长期定位施肥对中性紫色土硝化作用及氨氧化微生物的影响

为了研究长期不同施肥措施对中性紫色土氨氧化微生物及其硝化作用的影响,以国家紫色土肥力与肥料效益监测基地的中性紫色土为研究对象,进行土壤氨氧化细菌和氨氧化古菌amo A基因的Real-time PCR分析,比较长期不同定位施肥对土壤氨氧化潜势和硝化强度的影响,并分析不同施肥制度对功能微生物丰度与功能的作用。数据显示,土壤中氨氧化古菌amo A基因拷贝数(Log值6.21～7.14)远大于氨氧化细菌(Log值3.65～5.73),相对于对氨氧化细菌丰度的影响,施肥对土壤氨氧化古菌丰度影响较小。施用氮肥与磷肥都显著提高了土壤氨氧化细菌丰度,1.5NPK+M处理氨氧化细菌丰度最高(Log值5.73),有机无机肥配施可以显著提高土壤氨氧化微生物丰度;而含氯化肥的施用在一定程度上降低了土壤氨氧化细菌丰度与硝化细菌生长,与施用不含氯的肥料处理相比,含氯肥料处理的土壤氨氧化细菌丰度与硝化细菌数分别降低了3.74%和88.12%。研究表明,长期施肥能影响中性紫色土中氨氧化细菌的丰度,有机无机肥配施能够提高土壤的氨氧化潜力与土壤的硝化能力。     

氮肥水平对稻田细菌群落及N2O排放的影响

作为土壤氮素转化的驱动者,微生物群落结构关系着稻田氮素利用及温室气体N_2O排放等问题。本研究分别基于高通量测序和荧光定量PCR技术,分析了不同氮肥水平[CK(不施氮)、N(施N 180 kg·hm-2)、2/3N(施N 120 kg·hm-2)、1/3N(施N 60 kg·hm-2)]下稻田细菌群落及硝化反硝化关键微生物功能基因丰度的变化。结果显示:氮肥水平提高增加了稻田细菌物种丰富度Chao1指数和群落多样性Shannon指数,改变了细菌群落组成,其中与硝化作用相关的硝化螺菌门Nitrospirae和嗜酸的醋杆菌门Acidobacteria的相对丰度随氮肥水平提高而增加,但甲烷氧化菌Methylosinus的相对丰度随氮肥水平提高而降低。氮肥水平对稻田硝化作用关键微生物氨氧化细菌amo A基因丰度的影响较大,0~5 cm和10~20 cm深度土层中的amo A基因丰度均随氮肥用量增加而提高;反硝化作用关键微生物功能基因nir S、qno B和nos Z的丰度在不施肥处理(CK)中显著低于施肥处理(1/3N、2/3N和N)(P0.05),但1/3N、2/3N和N处理的稻田nir S基因丰度没有明显差异;0~5 cm土层中qno B和nos Z基因丰度存在随氮肥水平提高而增加的趋势,10~20 cm土层中nos Z基因丰度在2/3N和N处理下显著高于1/3N处理(P0.05)。N处理的稻田N_2O排放通量显著高于2/3N及1/3N处理(P0.05),后者又显著高于CK处理(P0.05)。相关分析结果表明稻田N_2O排放通量与0~5 cm土层中硝化螺菌门Nitrospirae相对丰度及10~20 cm土层中amo A基因丰度存在显著相关性(P0.05,n=10)。综上所述,氮肥水平提高增加了稻田细菌群落多样性,促进了稻田N_2O排放,且本研究稻田中硝化作用微生物群落及丰度变化与稻田N_2O排放的关系更为密切。     

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。     

转Bt基因棉秸秆还田利用对土壤肥力的影响

随着转基因的快速发展,大量转Bt棉秸秆的合理利用和处理是不可忽视的重要课题之一。为明确Bt棉秸秆还田利用的可行性和安全性,本研究以不同抗虫转Bt基因棉和常规棉花‘泗棉3号’为研究材料,在分别种植1、2年后将秸秆机械粉碎后原位还田,测试土壤中Bt蛋白残留量、土壤酶活性及养分含量的变化,分析Bt棉秸秆原位还田对土壤肥力特性的影响。研究结果表明,秸秆还田40 d后, Bt棉样地土壤中Bt残留蛋白检测值较低,均与非转基因棉样地无显著性差异。棉秸秆还田后,土壤脲酶、蔗糖酶、蛋白酶、多酚氧化酶、过氧化氢酶、碱性磷酸酶活性皆较秸秆还田前增加,但土壤纤维素酶活性较之前降低。棉秸秆还田使土壤中有机质、有效磷、碱解氮、速效钾和全氮等养分含量及pH明显增加,而Bt抗虫棉与常规棉秸秆还田后对土壤肥力的影响不存在显著差异。对土壤综合肥力指数评价结果表明,秸秆还田对土壤肥力提升与Bt棉抗虫水平无关,土壤肥力指数在两年间由Ⅲ级水平上升至Ⅱ级水平。综上, Bt棉花秸秆还田不会造成土壤综合肥力降低,相反能有效提升土壤肥力;同时还田利用措施可对转基因植株有效灭活,与转基因植物秸秆利用和无害化处理要求相契合。生产中用于Bt转基因棉花秸秆利用和处理在一定程度上是安全可行的。     

稻虾共作对稻田土壤nirK反硝化微生物群落结构和多样性的影响

稻虾共作是水稻种植与克氏螯虾共作形成的互利共生的稻田种养复合生态模式。目前对稻虾共作模式稻田反硝化微生物多样性和群落结构的影响尚不清楚。本研究以江汉平原常规中稻模式(MR)为对照,设置连续3年(2014—2016年)稻虾共作模式(CR)为处理,通过特异引物提取中稻抽穗期稻田土壤nirK基因,采用Illumina Miseq高通量测序技术,探讨稻虾共作模式对稻田土壤nirK反硝化微生物多样性和群落结构的影响。结果表明:稻虾共作模式显著提升水稻抽穗期稻田土壤中硝态氮、全氮及全碳的含量,对土壤碳氮比、碱解氮和铵态氮含量没有显著影响。稻虾共作模式显著增加稻田土壤nirK基因微生物的丰富度指数,但对nirK基因微生物的多样性指数影响不显著。稻虾共作模式改变了nirK基因微生物在目、科、属、种水平的群落组成,较常规中稻模式,稻虾共作模式在各分类水平组成类群均减少;稻虾共作模式较常规中稻模式改变了目的种类,对共有目相对丰度没有显著性改变。RDA分析表明稻虾共作模式对土壤nirK基因菌群的群落结构有一定的改变,但稻虾共作模式与常规中稻模式在群落结构上仍保留着一定的相似性。硝态氮含量是影响nirK反硝化细菌群落结构的主效因子。可见,稻虾共作模式对微生物多样性指数没有显著影响,但显著增加了微生物丰富度指数,改变了稻田土壤nirK反硝化微生物在目、科、属、种的群落结构。     

不同施肥模式对土壤氮循环功能微生物的影响

  【目的】   微生物在土壤氮循环过程中发挥着重要作用。通过研究农田土壤氮循环过程中不同功能微生物群落基因丰度对施肥模式的响应及其关键影响因素,探讨不同施肥模式调控下氮素转化的微生物学机制,为改善农业生产中氮素的管理策略提供理论依据。   【方法】   田间试验始于2011年,试验地点位于江苏省常州市溧阳市南渡镇,供试土壤为白土型水稻土,种植制度为稻麦轮作。试验包括单施化肥 (NPK)、化肥+畜禽有机肥 (NPKM)、化肥+秸秆还田 (NPKS) 以及相邻江苏省耕地质量监测点不施肥对照 (CK),共 4个处理。于2014年水稻成熟期采集土壤样品,采用实时荧光定量PCR法分析了土壤硝化 (amoA)、反硝化 (narG、nirS、nirK、norB、nosZ)、固氮 (nifH)、硝酸盐异化还原 (napA) 等氮循环过程的相关功能微生物基因丰度的变化。以氨氧化微生物为模式微生物,测定添加与不添加1-辛炔情况下的土壤硝化潜势,分析氨氧化古菌 (AOA) 与氨氧化细菌 (AOB) 功能基因丰度与土壤硝化功能的内在联系。   【结果】   与CK相比,NPK处理显著增加了土壤中AOB-amoA、narG、nosZ和nifH基因的丰度。与NPK处理相比,NPKS处理进一步提高了土壤中AOB-amoA、narG、nosZ、nifH以及nirK基因的丰度。与CK相比,除AOA-amoA、nirS、napA基因以外,NPKM处理显著提高了土壤中所有氮循环功能基因的丰度。AOB-amoA基因丰度的变化对土壤氮循环功能基因丰度的整体变异影响最大。AOB主导了施肥土壤的硝化过程 (81.90%～84.42%)。土壤总硝化潜势与AOB-amoA基因丰度显著相关,但与AOA-amoA基因丰度相关性不显著。氮循环功能微生物基因丰度主要受到土壤pH、土壤有机碳(SOC)和NO₃–含量的影响。   【结论】   畜禽有机肥与秸秆的施用能够进一步刺激氮循环功能基因丰度的增长,促进土壤氮循环。土壤pH、SOC和NO₃–含量是影响土壤氮循环功能微生物丰度的关键因素。施肥主要通过提高土壤AOB-amoA功能基因的丰度,进而提高土壤硝化潜势,因此在控制土壤硝化作用时应重点关注AOB微生物群落。     

稻田土壤厌氧氨氧化菌群落结构对长期不同施肥的响应

研究长期不同施肥稻田土壤厌氧氨氧化微生物丰度和群落结构组成,深入认识稻田厌氧氨氧化菌对不同施肥的响应机理,可为合理施肥和理解湿地生态系统厌氧氨氧化过程提供科学依据。设置不施肥(CK)、单施无机肥(NPK)、无机肥配施牛粪(NPKM)、无机肥加秸秆还田(NPKS)四个处理,采用荧光定量PCR和高通量测序对不同施肥模式下水稻土厌氧氨氧化细菌丰度和群落结构进行分析。结果发现,不同处理之间厌氧氨氧化细菌丰度具有显著差异(p0.05),表现为NPKMNPKSNPKCK,且与有机质、全氮和铵态氮含量具有显著相关性(p0.05)。高通量测序结果表明,不同施肥处理主要的厌氧氨氧化菌为Candidatus Brocadia、Candidatus Anammoxoglobus和Candidatus Scalindua,其中优势种群为Candidatus Brocadia。菌群的多样性分析表明,CK和NPKS处理的厌氧氨氧化菌群落结构多样性香农指数(Shannon index)、辛普森指数(Simpson index)和丰富度指数(Chao 1 index)显著高于NPKM和NPK处理(p0.05)。上述结果表明,长期施肥改变了厌氧氨氧化菌的数量和群落结构。有机无机肥配施更有利于提高厌氧氨氧化菌丰度。然而,不同施肥措施对厌氧氨氧化菌的多样性影响不同,无机肥加秸秆还田提高了厌氧氨氧化菌多样性,但单施无机肥和无机肥配牛粪降低了厌氧氨氧化菌多样性。厌氧氨氧化菌的数量和群落结构对不同施肥的响应不同。     

转Cry1Ab/Ac基因水稻对稻田底栖动物群落的影响

转Bt基因抗虫水稻的成功研发,为有效控制鳞翅目害虫对水稻的危害提供了一条最为有效、经济的途径。稻田底栖动物群落是转Bt基因抗虫水稻的重要非靶标生物。为了解转Bt基因水稻对稻田底栖动物群落的安全性,以转Cry1Ab/Ac基因抗虫水稻‘华恢1号’(简写为HH1)为材料,以其非转基因亲本‘明恢63’(简写为MH63)为对照,以稻田底栖动物群落为指示生物,运用群落学方法,通过连续2年在江西南昌的大田试验,研究了转Cry1Ab/Ac基因抗虫水稻对稻田底栖动物群落的影响。结果显示:2012年在HH1与MH63的稻田中分别采集到底栖动物22种与25种,其中两生境中相同物种19种,两群落物种组成相似度为0.808 5,有7种优势种,优势种相似度为0.833 3。2013年在HH1与MH63稻田中分别采集到底栖动物26种与28种,两类稻田相同底栖动物22种,两群落物种组成相似度为0.814 8,有6种优势种,优势种相似度为1.000 0。2012年与2013年群落总体分析与时间动态分析均显示,两类稻田中底栖动物的物种丰富度、个体数量、多样性指数、均匀性指数及优势集中性指数变化趋势相似,且均无显著差异。结果表明:连续种植2年转Cry1Ab/Ac基因水稻HH1对南昌稻田底栖动物群落无明显的负作用。     

长期施肥对稻田土壤微生物群落结构及氮循环功能微生物数量的影响

  【目的】   稻田是陆生生态系统中重要的氮库之一，在氮素生物地球化学循环中具有重要地位。研究不同施肥处理对稻田土壤微生物群落结构及其功能的影响具有重要意义。   【方法】   田间试验位于江苏省金坛市，在取样时试验已进行了6年。施肥处理包括:不施肥对照 (CK)、施化肥 (CF)、化肥+猪粪混施 (CMF)、化肥+秸秆混施 (CSF)。采用高通量测序和定量PCR方法测定稻田土壤微生物群落结构及氮循环相关功能微生物数量。   【结果】   在施用肥料6年后，土壤全碳、可溶性有机碳、全氮、铵态氮和硝态氮含量均不同程度地提高。与CF相比，CSF和CMF处理土壤pH升高，全碳、可溶性有机碳与养分含量升高。CK与施肥处理的土壤细菌群落结构差异明显，不同施肥处理的细菌群落结构之间有明显差别。聚类结果显示，CK与CMF处理细菌群落聚类更接近，CF处理和CSF处理细菌群落结构更为接近；与CK相比，CF、CMF、CSF处理土壤中氨氧化细菌 (AOB) 和铁氨氧化微生物Feammox A6的丰度显著提高，其中Feammox A6分别增长87.6%、158%和157%。冗余分析结果表明，施肥过程及其对土壤化学性质的改变显著影响土壤细菌群落的组成和分布。   【结论】   施肥导致的反应底物 (NH₄+、NO₃–含量) 及土壤理化性质的差异，是土壤微生物群落结构和功能微生物数量响应的主要决定因素。不施肥与化肥配施猪粪的土壤细菌群落聚类更接近，施化肥与化肥配施秸秆的细菌群落结构更为接近。施肥对氨氧化细菌AOA数量影响不明显，但显著提高氨氧化古菌AOB和厌氧铁氨氧化功能微生物Feammox A6的数量，特别是有机肥 (猪粪、秸秆) 提高Feammox A6数量的效果大于化肥。长期单施化肥土壤中厌氧氨氧化细菌丰度显著降低，反硝化功能基因nirK、nosZ丰度显著增高；化肥配施猪粪土壤中的厌氧氨氧化细菌丰度变化不明显，反硝化功能基因narG、nirK、nosZ丰度显著增高；化肥配施秸秆处理厌氧氨氧化细菌丰度变化不明显，反硝化功能基因nirK、nosZ丰度显著增高。     

水稻土厌氧氨氧化对不同耕作方式与秸秆还田的响应

【目的】研究稻田土壤厌氧氨氧化微生物活性、丰度和群落结构对不同耕作方式与秸秆还田的响应,为稻田土壤氮素管理提供科学依据。【方法】设置3个不同耕作方式(翻耕、旋耕、免耕)+秸秆还田处理,同时设置翻耕+秸秆不还田处理作为对照,采用_¹⁵N示踪法、荧光定量PCR及Illumina测序分别分析厌氧氨氧化菌潜在活性、功能基因hzsB拷贝数及群落组成。【结果】秸秆还田条件下,厌氧氨氧化菌潜在活性表现为免耕>旋耕>翻耕(p<0.05),对照与旋耕+秸秆还田处理无显著性差异。各处理之间厌氧氨氧化菌功能基因(hzsB)拷贝数无显著性差异。Illumina测序从属水平鉴定出Kuenenia和Scalindua两种已知的厌氧氨氧化菌及部分未鉴定属,且各处理间二者的相对丰度具有负相关关系(p<0.05)。相关性分析结果表明,厌氧氨氧化菌活性与土壤反硝化活性显著相关,与功能基因拷贝数及群落组成无显著相关性。【结论】水稻田长期秸秆还田和不同耕作方式改变了厌氧氨氧化菌活性和群落组成,但其数量未发生显著变化。水稻田秸秆还田加翻耕可一定程度上抑制土壤氮损失,而秸秆还田与免耕结合的田间管理方式提高了土壤氮素损失潜力。     

A new mycorrhizal helper bacterium, <Emphasis Type="Italic">Ralstonia</Emphasis> species,in the ectomycorrhizal symbiosis between <Emphasis Type="Italic">Pinus thunbergii</Emphasis> and <Emphasis Type="Italic">Suillus granulatus</Emphasis>

In a Robinia-pseudoacacia-dominated coastal forest in Tottori prefecture Japan, the growth and survival of Pinus thunbergii seedlings and the natural regeneration of P. thunbergii was disturbed by R. pseudoacacia. In order to improve the growth of P. thunbergii seedling in the Tottori sand dune, we tried to find a mycorrhiza helper bacteria (MHB) from P. thunbergii mycorrhizosphere in a Tottori sand dune. Two MHB, Ralstonia sp. and Bacillus subtilis, were selected from the nine bacterial species isolated from the mycorrhizosphere of P. thunbergii. The bacterial effect on the ectomycorrhizal fungus Suillus granulatus was investigated by confrontation assay and a microcosm experiment. The confrontation assay showed that Ralstonia sp. promoted the hyphal growth of S. granulatus. Moreover, the S. granulatus–P. thunbergii symbiosis was significantly stimulated by Ralstonia sp. and B. subtilis. Ralstonia sp. and B. subtilis were regarded as MHB associated with P. thunbergii. This is the first report of Ralstonia sp. as an MHB.     

Effects of herbicide butachlor application on the growth of periphytic biofilms and nitrogen loss in paddy systems

The application of butachlor as an herbicide in paddy fields is widely practiced,aiming to increase rice yield by directly or indirectly influencing the paddy environment.Periphytic biofilms,which form at the soil-water interface in paddy fields,are complex bioaggregates that play an important role in nitrogen (N) cycling.The objective of this study was to investigate the effect of butachlor on periphytic biofilm growth and N cycling under both light and dark conditions in the laboratory.The res...     

两株解磷细菌的解磷活性及作用机制研究

解磷细菌在增加土壤可溶性磷含量、提高磷肥利用效率方面具有重要作用。为选筛高效解磷菌、探讨其解磷机制,本文利用平板溶磷圈法筛选解磷细菌,采用钼锑抗比色法研究其解磷活性,苯磷酸二钠法研究其磷酸酶活性,利用薄层层析分析其产生的有机酸,根据生理生化特征和16S r RNA基因序列系统发育分析,确定其分类学地位。结果表明,菌株JXJ-11和JXJ-15对植酸钙的降解活性很强,3 d后培养液中可溶性磷浓度分别增加219 mg·L~(-1)和216 mg·L~(-1);对磷酸钙降解活性较弱,最高可溶性磷浓度仅为植酸钙的21.79%~30.37%;解磷细菌可分泌酸性、中性和碱性磷酸酶,降解不溶性磷,可能产生丙酸和琥珀酸等有机酸,降低培养液p H,增加可溶性磷浓度。两株细菌均为革兰氏阴性杆菌,无芽孢,产生硫化氢,其中菌株JXJ-11的16S rRNA基因序列与Sphingomonas melonis DAPP-PG 224T和S.aquatilis JSS7T相似性最高(99.79%),菌株JXJ-15的16S rRNA基因序列与Klebsiella pneumoniae subsp.pneumoniae DSM 30104T相似性最高(99.73%),根据以上信息,确定菌株JXJ-11和JXJ-15分别是鞘氨醇单胞菌属和克雷白氏杆菌属的成员。菌株JXJ-11和JXJ-15的解磷机制包括分泌有机酸和磷酸酶,其中JXJ-11在微生物磷肥研制方面具有潜在应用价值。     

Mitigation of salt stress in wheat seedlings by a <Emphasis Type="Italic">gfp</Emphasis>-tagged <Emphasis Type="Italic">Azospirillum lipoferum</Emphasis>

Root colonization and mitigation of NaCl stress on wheat seedlings were studied by inoculating seeds with Azospirillum lipoferum JA4ngfp15 tagged with the green fluorescent protein gene (gfp). Colonization of wheat roots under 80 and 160 mM NaCl stress was similar to root colonization with this bacterial species under non-saline conditions, that is, single cells and small aggregates were mainly located in the root hair zone. These salt concentrations had significant inhibitory effects on development of seedlings, but not on growth in culture of gfp-A. lipoferum JA4ngfp15. Reduced plant growth (height and dry weight of leaves and roots) under continuous irrigation with 160 mM NaCl was ameliorated by bacterial inoculation with gfp-A. lipoferum JA4ngfp15. Inoculation of plants subjected to continuous irrigation with 80 mM NaCl or to a single application of either NaCl concentration (80 or 160 mM NaCl) did not mitigate salt stress. This study indicates that, under high NaCl concentration, inoculation with modified A. lipoferum reduced the deleterious effects of NaCl; colonization patterns on roots were unaffected and the genetic marker did not induce undesirable effects on the interaction between the bacterium and the plants.     

微生物介导的农业土壤氧化亚氮减排研究进展

氧化亚氮(N₂O)是一种在大气存留时间长且破坏臭氧层的重要温室气体。农业土壤源N₂O是其重要来源,具有产生路径广、影响因素多、调控复杂等特点。减少农业土壤N₂O排放一直是研究的热点。含有N₂O还原酶的N₂O还原细菌能将N₂O还原为氮气(N2),这是目前已知的微生物还原N₂O唯一的汇。直接应用微生物减少农业土壤N₂O排放是一种新兴的减排技术。本文详细阐述了农业土壤N₂O的生物源与汇,重点论述了N₂O减排微生物的筛选及应用策略。综述了微生物介导的农业土壤N₂O减排的两种微生物生态学机制：一种是利用含有nos Z基因的N₂O还原细菌直接减少N₂O排放,另一种是利用能改变N₂O还原细菌群落组成和丰度及其活性的植物根际促生菌间接减少N₂O排放。最后,讨论了影响微生物介导的...     

Light-stimulated 14CO2 uptake and acetylene reduction by bacteriochlorophyll containing stem nodule isolate BTAi 1

Summary Bradyrhizobial strain BTAi 1 nodulates both stems and roots of Aeschynomene spp. Previous work has shown that it contains bacteriochlorophyll a and forms photosynthetic reaction centers, and has provided indirect evidence of photosynthesis by bacteroids within stem nodules. Here we report physiological and biochemical characteristics of BTAi 1 ex planta, which also suggest the presence of photosynthetic activity. Light-stimulated uptake of ¹⁴CO₂ by BTAi 1 was detected at all stages of growth. Inhibitors of photosynthesis, 1,10-orthophenanthroline and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), and the uncoupler NH₄Cl, immediately suppressed light-driven ¹⁴CO₂ uptake and increased O₂ uptake. BTAi 1 is strictly aerobic and was unable to grow without organic C even in the light; also, it was unable to grow chemoautotrophically in an atmosphere enriched with H₂ and CO₂. In micro-aerobic conditions, strain BTAi 1 expressed acetylene reducing activity ex planta in an N-free medium. The highest rates of light-stimulated ¹⁴CO₂ uptake and acetylene-reducing activity occurred during the exponential and early stationary phases of growth. Acetylene-reducing rates at a low glucose concentration were increased following a light-dark cycle in comparison with continuous dark conditions.     

应用新引物DGGE分析土壤中硝酸盐还原细菌群落

The narG gene is frequently used as a molecular marker for bacterial nitrate-reducing community analysis. In this study, a new set of primers targeting the narG gene was designed and applied to semi-nested polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) assay. The potential of the new primers was verified on DNA directly extracted from soils from five different experimental sites distributed in Central and Southern Italy. Specificity of the primers was determined by excision, amplification, and sequencing of bands resolved by DGGE. A phylogenetic analysis showed the correlation between the sequences retrieved from the soils studied and the narG sequences from β and γ-Proteobacteria. These primers expanded the existing molecular tools for ecological study on the size and diversity of nitrate-reducing bacterial community in soil.     

三种农田土壤中氨氧化细菌amoA基因多样性比较分析

比较分析中国东部不同季风气候区中海伦黑土(HL)、封丘潮土(FQ)和鹰潭红壤(YT)3种土壤氨氧化细菌amoA基因多样性。采用非培养方法直接从土壤中提取微生物总DNA,用氨氧化细菌amoA基因特异引物扩增总DNA,构建了3种土壤amoA基因文库,并对文库进行限制性长度多态性(RFLP)分析。HL、FQ和YT的amoA基因文库克隆数量分别为49、50和48个,相应的RFLP类型数为10、10和14个OTUs,其中有4个OTUs为三种土壤共有;YT中氨氧化细菌amoA基因多样性指数最高,FQ最低;HL和FQ群落的相似为70%,HL与YT的相似度为50%,而FQ和YT之间仅为42%,说明氨氧化细菌具有地理分布的规律:17个amoA基因序列可以被聚成6个cluster,分属Nitrosospira和Nitrosomonas两个属。三种农田土壤中均存在丰富的氨氧化细菌,表明氨氧化细菌在农田土壤氮素循环中具有重要作用。     

Powdery mildew resistance in Aegilops tauschii coss. and synthetic hexaploid wheats

Summary A collection of 400 Ae. tauschii (syn. Ae. squarrosa) Coss. accessions were screened for powdery mildew resistance based on the response patterns of 13 wheat cultivars/lines possessing major resistance genes to nine differential mildew isolates. 106 accessions showed complete resistance to all isolates, and 174 accessions revealed isolate-specific resistance, among which were 40 accessions exhibiting an identical response pattern as wheat cultivar Ulka/^*8Cc which is known to possess resistance gene Pm2. Expression of both complete and isolate-specific resistance from Ae. tauschii was observed in some synthetic hexaploid wheats derived from four mildew susceptible T. durum Desf. parents, each crossed with five to 38 resistant diploid Ae. tauschii accessions. Synthetic amphiploids involving different combinations of T. durum and Ae. tauschii generally showed a decrease in resistance compared with that expressed by the Ae. tauschii parental lines.     

Agricultural potential of Mediterranean grain and forage legumes: Key differences between and within Vicia species in terms of phenology, yield, and agronomy give insight into plant adaptation to semi-arid environments

In order to identify key adaptive traits which affect productivity in Mediterranean grain and forage legumes and simultaneously determine the agricultural potential of a wide range of Vicia species, germplasm collected from the wild throughout the eastern Mediterranean was grown under semi-arid conditions in Tel Hadya, Syria (313 mm growing season rainfall). These included species currently in use in Mediterranean agriculture, such as V. sativa L., as well as those more widely used in the past-such as V. ervilia L., but also a broad selection from Section Narbonensis (B. Fedtsch. ex Radzhi) Maxted, including V. narbonensis L, V. johannis Tamamsch., V. hyaeniscyamus Mout., V. serratifolia Jacq., and V. kalakhensis Khattab et al. V. faba, a near relative of the taxa in Section Narbonensis, was included as a domesticated control. Where applicable, a representative range of subspecies was used. Accessions were chosen from a wide range of habitats in terms of latitude (31.02–40.72 decimal^o), longitude (27.1–43.17 decimal^o), altitude (20–1510 m), rainfall (180–1700 mm/yr) and soil depth (5–50 cm) in order to maximise diversity within species. Agricultural potential was determined by measuring seed, hay and biological yield, as well as agronomic traits such as harvest index, standing crop height, and seed size. The comparative influence of phenology and key agronomic traits such as plant habit and seed size on productivity varied tremendously between species, depending on their reproductive strategies. In V. sativa and V. ervilia, the smaller seed species which rely on long vegetative phases and growing seasons to accumulate sufficient biomass to set seed, and in which there was comparatively little agronomic variation, phenology had a large impact on yield. In early emerging taxa such as V. ervilia and V. s. subsp. sativa, with built-in long vegetative phases and growing seasons, seed yield was negatively correlated with flower ing (r = –0.86 to –0.88), whereas the opposite was the case for later emerging taxa such as V. s. subsp. nigra (L.) Ehrh. (r = 0.95). Within V. narbonensis and relatives, the larger seeded Vicia species which rely on more conservative reproductive strategies where high seedling vigour associated with large seeds enables the species to enter reproductive phases relatively early, phenology had a much smaller impact on yield than did variation of key traits such as seed weight, plant habit and pod shattering. Among the undomesticated germplasm harvest indices ranged from 0.09–0.31, hay yields from 0.1–3.4 t/ha, seed yield from 0–2.0 t/ha, and dry matter at maturity from 1.6–6.5 t/ha. Sub-specific taxonomy was crucial in determining agronomic potential. V. narbonensis var. aegyptiaca Kornhuber ex Asch. et Schweinf. showed the most potential, combining an upright habit, large seeds (212 mg) and tendency to retain intact pods after maturity, with the highest yield, harvest index and crop height of all the wild Vicia species. V. sativa subsp. sativa, V. ervilia and V. narbonensis var. narbonensis were less productive, but still showed agricultural potential. The smaller seeded V. narbonensis, var. affinis, var. jordanica H. Schäf. and var. salmonea (Mout.) H. Schäf., and their close relatives V. johannis, V. hyaeniscyamus, V. serratifolia and V. kalakhensis have little to offer Mediterranean agriculture on the basis of poor agronomy.