Liquid extraction of low molecular mass organic acids and hydroxamate siderophores from boreal forest soil

Low molecular mass organic acids (LMMOAs) and hydroxamate siderophores (HS) are molecules secreted by microbes and have previously been found in soil solution and in cultures. Mycorrhizal fungi are suggested to be involved in the nutrient uptake processes of trees and weathering of minerals. In this study soil samples taken from the O and E horizons of a podzol were extracted with 10 mM potassium phosphate buffer at pH 7.2. Variable parameters included addition of methanol to the extraction buffer and the use of ultrasonication or rotary shaking during extraction. LMMOAs and HS content of the soil extracts were determined. Analysis of soil extracts were carried out by liquid chromatography mass spectrometry (LC–MS) and the extraction results compared to results for soil solution samples obtained by centrifugation of the soils sampled. The extraction yields were significantly increased by addition of methanol to the extraction buffer, especially for the O horizon samples. Rotary shaking of the samples for 90 min gave slightly higher yields than ultrasonication for 15 min but the reduction in extraction time makes ultrasonication an attractive option. Of the HSs determined, ferricrocin was found in all samples. Optimal extraction conditions showed citric acid and isocitric acid to be the most abundant organic acids in the O and E horizons, respectively.     

红壤中低分子量有机酸的吸附动力学

低分子量有机酸在土壤中广泛存在尤其是在根际土壤中,其主要来源于动植物残体的分解、微生物的代谢、植物根系的分泌和土壤中有机物的转化等。由于羧基的强络合作用,低分子量有机酸一旦进入土壤将很快被土壤吸附,影响土壤的表面电荷性质和动电性质,进而影响其在土壤中的作用。关于低     

低分子量有机酸对红壤磷酸单酯酶活性的影响

采用室内模拟试验研究了低分子量有机酸(柠檬酸、草酸、苹果酸、酒石酸)对红壤磷酸单酯酶活性的影响。结果表明,低浓度有机酸(<1μmol g-1)对磷酸单酯酶活性有促进作用,且促进作用大小依次为柠檬酸≈草酸>苹果酸>酒石酸;而高浓度有机酸(>5μmol g-1)则为抑制作用,且抑制作用大小依次为柠檬酸>草酸>苹果酸>酒石酸。当体系pH趋向酶促反应最佳pH时,磷酸单酯酶活性增强;反之,当体系pH远离酶促反应最佳pH时,磷酸单酯酶活性减弱。有机酸根一方面通过羧基的辅助作用提高磷酸单酯酶活性;另一方面通过释放土壤A l3+、Fe3+等金属离子,对土壤磷酸单酯酶活性有一定的抑制作用。     

温度和水土比对红壤吸附低分子量有机酸的影响

通过一次平衡法考察了温度和水土比对红壤吸附草酸、柠檬酸、酒石酸和苹果酸的影响。结果表明,有机酸的吸附量、最大吸附量（Sm）及吸附亲和力常数（K）均随温度的升高而增加,随着水土比的增加而降低。温度和水土比变化对有机酸吸附的影响程度因有机酸种类而异,影响程度大小与土壤对有机酸的吸附亲和力（K）大小顺序相反。45℃ 时,土壤对草酸、柠檬酸、酒石酸和苹果酸的吸附量分别是 25℃ 时的 1.19、1.22、1.24 和 1.28 倍;100:1 水土比条件下,草酸、柠檬酸、酒石酸和苹果酸的吸附量分别较 10:1 水土比条件下减少了 48.3%、54.9%、63.5% 和 76.1%。     

红壤主要土壤组分对低分子量有机酸吸附的研究

通过一次平衡法研究了双氧水去除有机质、添加1%腐殖酸和DCB法(连二亚硫酸钠-柠檬酸钠-碳酸氢钠)去除游离氧化铁、铝对红黏土发育的红壤吸附草酸、柠檬酸、酒石酸和苹果酸的影响。结果表明,去除有机质后,由于受溶液pH、表面吸附点位变化、土壤结构变化、表面基团活性变化、草酸根生成等多种因素的影响,红壤对低分子量有机酸的吸附量虽略有增加,但增加并不明显。添加腐殖酸培养一个月后,由于受土壤表面电荷变化、吸附点位覆盖、氧化铁活性改变、基团质子化等因素的影响,红壤对低分子量有机酸的吸附量虽有轻微的减少,但减少也不明显。去除占土壤总量3.79%的游离氧化铁、铝后,土壤表面正电荷将会显著减少,而导致红壤对低分子量有机酸的吸附量显著减少,其减少程度因有机酸种类而异,与原土对有机酸的最大吸附量(Sm)大小顺序相反。     

低分子量有机酸对红壤无机磷活化的作用

采用室内模拟试验研究了低分子量有机酸,如草酸、柠檬酸、酒石酸和苹果酸在红壤无机磷活化中的作用。结果表明,相同浓度下,有机酸活化土壤磷的能力为柠檬酸酒石酸苹果酸。低浓度(0.5mmolL-1)时,草酸活化能力最小;高浓度(≥5mmolL-1)时,其活化能力最大。对同一种有机酸而言,土壤各无机磷形态活化量均随pH的升高而降低;且在同一酸度下,其活化量以铝磷(Al-P)为最多,铁磷(Fe-P)和钙磷(Ca-P)次之,闭蓄态磷(O-P)则最少。有机酸活化土壤无机磷酸盐应该是质子酸效应和有机酸阴离子络合效应共同作用的结果,且与磷酸盐的溶度积常数密切相关。研究结果对根系土壤无机磷素循环研究有着重要的意义。     

pH、离子强度和介电常数对低分子量有机酸在红壤中吸附行为的影响

通过一次平衡法考察了pH、离子强度和溶剂介电常数对红壤吸附低分子量有机酸的影响。研究结果表明,随着溶液pH值的升高有机酸的吸附量降低,其中草酸和酒石酸的吸附量在pH 3.5 ~ 5.0范围内随着pH值的升高而急剧下降,之后缓慢下降（pH 5.0 ~ 7.0）。柠檬酸、酒石酸和苹果酸的吸附量在1 ~ 2 mmol/L初始浓度范围内随着离子强度的增加没有明显变化,但在2 ~ 20 mmol/L初始浓度范围内却随着离子强度的增加而增加。红壤对草酸、柠檬酸、酒石酸和苹果酸的吸附量均随着溶剂介电常数的减小而增加。当溶剂中含6% 的乙醇时,草酸、柠檬酸、酒石酸和苹果酸的吸附量分别是对照（不含乙醇）的1.05、1.05、1.11和1.31倍。     

Fate of low molecular weight organic substances in an arable soil: From microbial uptake to utilisation and stabilisation

Microbial uptake and utilisation are the main transformation pathways of low molecular weight organic substances (LMWOS) in soil, but details on transformations are strongly limited. As various LMWOS classes enter biochemical cycles at different steps, we hypothesize that the percentage of their carbon (C) incorporation into microbial biomass and consequently stabilisation in soil are different.Representatives of the three main groups of LMWOS: amino acids (alanine, glutamate), sugars (glucose, ribose) and carboxylic acids (acetate, palmitate) – were applied at naturally-occurring concentrations into a loamy arable Luvisol in a field experiment. Incorporation of ¹³C from these LMWOS into extractable microbial biomass (EMB) and into phospholipid fatty acids (PLFAs) was investigated 3 d and 10 d after application. The microbial utilisation of LMWOS for cell membrane construction was estimated by replacement of PLFA-C with ¹³C.35–80% of initially applied LMWOS-¹³C was still present in the composition of soil organic matter after 10 days of experiment, with 10–24% of ¹³C incorporation into EMB at day three and 1–15% at day 10. Maximal incorporation of ¹³C into EMB was observed from sugars and the least from amino acids. Strong differences in microbial utilisation between LMWOS were observed mainly at day 10. Thus, despite similar initial rapid uptake by microorganisms, further metabolism within microbial cells accounts for the specific fate of C from various LMWOS in soils.¹³C from each LMWOS was incorporated into each PLFA. This reflects the ubiquitous utilisation of all LMWOS by all functional microbial groups. The preferential incorporation of palmitate into PLFAs reflects its role as a direct precursor for fatty acids. Higher ¹³C incorporation from alanine and glucose into specific PLFAs compared to glutamate, ribose and acetate reflects the preferential use of glycolysis-derived substances in the fatty acids synthesis.Gram-negative bacteria (16:1ω7c and 18:1ω7c) were the most abundant and active in LMWOS utilisation. Their high activity corresponds to a high demand for anabolic products, e.g. to dominance of pentose-phosphate pathway, i.e. incorporation of ribose-C into PLFAs. The ¹³C incorporation from sugars and amino acids into filamentous microorganisms was lower than into all prokaryotic groups. However, for carboxylic acids, the incorporation was in the same range (0.1–0.2% of the applied carboxylic acid ¹³C) as that of gram-positive bacteria. This may reflect the dominance of fungi and other filamentous microorganisms for utilisation of acidic and complex organics.Thus, we showed that despite similar initial uptake, C from individual LMWOS follows deviating metabolic pathways which accounts for the individual fate of LMWOS-C over 10 days. Consequently, stabilisation of C in soil is mainly connected with its incorporation into microbial compounds of various stability and not with its initial microbial uptake.     

低分子量有机酸对红壤和黄褐土K+吸附动力学的影响

采用连续流动法研究了低分子量有机酸影响下供试土壤的钾素吸附动力学特征,探讨了描述土壤K 吸附动力学的最优模型。结果发现,有机酸（苹果酸、柠檬酸）作用下,红壤吸附的K 量均低于对照（不加有机酸）处理,0.1 mmol/L草酸处理除外。黄褐土对K 的吸附因K 浓度的不同而差异明显。当K 浓度为0.1 mmol/L时,有机酸作用下黄褐土吸附的K 量低于对照,而当K 浓度为1.0 mmol/L时,有机酸作用下的黄褐土K 吸附量高于对照。K 吸附反应速度与时间的自然对数lnt间存在良好的线性关系（R0.05=0.754,R0.01=0.874）。1.0 mmol/L KCl处理的初始吸附速率较高,反应速度降低的较快。对红壤、黄褐土吸附K 的数据进行拟合,双常数方程、指数方程和Elovich方程拟合效果较好,都达到了极显著水平,一级动力学方程拟合效果不好。双常数方程的相关系数（R）高于Elovich方程和指数方程,双常数方程是描述红壤和黄褐土在有机酸作用下K 吸附动力学的最优模型。结果表明,有机酸对两种土壤吸附能力的影响均表现为草酸＞柠檬酸＞苹果酸。有机酸作用下红壤和黄褐土的K 吸附过程主要受土壤电荷的影响。     

低分子量有机酸类物质对红壤和黑土磷有效性的影响

在20℃恒温培养条件下,研究低分子量有机酸类的钠钙盐(LA)和低分子量有机酸类与混合氨基酸的钠钙盐(LAA)不同用量水平对红壤和黑土磷有效性的影响。研究表明,两种低分子量有机酸类混合物在各不同水平下均能显著提高红壤和黑土磷有效性;随着培养的进行,添加有LA、LAA的各处理对红壤和黑土中磷有效性均有明显提高。无论是LA或LAA,红壤中以半量处理对提高磷有效性作用效果更为明显,而黑土中则以LAA作用效果更好,且以常量LAA处理的效果最好,常量LA和LAA处理间差异显著。在添加磷肥的基础上,与不加有机酸物质处理(CKp)相比,添加常量LA、LAA和半量LA(0.5 LA)、LAA(0.5 LAA)处理,在红壤中磷固定率平均值分别降低10.5%、22.3%和11.3%、19.4%;在黑土中分别降低8.6%、10.6%和14.1%、11.8%。LA和LAA对降低红壤和黑土磷固定率效果显著,红壤以半量处理的效果较好,黑土则以常量LAA处理效果最好。     

The carbon we do not see—the impact of low molecular weight compounds on carbon dynamics and respiration in forest soils: a review

Dissolved organic matter (DOM), typically quantified as dissolved organic carbon (DOC), has been hypothesized to play many roles in pedogenesis and soil biogeochemical cycles, however, most research to date concerning forest soils has focussed on the high molecular weight (HMW) components of this DOM. This review aims to assess the role of low molecular weight (LMW) DOM compounds in the C dynamics of temperate and boreal forest soils focussing in particular on organic acids, amino acids and sugars. The current knowledge of concentrations, mineralization kinetics and production rates and sources in soil are summarised. We conclude that although these LMW compounds are typically maintained at very low concentrations in the soil solution (<50 μM), the flux through this pool is extremely rapid (mean residence time 1-10 h) due to continued microbial removal. Due to this rapid flux through the soil solution pool and mineralization to CO₂, we calculate that the turnover of these LMW compounds may contribute substantially to the total CO₂ efflux from the soil. Moreover, the production rates of these soluble transitory compounds could exceed HMW DOM production. The possible impact of climate change on the behaviour of LMW compounds in soil is also discussed.     

Low molecular weight organic acid adsorption in forest soils: effects on soil solution concentrations and biodegradation rates

Low molecular weight (LMW) organic acids are believed to play a key role in many rhizosphere and pedogenic processes; However, their efficiency is likely to depend on their susceptibility to sorption and biodegradation. The sorption characteristics of three organic acids (citrate, oxalate and acetate) and phosphate were examined over the concentration range 0-1000 μM in three coniferous forest soil profiles. Sorption to the soil's solid phase could be adequately described by the Langmuir equation with sorption capacity following the horizon series: B>C>E>O. The strength of anion sorption followed the series: phosphate>oxalate≥citrate?acetate. Calculations indicated that between 50 and 95% (O and E horizons) and >93% (B horizons) of these LMW organic acids entering the soil will become sorbed to the solid phase. The amount of organic acids predicted to be present on the solid phase at typical soil solution concentrations ranged from <1 to 1100 nmol g⁻¹ yielding adsorbed-to-solution ratios (adsorption coefficients) of between <0.1 and 3100. In the case of citrate, sorption to the solid phase significantly reduced its biodegradation potential by 35-99% depending upon the degree and type of sorption surface. The findings of this work are discussed in the context of the quantitative effects of adsorption on organic acids, their ecological functions and role in soil forming processes.     

气候因子对森林土壤有机碳影响的幅度效应研究

揭示不同幅度上气候因子对土壤有机碳(Soil organic carbon,SOC)影响的主控性变化,是预测未来气候变化对SOC演变趋势影响的基础。本文利用中国西南地区363个森林土壤剖面数据,基于大区、省和地级市3个幅度,研究了气候因子对森林SOC密度的影响随幅度变化的规律及不同幅度下的主控气候因子。结果表明,年均降水量与SOC密度的相关性均随着幅度的减小而减弱,而年均气温与SOC密度的相关性随幅度变化的规律不明显,有较强的区域差异。大区幅度上,SOC密度主要受年均降水量和年均气温的综合作用。省级幅度上,西藏自治区东部主控因子为年均降水量,而四川和云南两省为年均气温。地级市幅度上,各市的主控因子基本与其所属的省一致。气候因子对SOC密度变异的解释能力在大区幅度上约20%,且随着幅度的减小解释能力也逐渐减小。     

有机碳质量分数对森林土壤水分入渗过程的影响及其模拟

为了探讨三峡库区不同森林植物群落土壤有机碳质量分数与土壤水分入渗的关系,将森林土壤有机碳质量分数设定为独立变量,引入土壤有机碳质量分数对Horton土壤水分入渗模型进行修正。结果表明:1)不同森林植被群落类型土壤有机碳质量分数不同,表现为阔叶混交林>针阔混交林>针叶混交林>乔灌混交林;2)初渗速率、初渗速率和稳渗速率的差分别与土壤下层土壤有机碳质量分数呈线性相关关系。研究还发现,Horton模型经引入土壤有机碳指标后,新模型的入渗速率与入渗量模拟值与实测值相关系数明显提高,模拟结果更逼近实测值。研究结果为同类地区森林土壤水分入渗过程预测与模拟提供了一种新方法。     

浙江南部亚热带森林土壤植硅体碳的研究

植硅体封存有机碳(Phytolith-occluded organic carbon,Phyt OC)是一种稳定的有机碳形态。它由植物自身硅化作用产生,在植物死亡或凋落后归还于土壤,从而影响森林生态系统稳定性碳库的储量。本文以浙江庆元县5种不同亚热带典型森林立地土壤为研究对象,利用不同土层深度(0~10 cm、10~30 cm、30~60 cm和60~100 cm)土壤样品,分析土壤植硅体含量和植硅体碳含量,并估算土壤中植硅体碳储量。结果表明,毛竹林、杉木林、针阔混交林、阔叶林和马尾松林土壤植硅体含量(土壤剖面平均值)变化范围在8.14~19.74 g kg-1,其中毛竹林土壤植硅体含量最高。而植硅体中Phyt OC平均含量最高的为马尾松林(24.31 g kg-1),最低的为针阔混交林(13.06 g kg-1)。土壤Phyt OC/TOC比值随土层深度增加而急剧增加。统计分析表明,不同林分下土壤硅含量与土壤植硅体含量呈极显著相关关系(p0.01),与土壤Phyt OC含量之间呈显著的正相关关系(p0.05)。我国亚热带毛竹林、杉木林、马尾松林、阔叶林和针阔混交林1 m土体Phyt OC总储量分别为1.988×107、4.025×107、2.575×107、2.542×107和0.340×107 t。     

Determination of aluminium complexes of low molecular organic acids in soil solution from forest soils using ultrafiltration

Ultrafiltration (<1000 D) was evaluated as an analytical method for determination of Al bound to low molecular weight organic acids (LMWOA) in soil solutions from podzolised forest soils. The results were compared to those obtained by two chemical equilibrium models and a size exclusion chromatography method. The percentage of Al bound to LMWOAs was highest in the O horizon solutions (15–44%) and decreased in the deeper horizons. Citric acid was found to be the most important complex former. Generally the ultrafiltration method and modelling showed acceptable agreement. The method showed acceptable precision and good recovery of Al in the samples.     

植茶年限对土壤pH值、有机质与酚酸含量的影响

土壤pH值和有机质是土壤肥力的重要指标,土壤中的酚酸类物质也是一类重要的有机物质,与pH值和有机质关系复杂。本文以四川省名山区茶园土壤为对象,研究植茶年限对土壤pH值、有机质和酚酸含量的影响,并探讨它们之间的相互关系。结果显示:随着植茶年限的延长,各土层土壤的pH值均呈现不同程度的下降趋势,即植茶10年植茶7年植茶5年植茶3年水稻土,表层土壤酸化明显;随着植茶年限的延长,土壤有机质、总酚和水溶性酚含量均呈增加趋势。相关分析表明,0~10 cm土层土壤中,总酚与土壤pH值呈负相关;10~20和30~40 cm土层土壤中,水溶性酚与土壤pH值呈负相关;10~20 cm土层土壤中,总酚与土壤有机质呈正相关;在10~20、20~30和30~40 cm土层土壤中,水溶性酚与土壤有机质呈正相关。说明植茶土壤酚酸含量的增加是影响土壤pH值下降和有机质含量升高的重要因素。     

有机物料对白土土壤胡敏酸结构特征的影响

【目的】研究有机物料施入对白土土壤的腐殖质含量组成和胡敏酸(HA)结构特征的影响,为明确不同腐殖质组分对土壤肥力的影响提供理论依据。【方法】供试土壤为江苏省溧阳市南渡镇"白土改良大田示范试验核心区"的南方中低产水稻土(白土)。试验设秸秆还田(ST)、施有机肥(OM)和对照(CK,不施有机物)3个处理,培肥3年。同时采集试验田周围相邻的江苏省耕地质量监测点(2007~2013年)的每年施化肥(LAF)和长期不施肥(NF)的两种处理土壤进行比对研究。分别测定土壤的基本理化性质及其腐殖质含量的组成,并提取土壤胡敏酸(HA)固体样品利用红外光谱和元素分析来进行结构表征。【结果】秸秆还田和施有机肥处理的有机碳、全氮含量明显高于对照;与对照相比,施有机物料土壤HA的E4/E6比值增加,且秸秆还田施有机肥对照。红外光谱显示,试验区域和耕地监测点的不同处理土壤HA均在1650 cm-1处(酰胺I带)和1550 cm-1处(1500~1580 cm-1酰胺II带伸缩振动)有特征吸收。施有机肥和秸秆还田处理土壤HA的2920/1720、2920/1650比值显著大于对照。在元素组成上,OM、ST处理的土壤腐殖质(HA)中C、H、N的含量比均高于CK,相对长期施化肥(LAF)和不施肥(NF)的土壤有明显提高,而氧元素的含量呈降低的趋势;OM和ST处理土壤HA的[H]/[C]和[O]/[C]原子数比均低于CK;与LAF和NF处理相比,试验区域各处理土壤腐殖质的[H]/[C]和[O]/[C]原子数比均有明显降低。【结论】有机物料施入土壤后可增加土壤有机碳含量,改善土壤理化性质,提高作物产量和品质,且施入土壤的有机物料可转化为新的腐殖质,降低土壤的腐殖化程度。土壤腐殖质(HA)的红外光谱分析说明,白土土壤HA具有明显的酰胺类化合物特征。有机物料施入后使得土壤脂族性增强,羧基量减少,芳香度降低;秸秆还田和施有机肥处理与对照相比,土壤HA的[H]/[C]和[O]/[C]比均有下降的趋势,且HA的氮素含量明显增加,这显示有机物料施入后白土土壤腐殖质发生"脱水"过程,同时也反映了白土土壤腐殖质形成的特征。     

Near-infrared spectroscopy for analysis of chemical and microbiological properties of forest soil organic horizons in a heavy-metal-polluted area

In industrial areas, heavy metals may accumulate in forest soil organic horizons, affecting soil microorganisms and causing changes in the chemical composition of the accumulated organic matter. The objectives of this study were to test the ability of near-infrared spectroscopy (NIRS) to detect heavy metal effects on the chemical composition of forest soil O horizons and to test whether NIRS may be used to quantitatively determine total and exchangeable concentrations of Zn and Pb (Zn_t, Pb_t, Zn_ex, Pb_ex) and other chemical and microbial properties in forest soil O horizons polluted with heavy metals. The samples of O horizons (n = 79) were analyzed for organic C (C_org), total N and S (N_t, S_t), Zn_t, Pb_t, Zn_ex, Pb_ex, basal respiration (BR), microbial biomass (C_mic) and C_mic-to-C_org ratio. Spectra of the samples were recorded in the Vis-NIR range (400–2,500 nm). To detect heavy-metal-induced changes in the chemical composition of O horizons principal components (PC1–PC7) based on the spectral data were regressed against Zn_t + Pb_t values. A modified partial least squares method was used to develop calibration models for prediction of various chemical and microbial properties of the samples from their spectra. Regression analysis revealed a significant relationship between PC3 and PC5 (r = −0.27 and −0.34, respectively) and Zn_t + Pb_t values, indicating an effect of heavy metal pollution on the spectral properties of the O horizons and thus on their chemical composition. For quantitative estimations, the best calibration model was obtained for C_org-to-N_t ratio (r = 0.98). The models for C_org, N_t, and microbial properties were satisfactory but less accurate. NIRS failed to accurately predict S_t, C_org-to-S_t, Zn_t, Pb_t, Zn_ex, and Pb_ex.     

有机物质阴阳离子对调节土壤pH的作用

The process of organic materials increasing soil pH has not yet been fully understood. This study examined the role of cations and organic anions in regulating soil pH using organic compounds. Calcareous soil, acid soil, and paddy soil were incubated with different simple organic compounds, pH was determined periodically and CO2 emission was also measured. Mixing organic acids with the soil caused an instant decrease of soil pH. The magnitude of pH decrease depended on the initial soil acidity and dissociation degree of the acids. Decomposition of organic acids could only recover the soil pH to about its original level. Mixing organic salts with soil caused an instant increase of soil pH. Decomposition of organic salts of sodium resulted in a steady increase of soil pH, with final soil pH being about 2.7-3.2 pH units over the control. Organic salts with the same anions （citrate） but different cations led to different magnitudes of pH increase, while those having the same cations but different anions led to very similar pH increases. Organic salts of sodium and sodium carbonate caused very similar pH increases of soil when they were added to the acid soil at equimolar concentrations of Na^＋. The results suggested that cations played a central role in regulating soil pH. Decarboxylation might only consume a limited number of protons. Conversion of organic salts into inorganic salts （carbonate） was possibly responsible for pH increase during their decomposition, suggesting that only those plant residues containing high excess base cations could actually increase soil pH.