Daily dynamics of the number and activity of nitrogen-fixing bacteria in fallow and intensely cultivated soils

The daily dynamics (during 33 days) of the number (colony-forming units (CFU)) of nitrogen-fixing bacteria and of the nitrogen-fixing activity (the acetylene method) were determined in a gray forest soil under a fallow land and under an intensely cultivated field. The daily dynamics of the CFUs determined on the nitrogen-free medium in the samples from both plots had wavelike patterns. The daily values of the actual and potential activities of nitrogen fixation in the samples from the fallow land plot and of the actual activity of nitrogen fixation in the samples from the intensely cultivated soil were low and close to the detection limit. The potential activity of nitrogen fixation in the intensely cultivated soil was significant and also had a wavelike pattern. The harmonic analysis of the daily dynamics of the CFUs and nitrogen fixation showed the statistically significant harmonics of these biological characteristics pointing to the objective and regular character of the wavelike dynamics. The revealed dynamics of the biological characteristics of the soils and the methods of their analysis are important in terms of the comparative study of the biological properties of different soils.     

Stimulation of nitrogen fixation in soddy-podzolic soils with fungi

Stimulation of nitrogen fixation in soddy-podzolic soils is related to the hydrolytic activity of fungi decomposing plant polymers. It was found that the rate of nitrogen fixation upon the simultaneous inoculation of the strains of nitrogen-fixing bacteria Bacillus cereus var. mycoides and the cellulolytic fungus Trichoderma asperellum into a sterile soil enriched with cellulose or Jerusalem artichoke residues is two to four times higher than upon the inoculation of the strains of Bacillus cereus var. mycoides L1 only. The increase in the nitrogen fixation depended on the resistance of the substrates added into the soil to fungal hydrolysis. The biomass of the fungi decomposing plant polymers increased by two–four times. The nitrogen-fixing activity of the soil decreased when the growth of the fungi was inhibited with cycloheximide, which attested to a close correlation between the intensity of the nitrogen fixation and the decomposition of the plant polymers by fungi. The introduction of an antifungal antibiotic, together with starch or with plant residues, significantly (by 60–90%) decreased the rate of nitrogen fixation in the soll.     

Microbial transformation of nitrogen compounds in middle taiga soils

The intensity of mineralization, nitrogen fixation, and denitrification in forest soils of the Karelian middle taiga ecosystems has been evaluated. Podzol-gleyish soil underlying a birch forest with gramineous plants and miscellaneous herbs was shown to have the highest nitrogen-fixing activity. The loss of gaseous nitrogen during denitrification was insignificant due to the low nitrifying activity of the soils named above. N₂O uptake by microorganisms was rather intensive in all the soils analyzed, and in illuvial-humo-ferric podzols underlying pine and spruce forests this process predominated. Podzolic sandy loam gley-like soil of a birch forest with gramineous plants and miscellaneous herbs had the highest potential for the mineralization of organic nitrogen; the rate of ammonification and nitrification in this soil was maximal.     

固氮芽孢杆菌N3的筛选鉴定及其对二月兰的促生效果

从山东泰安农田土壤中筛选获得1株固氮能力强的菌株N3,通过形态观察、生理生化特征以及16S rDNA基因序列分析,确定为巨大芽孢杆菌属(Bacillus megaterium),该菌株固氮酶活性达C2H428.33 nmol/(h·ml)。温室条件下进行二月兰盆栽试验,设置不接菌对照(CK)、接种巨大芽孢杆菌N3、接种产吲哚乙酸(IAA)菌CDL29和复合接种菌株N3+CDL29四个处理,探讨了接种不同植物促生菌对二月兰的促生效果。结果表明:接种植物促生菌均促进二月兰的生长,且单一接种固氮菌株N3的促生效果最好,其显著提高二月兰地上部氮、磷和地下部氮、钾的含量,相比对照,增幅分别为33.03%、55.56%和29.87%、51.11%,其他处理也有提高二月兰养分含量的趋势,但并不显著;单一接种菌株N3,土壤有机质、全氮和速效养分含量显著提高;接种植物促生菌均提高土壤微生物生物量碳(MBC)和氮(MBN)的含量,单一接种固氮菌株N3提升效果最佳,接种产IAA菌株CDL29和复合接种次之。此外,接种固氮菌株N3能够显著提升土壤固氮酶活性,接种菌株CDL29对土壤固氮酶活性没有显著影响。因此,单一接种固氮菌株N3在促进二月兰生长、养分含量提高及土壤理化和生物学性质改良上都具有显著的效果。     

Atmospheric nitrogen fixation by photosynthetic microorganisms in a submerged Philippine soil

Photosynthetic nitrogen-fixing microorganisms help maintain the nitrogen level of soil in rice paddies when environmental factors favor the growth of microorganisms. Our studies showed that blue-green algae in particular have a significant role in nitrogen-fixation in light. The most active nitrogen-fixation by microorganisms occurred in the soil shortly after it had been submerged under light. The longer the submergence, the less nitrogen microorganisms were fixed. In a greenhouse experiment, the fixed nitrogen appeared not to be immediately available to the rice plant. The amount of nitrogen that can be fixed in the field by nitrogen-fixing microorganisms in paddy water was estimated using the acetylene reduction method during the rice-growing period. The amount of nitrogen fixation by these microorganisms is not sufficient to account for the amount of nitrogen uptake by rice during the rice-growing period.     

秸秆还田方式对根际固氮菌群落及花生产量的影响

  【目的】  固氮微生物是土壤中重要的功能微生物,其多样性和群落组成变化能够影响土壤氮素固定与氮循环过程,探究不同秸秆还田方式对根际土壤固氮菌多样性和群落组成的影响机制具有重要意义。  【方法】  基于中国科学院鹰潭红壤生态实验站花生单作系统不同秸秆还田长期定位试验,设置不施肥对照(CK)、单施化肥(NPK)、NPK肥+秸秆还田(NPKS)、NPK肥+秸秆猪粪配施(NPKSM)和NPK肥+秸秆生物炭(NPKB) 5个处理,利用高通量测序技术,分析不同秸秆还田方式下根际固氮菌多样性和群落组成的变化特征。  【结果】  秸秆还田处理下土壤有机碳(SOC)、速效钾、全磷、有效磷、全氮含量提升,其中以NPK肥+秸秆猪粪配施(NPKSM)处理效果最佳。秸秆还田增加了固氮微生物多样性,并显著改变其群落组成,在纲水平上固氮菌以α-变形菌纲(Alphaproteobacteria,82.5%)为优势类群;在属水平上以慢生根瘤菌属(Bradyrhizobium,51.9%)为优势类群。土壤有效磷是影响固氮菌多样性指数的主要因素,而土壤pH、SOC、速效钾、全磷、有效磷、全氮和铵态氮是影响固氮菌群落组成的主要因素。结构等式方程研究结果表明土壤有效磷和全氮通过改变δ-变形菌纲(Deltaproteobacteria)的相对丰度和固氮菌群落组成间接影响花生产量。  【结论】  秸秆还田显著提升了土壤肥力,土壤有效磷是根际固氮菌多样性和群落组成改变、花生产量提高的重要驱动因素,通过提高Deltaproteobacteria的相对丰度促进了花生增产。本研究为建立合理的秸秆还田措施,增强生物固氮潜力以及提升红壤肥力与健康提供科学依据。     

受固氮活性制约的蓝藻固氮去铵阻抑及其与生理条件的关系

在不同气体环境中培养的蓝藻Ａｎａｂａｅｎａ７１２０固氮活性各异，Ａｒ＋ＣＯ２中最大，空气中次之，Ａｒ中最小。固氮活性高者去铵阻抑速率大，反之则小。它们对各种生理条件的反应不一样，固氮活性高者，其去铵阻抑速率受氧和氮的抑制小。在氢和氧加合条件下，三种活性的蓝藻去铵阻抑均加快，但活性低者慢而小些。光强减弱或加光合抑制剂时，固氮活性低者去铵阻抑速率显著小于活性高者，而添加外源的蔗糖或丙酮酸时，也是固氮活     

Effect of the introduction of the nitrogen-fixing bacteria <Emphasis Type="Italic">Pseudomonas putida</Emphasis> 23 on the nitrogen balance in soil

The inoculation of red beets with the nitrogen-fixing bacteria Pseudomonas putida 23 increased the activity of the nitrogen fixation in the rhizosphere of the plants grown on meadow soil in the central part of the Oka River floodplain. The yield of the red beets and the uptake by plants of nitrogen from the soil and from the ¹⁵N-labeled nitrogen fertilizer applied on the trial microplot increased significantly. A statistically significant additional fixation of nitrogen from the atmosphere and a positive balance of nitrogen in the soil-plant system without significant changes in the bulk content of the soil nitrogen after the plant growing were found in a greenhouse experiment with the application of P. putida. It can be supposed that the excessive nitrogen determined in this system is related to the incorporation into plants of atmospheric nitrogen fixed in the rhizosphere of the inoculated plants. The application of P. putida 23 makes it possible to decrease the rates of NPK fertilizer by two times without losses in the yield of red beets.     

固氮芽孢杆菌GD272的筛选鉴定及其固氮性能研究

对所选育出的一株固氮芽孢杆菌（编号为GD272）进行了形态、生理生化测定和接种效果研究。通过16S rDNA基因比对以及生理生化鉴定表明,该菌株属于芽孢杆菌Bacillus sp.。乙炔还原法测定显示该菌株具有较高的固氮酶活性;小白菜盆栽试验看出,接种GD272菌达到了施用化学氮肥的同等效果。菌株GD272在固氮微生物肥料生产中具有较好的开发应用前景。     

利用15N2直接标记法研究水稻种植对稻田固氮量和固氮活性的影响

稻田固氮对土壤维持肥力有着重要的作用,但水稻种植与固氮菌及其活性之间的关系尚不清楚。本试验利用~(15)N_2直接标记法测定了下位砂姜土发育的简育水耕人为土在种水稻和不种水稻条件下的生物固氮量,及其在土壤不同层次(0～1、1～5、5～15 cm)和水稻中的分配,并通过实时荧光定量PCR技术测定了土壤中固氮菌nifH DNA及RNA基因数量。结果表明:种水稻处理显著提高了土壤各层固氮量,尤其提高了1～5 cm和5～15 cm土层土壤固氮量对总固氮量的贡献;种水稻处理的总固氮量是不种水稻处理的10.3倍;水稻植株中生物固定的氮占总固氮量的31.48%;在0～1 cm土层,种水稻处理显著提高了nifH RNA基因数量,而对nifH DNA基因数量的增加不显著。可见,水稻种植没有增加固氮菌的数量,稻田固氮量的增加是因为水稻种植极大地促进了固氮菌nifH基因的表达,提高了固氮菌的固氮活性。     

Comparative toxicity of Al3+, Yb3+, and La3+ to root-tip cells differing in tolerance to high Al3+ in terms of ionic potentials of dehydrated trivalent cations

Abstract

Green manure legumes are often used to compare biomass production as well as nitrogen-fixing capacity. Mineral deficiency often limits the symbiotic nitrogen fixation of many legumes, thus limiting their productivity despite their high yielding potential (O’Hara et al. 1988; Flis et al. 1993). Leguminous species require large amounts of P for growth, nodulation, and nitrogen fixation. Consequently, they are often unable to grow in acid soils with low available P. The low P availability in tropical acid soils often arises from fixation of P by Al and Fe in soil. Generally, Al and Fe-phosphates are relatively unavailable to plants (McLachlan 1976; Ae et al. 1990).     

毛竹入侵森林对固氮微生物群落结构和丰度的影响

【目的】 毛竹入侵能够对生态系统的各个方面产生影响,包括森林生态系统氮循环。生物固氮是森林生态系统氮循环的重要一环,研究毛竹入侵过程中土壤固氮微生物的变化,可为毛竹入侵对生态功能影响的评价提供理论依据。 【方法】 选取天目山自然保护区的三个毛竹入侵带 (青龙山、石门洞和进山门),分别在三个入侵带中沿毛竹入侵方向采集毛竹纯林、毛竹与原林混交林以及原林地的表层土壤,分析pH、有机碳、碱解氮、有效磷和速效钾等土壤化学性质,应用基于nifH功能基因的末端限制性片段长度多态性 (T-RFLP) 和荧光定量PCR (qPCR) 技术,分析土壤固氮微生物群落结构和丰度的变化。 【结果】 毛竹入侵后土壤化学性质呈现三种不同情况,在三个入侵带中总体上升的有pH、有机碳、碱解氮、有效磷,下降的为速效钾,而硝态氮的变化均不显著,总体上,土壤养分含量在毛竹入侵后有所上升。土壤固氮微生物的丰度随毛竹入侵过程降低,其中进山门入侵带的变化显著 (P < 0.05)。土壤固氮微生物T-RFLP的结果显示,48 bp在三个入侵带中均为优势片段;不同的T-RFs在毛竹入侵过程中变化各异,有的片段在毛竹入侵后消失,有的片段在毛竹入侵后出现,有的片段变化不显著。土壤固氮微生物群落的Shannon指数和均匀度指数随毛竹入侵降低,Simpson指数相反,这些指标只在石门洞入侵带的不同林分之间存在显著差异 ( P < 0.05);多响应置换过程分析 (MRPP) 显示,石门洞和进山门入侵带毛竹入侵前后土壤固氮微生物群落结构有显著变化 ( P < 0.05);冗余分析 (RDA) 结果表明,土壤性质对固氮微生物群落变化的解释率普遍较低 (低于30%),三个入侵带显著影响土壤固氮微生物群落变化的土壤化学性质各不相同,并且三个入侵带的样点在RDA坐标图中分布格局并不统一。 【结论】 即使同一自然保护区的三个毛竹入侵带,土壤固氮微生物群落随着入侵过程的变化并不一致,母岩和原有植被产生的综合作用导致土壤固氮微生物群落发生变化,但需结合更多的因素进行进一步地探讨。      

山桃沙棘混交林养分状况研究

分析了半干旱退化山区荒山5 a生沙棘×山桃混交林与山桃纯林系统的土壤养分状况。结果发现,山桃×沙棘混交林土壤有机质和氮素有明显的提高。在林木根系活动层40—60 cm范围土壤有机质、氮素增量高于山桃纯林,土壤速效磷增幅较小,土壤速效钾有少量增加。     

水稻根际固氮量及根系不同部位的固氮活性

采用乙炔还原法测定水稻根际原状土粒的固氮活性,以3∶1的换算比例推算为田间固氮量。每季水稻根际固氮量早稻为0.4-0.45斤氮/亩/66天;晚稻为0.6-0.79斤氮/亩/72天。其中早稻80—90%的氮素在抽穗至成熟期固定,而晚稻在这段期间固定的氮素占总数的40—52%。经多次测定表明:水稻根系不同部位固氮活性的趋势是,埋入土中带根基茎段2.5厘米活性最高,根基以下3厘米根段次之,3厘米以下根段活性很低,水稻根系结合的腐生性细菌以固氮菌占优势,同时也含有少量的丁酸固氮梭菌。     

钼对甘蔗体内固氮菌的固氮酶活性的影响

以巴西固氮甘蔗品种B1和B8为材料,在温室桶栽砂培条件下,对甘蔗施以含不同钼水平的营养液,以了解钼对甘蔗体内固氮菌的固氮酶活性的效应。结果表明,在含氮条件下,较低浓度的钼处理能提高甘蔗根内固氮菌的固氮酶活性,高浓度钼处理则能提高B1茎、叶片和B8茎中固氮菌的固氮酶活性;在无氮条件下,钼处理提高了B1叶片和茎中固氮菌的固氮酶活性,而B1根及B8根、茎、叶中固氮菌的固氮酶活性有所降低。此外,甘蔗根、茎、叶中固氮菌的固氮酶活性之间关系较为密切,根中固氮菌的固氮酶活性与叶片和茎中的都呈负相关。上述结果说明:巴西甘蔗在本地也具有一定的固氮能力;在不缺氮条件下,钼处理较利于调节甘蔗体内固氮菌的固氮酶活性,促进其固氮;而在缺氮条件下,钼处理不利于调节甘蔗体内固氮菌的固氮酶活性,抑制其固氮作用。     

The effect of acidity on the distribution and symbiotic efficiency of rhizobia in Lithuanian soils

The distribution and symbiotic efficiency of nodule bacteria Rhizobium leguminosarum_bv. trifolii F., Sinorhizobium meliloti D., Rhizobium galegae L., and Rhizobium leguminosarum bv. viciae F. in Lithuanian soils as dependent on the soil acidity were studied in the long-term field, pot, and laboratory experiments. The critical and optimal pH values controlling the distribution of rhizobia and the symbiotic nitrogen fixation were determined for every bacterial species. The relationship was found between the soil pH and the nitrogen-fixing capacity of rhizobia. A positive effect of liming of acid soils in combination with inoculation of legumes on the efficiency of symbiotic nitrogen fixation was demonstrated.     

不同耕作方式下玉米农田土壤养分及土壤微生物活性变化

通过连续3年的野外调查与室内分析试验,研究了不同耕作方式（翻耕、旋耕、免耕）对玉米农田土壤养分及土壤微生物活性的影响。结果表明,不同耕作方式下土壤pH值略显酸性,土壤容重与土壤总孔隙度变化趋势相反;土壤容重基本表现为:免耕 > 翻耕 > 旋耕;土壤总孔隙度TSP基本表现为:翻耕 > 旋耕 > 免耕,不同耕作方式差异均显著（p<0.05）。不同耕作方式下土壤养分（有机碳、全氮含量）和有效养分（有效磷、铵态氮和硝态氮）均呈现出一致性规律,大致表现为翻耕 > 旋耕 > 免耕,不同耕作方式下土壤全磷含量差异均不显著（p > 0.05）;与免耕相比,土壤微生物量碳和氮、土壤微生物数量（细菌、真菌、放线菌、固氮菌和纤维素菌）均有明显的增加,大致表现为:翻耕 > 旋耕 > 免耕。土壤微生物活度的变化范围为0.38~0.69,依次表现为翻耕 > 旋耕 > 免耕,不同耕作方式下土壤微生物活度差异均显著（p<0.05）。不同耕作方式下土壤微生物量碳周转率高于氮周转率,说明微生物量碳更新比微生物量氮快,其中翻耕处理下土壤微生物量碳和氮更新较旋耕和免耕快。通径分析发现,不同土壤环境因子对土壤微生物活度产生直接和间接负作用,其中有机碳、全氮、硝态氮、铵态氮、细菌数量对土壤微生物活度产生直接效应;土壤微生物量碳、微生物量氮、真菌数量和固氮菌数量对土壤微生物活度产生间接效应。     

阔叶林改种毛竹(Phyllostachys pubescens)后土壤固氮细菌 nifH 基因多样性的变化

【目的】毛竹是喜氮植物,土壤氮素水平对毛竹生长至关重要。生物固氮是土壤氮素的重要来源,因此,探索阔叶林改种毛竹后土壤固氮细菌和土壤氮素的变化具有重要意义。【方法】选择立地条件相近的毛竹林(100多年前由阔叶林改种而来)和阔叶林,每种林地在东北坡向位置随机选择4个10 m×10 m标准样地,每个标准样地选取5个采样点,分层采集0—20 cm(表层)和20—40 cm(次表层)土壤样品,分析了土壤pH、有机碳、碱解氮、有效磷、速效钾和含水量等常规理化性质; 采用引物对AQER和PolF,以土壤总DNA为模板扩增了固氮细菌功能基因(nifH )片段,应用变性梯度凝胶电泳(DGGE)和实时荧光定量PCR(Real-time PCR),分析了固氮细菌群落结构、多样性以及丰度(nifH 基因拷贝数)变化; 通过基因克隆测序对土壤固氮细菌进行初步鉴定。【结果】阔叶林改种毛竹后土壤pH显著(P0.05)提高; 毛竹林土壤的含水量、碱解氮以及表层土壤的速效钾显著高于(P0.05)同层的阔叶林土壤,而有效磷则显著(P0.05)低于同层的阔叶林土壤。总体来说,阔叶林改种毛竹后土壤养分含量明显提高; 阔叶林土壤固氮细菌DGGE条带数以及多样性指数(Shannon-Wiener index)都高于毛竹林; 基于DGGE条带信息的聚类分析和主成分分析(PCA)结果表明,阔叶林和毛竹林区分为2个类群,而同一林分的不同土层之间差异较小; 实时荧光定量PCR结果显示,毛竹林土壤的固氮细菌 nifH 基因丰度显著(P0.05)高于阔叶林土壤; 通过克隆测序,14个阳性克隆分别属于2个不同的菌属,其中13个均为Bradyrhizobium,1个为Azohydromonas lata,条带序列与已知序列的相似度为93%~98%。【结论】阔叶林改种毛竹后土壤固氮细菌的种类减少,而功能基因丰度却明显增加; 土壤氮素水平明显提高,这可能是土壤固氮能力增强的结果。     

Increase in the Nitrogen Content of Soil by the Introduction of Earthworms into Soil

The contribution of an earthworm species ( Amynthas vittatus ) to the increase of the nitrogen content of soil was examined. Three specimens of adult earthworms were introduced into 300 g of soil (Gray Lowland soil, silty clay) supplemented with 1% carboxymethyl cellulose in a container and incubated for 32 d at 22°C in the dark. The contents of total-N, NH₄-N and NO₃-N, and the population of aerobic nitrogen-fixing bacteria in soil significantly increased after incubation with the earthworms, while the natural abundance of ¹⁵N (δ¹⁵N) in soil decreased. The amount of nitrogen in the earthworms did not decrease during the incubation in the microcosm. Both acetylene reduction activity of the microcosm and incorporation of ¹⁵N to soil from atmospheric ¹⁵N₂ were significantly enhanced by the introduction of the earthworms into soil, though the observed increment of nitrogen in soil was much higher than the estimated one based on the nitrogen-fixing activity. The results obtained in the present study indicated that the earthworms increased the nitrogen content of soil, presumably due to the enhancement of the nitrogen-fixing activity of the soil from the microcosm by the earthworms.     

施氮量对玉米/大豆套作系统土壤微生物数量及土壤酶活性的影响

为揭示玉米/大豆套作体系下土壤氮素转换的调控机理和根际微生态效应,以种植模式为主因素[设玉米单作(MM)、大豆单作(SS)和玉米/大豆套作(IMS)3种处理],以玉米、大豆施氮总量(玉米、大豆施氮比例为3∶1)为副因素[设不施氮(NN,0 kg?hm~(-2))、减量施氮(RN,180 kg?hm~(-2))和常量施氮(CN,240 kg?hm~(-2))3个处理],研究了玉米/大豆套作系统下不同施氮量对作物根际土壤微生物数量及土壤酶活性的影响。结果表明:与相应单作相比,套作下玉米根际土壤真菌、放线菌数量分别提高25.37%和8.79%;套作大豆根际土壤真菌、放线菌、固氮菌数量高于单作大豆;套作玉米根际土壤蛋白酶、脲酶活性和套作大豆根际土壤蛋白酶活性均显著升高。各施氮水平间,减量施氮下玉米、大豆根际土壤真菌数量较常量施氮和不施氮均有所提高;施氮提高了玉米、大豆根际土壤放线菌数量;大豆根际土壤固氮菌数量以减量施氮最高,比不施氮和常量施氮高17.78%和5.67%;玉米根际土壤蛋白酶活性、脲酶活性和大豆根际土壤脲酶活性均以减量施氮为最高。适宜的施氮量不仅能增加玉米/大豆套作土壤中真菌、放线菌、固氮菌的数量,还能提高土壤蛋白酶、脲酶活性,调节土壤氮素的转化,促进玉米/大豆对土壤中氮素的吸收,实现节能增效。