。本研究结果可为全球增温背景下进一步了解硝化活性和氨氧化微生物对增温和氮有效性的响应提供科学依据。     

土地利用方式对万木林土壤氨氧化微生物丰度的影响

以我国亚热带地区典型花岗岩发育酸性红壤为研究对象,选取福建建瓯万木林自然保护区封禁保护下5种自然植被和1种人工种植植被土壤,采用荧光实时定量PCR(Real-time PCR)技术测定了土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)的群落丰度,采用15N稳定同位素成对标记和数值模型相结合的方法测定了土壤初级硝化速率。结果显示,长期封禁保护下的自然植被土壤pH低,土壤AOB数量偏低。人为种植和管理显著提高了土壤pH,促进了AOB的生长,其丰度比自然条件下提高了2个数量级,土壤初级硝化速率也显著提高,并与AOB数量存在显著的相关性,表明AOB是硝化作用的主要贡献者。5种自然植被条件下AOA的amoA基因拷贝数占泉古菌16S rRNA基因的比例都小于1%(0.01%～0.64%),在农业利用方式下上升到5.32%,表明并非所有泉古菌都具备氨氧化功能基因amoA,氮肥施用可能促进了氨氧化古菌的生长。     

短期培养下抑制剂烯丙基硫脲对土壤硝化作用及微生物的影响

硝化抑制剂烯丙基硫脲（ATU）对土壤硝化作用及温室效应的影响及机理尚不清楚。本研究采集典型旱地土壤,进行21天室内微宇宙培养,探究了氮肥与不同剂量ATU（分别为氮素用量的1%, 5%, 10%, 15%和20%）配施对土壤硝化作用及N2O和CO2排放通量的影响,并通过实时荧光定量PCR和高通量测序16S rRNA基因技术监测硝化微生物群落变化,同时与传统硝化抑制剂双氰胺（DCD）进行了保氮减排效果的对比。结果表明,与未施加氮肥的对照相比（CK）,单施氮肥（N）显著提高了土壤硝化强度并促进了N2O排放。DCD能显著抑制硝态氮和N2O的积累,抑制效率分别为68.6%和93.3%。而低浓度ATU对土壤硝化作用无影响,仅在高浓度具有抑制效应,且抑制效率最高仅为14.7%。所有ATU处理N2O排放量均显著降低,降幅为60.3～68.2%,仍远高于DCD处理。处理间N2O和CO2的综合温室效应强弱顺序为N>ATU+N>DCD+N≈CK,且不同ATU施用量处理之间差异不显著。相关分析发现氨氧化细菌（AOB）,而不是氨氧化古菌（AOA）和全程氨氧化细菌（Comammox）,与土壤硝态氮积累和N2O排放显著正相关,与土壤pH显著负相关。高通量测序结果表明Nitrosovibrio tenuis类型AOB对氮肥诱导的硝化过程起主导作用。除此之外,ATU和DCD还能显著提高Cupriavidus,并降低Patulibacter、Aeromicrobium、Actinomycetospora、Defluviicoccus和Acidipila等微生物属在群落中的相对丰度。该研究为深化土壤碳氮循环理论,合理使用硝化抑制剂以及减缓温室气体排放提供科学依据。     

典型旱作农田土壤氧化亚氮排放的氨氧化微生物相对贡献

氨氧化过程对氧化亚氮（N2O）排放具有重要贡献。在不同土壤类型和农田管理下,氨氧化微生物类群对N2O排放的相对贡献组成规律还缺乏系统的研究。本研究选取典型农田耕层土壤（潮土、黑土、砖红壤）,以及有机肥改良的砖红壤剖面土壤,采用选择性抑制法（乙炔和辛炔）研究氨氧化细菌（AOB）、氨氧化古菌和全程硝化菌（AOA+comammox）以及异养硝化菌对土壤硝化潜势、净硝化速率及N2O排放的相对贡献。结果表明,在耕层土壤中,潮土、黑土和砖红壤的pH分别是8.0、6.7和5.7,硝化潜势分别是N 32.5、6.6和4.8 mg?kg-1?d-1,净硝化速率分别是N 7.1、3.0和0.5 mg?kg-1?d-1,7天N2O累积排放量分别是N 38.0、35.4和8.7 μg?kg-1。AOB主导耕层土壤的硝化潜势,对硝化潜势的贡献分别是82%、58%和100%。对于净硝化速率,在潮土和砖红壤中,AOB和AOA+comammox贡献相当（均在30%~40%）,而黑土中由AOB主导（72%）。AOB主导耕层土壤的N2O排放,对N2O排放的贡献分别是72%、92%和58%。在改良的砖红壤剖面土壤中,在0~20 cm、20~40 cm和40~60 cm,pH分别是7.0、5.5和4.9,硝化潜势分别是N 6.6、2.0和1.1 mg?kg-1?d-1,净硝化速率分别是N 4.1、0.9和0.2 mg?kg-1?d-1,N2O排放分别是N 16.3、6.5和2.8 mg?kg-1?d-1。随土壤由深层至表层,硝化潜势、净硝化速率及N2O排放显著提高。AOA+comammox主导表层硝化潜势及净硝化速率的提高（分别贡献63%和54%）,AOB主导N2O排放的增加（贡献54%）。本研究为制定与土壤氨氧化特性及土壤性质相匹配的N2O减排措施提供了新的科学依据。     

脲酶抑制剂与硝化抑制剂对稻田土壤硝化、反硝化功能菌的影响

[目的]在农业生产中,脲酶抑制剂(urease inhibitor,UI)与硝化抑制剂(nitrification inhibitor,NI)常作为氮肥增效剂来提高肥料利用率。本文研究了在我国南方红壤稻田施用脲酶抑制剂与硝化抑制剂后,土壤中氨氧化细菌(ammonia oxidizing bacteria,AOB)、氨氧化古菌(ammonia-oxidizing archaea,AOA)以及反硝化细菌的丰度以及群落结构的变化特征,旨在揭示抑制剂的作用机理及其对土壤环境的影响。[方法]试验在我国南方红壤稻田进行,共设5个处理:1)不施氮肥(CK);2)尿素(U);3)尿素+脲酶抑制剂(U+UI);4)尿素+硝化抑制剂(U+NI);5)尿素+脲酶抑制剂+硝化抑制剂(U+UI+NI),3次重复。脲酶抑制剂与硝化抑制剂分别为NBPT[N-(n-butyl)thiophosphrictriamide,N-丁基硫代磷酰三胺]和DMPP(3,4-dimethylpyrazole phosphate,3,4-二甲基吡唑磷酸盐)。通过荧光定量PCR(Real-time PCR)研究水稻分蘖期与孕穗期抑制剂对三类微生物标记基因拷贝数的影响,并分析土壤铵态氮、硝态氮与三种菌群丰度的相关性;利用变性梯度凝胶电泳(DenaturingGradient Gel Electrophoresis,DGGE)分析抑制剂对土壤AOB、AOA以及反硝化细菌群落结构的影响,并对优势菌群进行系统发育分析。[结果]1)荧光定量PCR结果表明,施用氮肥对两个时期土壤中AOB的amoA基因与反硝化细菌nirK基因的拷贝数均有显著提高,而对AOA的amoA基因始终没有明显影响;AOB与nirK反硝化细菌的丰度与两个时期的铵态氮含量、分蘖期的硝态氮含量呈极显著正相关,与孕穗期的硝态氮含量相关性不显著;DMPP仅在分蘖期显著减少了AOB的amoA基因拷贝数,表明DMPP主要通过限制AOB的生长来抑制稻田土壤硝化过程;NBPT对三类微生物的丰度无明显影响;2)DGGE图谱表明,在分蘖期与孕穗期,施用氮肥均明显增加了图谱中AOB的条带数,而对AOA却没有明显影响;氮肥明显增加了孕穗期反硝化细菌的条带数;与氮肥的影响相比,抑制剂NBPT与DMPP对AOA、AOB以及反硝化菌的群落结构影响甚微;系统发育分析结果表明,与土壤中AOB的优势菌群序列较为接近的有亚硝化单胞菌和亚硝化螺菌。[结论]在南方红壤稻田中,施入氮肥可显著提高AOB与反硝化细菌的丰度,明显影响两种菌群的群落结构,而AOA较为稳定;NBPT对三类微生物的群落结构丰度无明显影响;硝化抑制剂DMPP可抑制AOB的生长但仅表现在分蘖期,这可能是其缓解硝化反应的主要途径;这也说明二者对土壤生态环境均安全可靠。     

亚硝化球菌属(Nitrososphaera)可能是酸性土壤硝化作用的重要驱动者

选择初始pH相近的两个酸性土壤(JX-3和JX-7)样品进行培养试验,探讨了氨氧化古菌(ammonia-oxidizing archaea,AOA)和氨氧化细菌(ammonia-oxidizing bacteria,AOB)在酸性土壤硝化过程中所发挥的作用。结果显示,经过50 d的培养,JX-7样品硝化速率显著高于JX-3,且明显降低土壤pH。培养后,两个土壤样品AOB丰度均增加,但样品间没有显著差异;JX-7土壤AOA丰度显著增加,而JX-3无显著变化。两个土壤样品AOA群落组成本身存在分异,但对于同一样品培养前后均无显著分异;AOB群落组成在两土壤间没有分异,但培养前后分别有分异。培养后,JX-7样品中AOA优势属Nitrososphaera和某些未知微生物的个别OTUs绝对丰度显著增加,而两样品AOB中Nitrosospira属的一些OTUs的绝对丰度均显著增加。因此,所研究的酸性土壤样品中AOA是硝化作用的主要贡献者,而且AOA主要通过提高Nitrososphaera属中个别OTUs的丰度,而不是整个群落来调控硝化作用。     

石灰和双氰胺对红壤酸化和硝化作用的影响及其机制

施用石灰是改良酸性土壤的重要措施,但其对土壤硝化作用的增强不仅加速土壤酸化,也增加硝态氮流失风险.传统的硝化抑制剂双氰胺(Dicyandiamide,DCD)能否在石灰改变pH的条件下始终有效抑制硝化是当前红壤区生产中亟需解决的问题.采用短期土壤培养试验,探讨了不同用量石灰与DCD配合施用对土壤酸化和硝化作用的影响及其...     

Responses of bacterial and archaeal ammonia oxidisers of an acidic luvisols soil to different nitrogen fertilization rates after 9?years

It is still not clear which group of ammonia-oxidizing microorganisms plays the most important roles in nitrification in soils. Change in abundances and community compositions of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) under long-term different nitrogen (N) fertilization rates were investigated in an acidic luvisols soil using real-time polymerase chain reaction and denaturing gradient gel electrophoresis, respectively, based on the ammonia monooxygenase a-subunit gene. The experimental plan included the following treatments: control without N fertilization (N_CK), low N fertilization rate, middle N fertilization rate, and high N fertilization rate as 0, 100, 150, and 250?kg urea-N?ha^?1, respectively. Long-term different N fertilization rates did not significantly alter the total C and N contents of soil while it significantly decreased soil pH, which ranged from 5.60 to 5.20. The AOB abundance was more abundant in the N fertilization treatments than the N_CK treatment; the AOA abundance decreased by the increasing N fertilization rates, as did the ratios of AOA/AOB. The large differences in the potential nitrification rates among four treatments depended on the changes in AOA abundance but not to changes in AOB abundance. Phylogenetic analysis showed that the AOB communities were dominated by Nitrosospira clusters 1, 3, and 9 while all AOA sequences were grouped into soil/sediment cluster except for one sequence. Taken together, these results indicated that AOB and AOA preferred different soil N conditions and AOA were functionally more important in the nitrification than AOB in the acidic luvisols soil.     

Nitrapyrin affects the abundance of ammonia oxidizers rather than community structure in a yellow clay paddy soil

Purpose

Yellow clay paddy soil (Oxisols) is a low-yield soil with low nitrogen use efficiency (NUE) in southern China. The nitrification inhibitor nitrapyrin (2-chloro-6- (tricholoromethyl)-pyridine, CP) has been applied to improve NUE and reduce environmental pollution in paddy soil. However, the effects of nitrapyrin combined with nitrogen fertilizers on ammonia oxidizers in yellow clay paddy soil have not been examined.

Materials and methods

A randomized complete block design was set with three treatments: (1) without nitrogen fertilizer (CK), (2) common prilled urea (PU), and (3) prilled urea with nitrapyrin (NPU). Soil samples were collected from three treatments where CK, PU, and NPU had been repeatedly applied over 5 years. Soil samples were analyzed by quantitative PCR and 454 high-throughput pyrosequencing of the amoA gene to investigate the influence of nitrapyrin combined with nitrogen on the abundance and community structure of ammonia oxidizers in yellow clay paddy soil.

Results and discussion

The potential nitrification rate (PNR) of the soil was significantly correlated with the abundances of both ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB). Application of urea significantly stimulated AOA and AOB growth, whereas nitrapyrin exhibited inhibitory effects on AOA. Phylogenetic analysis showed that the most dominant operational taxonomic units (OTUs) of AOA and AOB were affiliated with the Nitrosotalea cluster and Nitrosospira cluster 12, respectively. AOA and AOB community structures were not altered by urea and nitrapyrin application.

Conclusions

Nitrogen fertilization stimulated nitrification and increased the population sizes of AOA and AOB. Nitrapyrin affected the abundance, but not community structure of ammonia oxidizers in yellow clay soil. Our results suggested that nitrapyrin improving NUE and inhibiting PNR was attributable to the inhibition of AOA growth.

     

Abundance and community structure of ammonia-oxidizing archaea and bacteria in an acid paddy soil

Nitrification is essential to the nitrogen cycle in paddy soils. However, it is still not clear which group of ammonia-oxidizing microorganisms plays more important roles in nitrification in the paddy soils. The changes in the abundance and composition of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) were investigated by real-time PCR, terminal restriction fragment length polymorphism, and clone library approaches in an acid red paddy soil subjected to long-term fertilization treatments, including treatment without fertilizers (CT); chemical fertilizer nitrogen (N); N and potassium (NK); N and phosphorus (NP); N, P, and K (NPK); and NPK plus recycled crop residues (NPK+C). The AOA population size in NPK+C was higher than those in CT, while minor changes in AOB population sizes were detected among the treatments. There were also some changes in AOA community composition responding to different fertilization treatments. Still few differences were detected in AOB community composition among the treatments. Phylogenetic analysis showed that the AOA sequences fell into two main clusters: cluster A and cluster soil/sediment. The AOB composition in this paddy soil was dominated by Nitrosospira cluster 12. These results suggested that the AOA were more sensitive than AOB to different fertilization treatments in the acid red paddy soil.     

长期施用含氯化肥对棕壤硝化作用及氨氧化微生物的影响

【目的】氨氧化微生物是氨氧化过程的主要驱动者,氨氧化过程作为硝化作用的限速步骤对氮循环具有重要作用。本研究以沈阳农业大学棕壤含氯化肥长期定位试验的土壤为研究对象,探讨了连续34年施用高氯和低氯化肥对棕壤硝化作用及氨氧化微生物的影响。【方法】该长期试验在等量氮、磷、钾条件下,设置高氯和低氯处理,共8个处理:T1(不施肥);T2(单施尿素);T3(尿素+氯化钾);T4(尿素+过磷酸钙);T5(尿素+过磷酸钙+氯化钾);T6(尿素+磷酸一铵+氯化钾);T7(尿素+氯磷铵+氯化钾);T8(硝酸磷肥+过磷酸钙+氯化钾),T7为高氯处理。采集0—20cm土壤样品,利用荧光定量PCR技术测定氨氧化细菌(AOB)和古菌(AOA)丰度,并结合土壤硝化潜势和基本化学性质,分析长期施用含氯化肥对棕壤硝化作用及氨氧化微生物丰度的影响及影响氨氧化微生物丰度的主要环境因素。【结果】长期施肥降低了土壤pH值,高氯处理降低得最多,显著低于其他处理;高氯处理的土壤硝化潜势也显著低于其他处理,且除高氯处理外,配施磷肥的处理土壤硝化潜势显著高于不施磷处理。各处理土壤中AOA丰度均显著高于AOB,高氯处理土壤中AOA、AOB丰度均显著低于其他处理,土壤硝化潜势与AOA和AOB均呈显著正相关关系。【结论】连续施用高氯化肥34年显著降低了棕壤AOA和AOB丰度,抑制了硝化潜势。该结果可为通过含氯化肥的合理施用来调节土壤AOA和AOB,进而调控土壤氮素循环提供参考。     

Responses of soil nitrous oxide production and abundances and composition of associated microbial communities to nitrogen and water amendment

Soil moisture and nitrogen (N) are two important factors influencing N₂O emissions and the growth of microorganisms. Here, we carried out a microcosm experiment to evaluate effects of soil moisture level and N fertilizer type on N₂O emissions and abundances and composition of associated microbial communities in the two typical arable soils. The abundances and community composition of functional microbes involved in nitrification and denitrification were determined via quantitative PCR (qPCR) and terminal restriction length fragment polymorphism (T-RFLP), respectively. Results showed that N₂O production was higher at 90% water-filled pore (WFPS) than at 50% WFPS. The N₂O emissions in the two soils amended with ammonium were higher than those amended with nitrate, especially at relatively high moisture level. In both soils, increased soil moisture stimulated the growth of ammonia-oxidizing bacteria (AOB) and nitrite reducer (nirK). Ammonium fertilizer treatment increased the population size of AOB and nirK genes in the alluvial soil, while reduced the abundances of ammonia-oxidizing archaea (AOA) and denitrifiers (nirK and nosZ) in the red soil. Nitrate addition had a negative effect on AOA abundance in the red soil. Total N₂O emissions were positively correlated to AOB abundance, but not to other functional genes in the two soils. Changed soil moisture significantly affected AOA rather than AOB community composition in both soils. The way and extent of N fertilizers impacted on nitrifier and denitrifier community composition varied with N form and soil type. These results indicate that N₂O emissions and the succession of nitrifying and denitrifying communities are selectively affected by soil moisture and N fertilizer form in the two contrasting types of soil.     

冬种绿肥对水稻土硝化作用的影响

冬闲田种植绿肥是传统的水稻土培肥增产措施,但绿肥-水稻种植系统中,不同绿肥种类对硝化作用的影响规律及调控机制尚不明确.采用盆栽试验,研究了冬种紫云英、油菜、黑麦草对土壤性状及硝化作用的影响,并通过特异性细菌抑制剂(卡那霉素和大观霉素)研究了氨氧化细菌(AOB)和氨氧化古菌(AOA)对硝化作用的相对贡献.结果表明,冬种三...