生物炭及炭基肥对土壤微生物群落结构的影响

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本站小编 Free考研考试/2021-12-26

0 引言

【研究意义】土壤微生物是土壤中物质循环和能量流动不可或缺的参与者^[1],也是土壤养分的“源”和“汇”,支撑着土壤肥力,对环境变化极为敏感,土壤中的微小变动都会引起其活性的变化^[2]。生物炭（Biochar）是由农林废弃物等在缺氧环境下加热裂解制成的一种稳定有机物,可作为富碳土壤改良剂^[3]。生物炭基肥料（炭基肥）是由生物炭与普通化学肥料复合而成的缓释肥料^[3,4,5]。磷脂脂肪酸（phospholipid fatty acid,PLFA）标记方法快速、准确、灵敏,技术成熟,可以有效测定土壤不同微生物群落生物量^[6,7]。通过研究生物炭和炭基肥定位施用后土壤微生物群落结构的变化来判断不同有机物料改良土壤效果,可以为农田土壤培肥提供生态学方面的理论依据。【前人研究进展】生物炭在制备过程中形成了多孔隙、大比表面积的结构,且具有高含碳量、多芳香烃、多极性官能团的组成^[3],理化性质稳定^[3,8],使其成为优良的土壤改良剂^[9]。相关研究^{[4, 10-11]}表明,生物炭施入土壤后提高了土壤含水量、pH、孔隙度、CEC以及养分水平,基于其本身的结构和性质,也能为微生物提供适宜生存的微环境,对不同微生物群落产生影响^[12]。细菌能够吸附到生物炭的表面而不易被淋洗^[13],且生物炭的吸附作用取决于其孔隙的大小^[14]。生物炭的微孔结构能为真菌提供“避难”场所,可以避免物种内的竞争^[15]。生物炭疏松多孔的结构特征,可如海绵一样保持与空气和水分的共通性,RONDON发现经生物炭处理的退化土壤固氮细菌能有效吸收来自大气的氮素^[16],原因是通过提高土壤通气性,增大氧分压来刺激氮细菌的活性^[17]。生物炭本身富含多种养分,特别是易被微生物利用的碳源和氮源,有利于微生物尤其是细菌的活动^[18]。O'NEILL利用同位素示踪法标记氨基酸和核苷酸,检测到添加生物炭后细菌活性和土壤呼吸速率增强,说明施用生物炭能够提高细菌对土壤碳的利用率^[19]。研究还发现,生物炭的施用量^[3],也会对土壤微生物的群落多样性产生影响。【本研究切入点】沈阳农业大学植物营养与施肥研究室于2009年春天布置生物炭及炭基肥棕壤改土定位试验,观测长期施用条件下的培肥效果,同时以传统的培肥改土措施——玉米秸秆还田和猪厩肥为对照,对比研究不同有机物料连续多年施用培肥改土的特征及优势。本试验的前期研究结果表明,生物炭和炭基肥等各有机物料处理均改善了土壤理化性质^[4],增加了土壤总有机碳和活性有机碳各组分碳含量,对土壤酶活性产生了一定影响^[21]。土壤有机碳和土壤酶的变化与土壤微生物的群落活性息息相关^[1],同时土壤微生物活性的变化与土壤养分的变异有关^[20],因此生物炭和炭基肥长期施用是否改变了土壤微生物群落值得探索。【拟解决的关键问题】本研究探讨生物炭和炭基肥连续施用7年后的土壤理化性质变化和土壤微生物群落结构特征及二者的相关关系,以期揭示生物炭培肥改土的生物学机理。

1 材料与方法

1.1 试验时间、地点和材料

田间微区试验于2009年5月开始,布置在沈阳农业大学国家花生产业技术体系土壤肥料长期定位试验基地（41°83′N,123°56′E）。试验区属温带湿润-半湿润季风气候,年均温度7.0—8.1℃,无霜期为147—164 d,年均降雨量574—684 mm。供试土壤属第四纪黄土母质发育的简育湿润淋溶土典型棕壤。2008年布置试验前土壤基本理化性质如下：土壤有机质1.53 g·kg^-1,土壤全氮0.86 g·kg^-1,土壤全磷0.42 g·kg^-1,土壤全钾20.40 g·kg^-1,碱解氮51.30 mg·kg^-1,有效磷4.60 mg·kg^-1,速效钾114.50 mg·kg^-1,pH 5.5（水土比5﹕1）。土壤以2﹕1型黏粒矿物为主,主要是水云母（Hydrous mica）,1﹕1型高岭石（Kaolinte）含量次之,蒙脱石（Montmorillonite）含量最低,CEC为8.32 cmol·kg^-1。土壤质地：砂粒含量510 g·kg^-1,粉粒含量280 g·kg^-1,黏粒（<2 μm）含量210 g·kg^-1,土质较黏。供试作物为花生,品种为阜花12。

1.2 试验设计

试验处理之间为相互对照,分为传统有机物料（玉米秸秆和猪厩肥）培肥改土处理和新型有机物料（生物炭颗粒和炭基肥）培肥改土处理。处理1：玉米秸秆（CS）4 500 kg·hm^-2 + NPK;处理2：猪厩肥（PMC）4 000 kg·hm^-2 + NPK;处理3：炭基肥（BF）750 kg·hm^-2（养分含量11-11-13）和处理4：生物炭颗粒（BIO）1 500 kg·hm^-2 + NPK。设计处理1—4等养分（全量NPK）,由于秸秆（CS）、猪厩肥（PMC）和生物炭颗粒（BIO）等有机物料本身养分含量低,以化肥（肥种类为尿素、过磷酸钙和硫酸钾）配施,使所有处理全量NPK养分相等,与炭基肥（BF）处理施用养分含量一致。表1为2015年各有机物料养分含量;生物炭颗粒为玉米芯450℃裂解,过筛添加胶结剂造粒,出炭率3﹕1;秸秆粉碎至2—3 cm,猪厩肥当年腐熟。每年5月初春播前将全部肥料当基肥施用,与土壤耕层混合后备垄,9月底收获后移除全部植株。试验设3次重复,随机区组排列,微区面积2 m²,微区间20 cm水泥埂分隔,微区内花生采用大垄双行种植,每池种植30穴。
Table 1
表1
表1各有机物料干基养分（%）和pH
Table 1The dry-based nutrient contents (%) and pH of different organic resources

有机物料 Organic resources	C	N	P₂O₅	K₂O	pH值
玉米秸秆CS	44.44	1.01	0.21	0.93	——
猪厩肥PMC	29.20	1.44	1.08	0.94	7.21
炭基肥BF	7.73	11.00	13.00	13.00	9.14
生物炭颗粒BIO	33.32	0.50	0.84	0.59	7.06

有机物料的养分含量为2015年的测定值 The nutrient of resources was measured in 2015CS: Corn straw-returning; PMC: Piggery manure compost; BF: Biochar-based compound fertilizer; BIO: Biochar. 下同The same as below

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1.3 试验方法

1.3.1 采集处理土样 2015年花生收获期（9月28日）采集微区0—20 cm土层土样,混合均匀后分两份,一份风干后用于土壤基本理化性质测定。另一份放入-80℃冰箱冷藏保存,用于冷冻干燥提取活体微生细胞膜脂肪酸。
1.3.2 基本理化性质测定土壤含水量（MC）测定
采用恒温箱烘干法;土壤全氮（TN）测定采用Dumas干烧法（德国VarioEL-Ⅲ型元素分析仪）;土壤有机碳（SOC）测定采用重铬酸钾容量法;土壤全钾（TK）测定采用氢氧化钠熔融-火焰光度计比色法（Sherwood M410火焰光度计）;pH采用电极法按土水比1﹕5测定（雷磁PHS-3G）。
1.3.3 PLFA提取与测定
（1）PLFA的提取采用BOSSIO等^[22]的方法,测定采用MIDI公司MIS系统鉴定。具体分为以下几步。
①浸提称取4.000 g冻干土样于聚四氟乙烯离心管中,加入15.2 mL混合浸提液（加液顺序依次为：3.2 mL柠檬酸缓冲液,4.0 mL氯仿,8.0 mL甲醇）。25℃避光振荡2.5 h,4 000 r/min离心10 min后上清液转移到提前加入8.0 mL氯仿和8.0 mL柠檬酸缓冲液的玻璃试管中,重复上述步骤一次并转移上清液,最终使氯仿﹕甲醇﹕柠檬酸缓冲液体积比=1﹕1﹕0.9。振荡试管1 min并定时放气。提取液避光静置过夜待分层。
②分离用玻璃吸管吸取分层后的上层水溶性液体（一样一只吸管,不要吸到下层氯仿）。试管于通风橱内避光条件通入纯N₂吹干氯仿层（保证整个过程试管充满N₂）。浓缩后的脂肪酸用5 mL氯仿分5次转移到氯仿活化的硅胶固相萃取柱内,依次用8 mL氯仿、16 mL丙酮和8 mL甲醇洗脱萃取柱,收集甲醇洗脱液于玻璃离心管内,通风橱避光条件下纯N₂吹干甲醇相。
③甲酯化玻璃离心管内加入1.0 mL甲醇-甲苯混合溶液（体积比1﹕1）和1.0 mL的0.2 mol·L^-1KOH-甲醇溶液,混匀后35℃水浴15 min。冷却至室温后依次加入2.0 mL去离子水、0.3 mL的1 mol·L^-1HAc和2.0 mL正己烷,涡旋30 s后2 500 r/min离心10 min,使用玻璃吸管吸取上层正己烷相于样品瓶内（一样一只吸管）,重复上述步骤一次合并正己烷,通风橱内避光条件下纯N₂吹干正己烷,即得到甲酯化磷脂脂肪酸（FAME）。
④PLFA图谱分析样品瓶内加入150 μL正己烷溶解样品,加入50 μL的40 μg·mL^-119:0甲基酯作内标用于定量,过气相色谱柱（Agilent GC 7890A）,利用美国MIDI公司开发的Sherlock Microbial Identification System（MIS）4.5系统进行脂肪酸的比对鉴定。
（2）各微生物群落表征研究认为不同微生物群落的PLFA图谱存在较大差异,而同一类群的微生物PLFA图谱则很相近,因而可以利用某些特征PLFA表征某一类群的微生物^[23]。通常：n14:0-n20:0为普通直链饱和脂肪酸,表征细菌/总微生物量^[23];18:1 ω9c、18:2 ω6c、18:3 ω6c、16:1 ω5c表征真菌（Fungi）群落微生物量^[24,25,26];16:0 10-methyl、17:0 10-methyl、18:0 10-methyl、17:1 w7c 10-methyl表征放线菌（Actinobacteria）群落微生物量^{[24, 27]};14:0 iso、15:0 iso/ anteiso、16:0 iso、17:0 iso/ anteiso、17:1 iso ω9c表征革兰氏阳性细菌（G⁺）群落微生物量^[6];16:1ω7c、16:1ω9c、17:1ω8c、18:1ω7c、17:0 cycle ω7c、19:0 cycle ω7c表征革兰氏阴性细菌（G^-）群落微生物量^[6];饱和直链脂肪酸、革兰氏阳性细菌（G⁺）、革兰氏阴性细菌（G^-）和放线菌微生物量之和表示细菌群落微生物量,同时将所有微生物PLFAs总和表征为总微生物量。
（注：不同PLFA分子常以一系列C原子数目结合字母表示^[28],如18:1 ω6c,18代表脂肪酸含18个C原子,1代表含有一个双键,ω及后面的数字代表双键与脂肪端的距离;脂肪酸的顺势和反式构型分别用字母c和t表示,字母标在后面;有甲基异型存在时用anteiso和iso表示,分别指甲基在脂肪端末尾第3个和第2个C原子上;methyl表示甲基,前面的数字代表甲基距离羧基端距离;cycle表示为环丙烷脂肪酸。）

1.4 数据处理

利用SPSS19.0和 Excel2010软件对数据进行方差分析、显著检验和相关分析并作图。利用Canoco4.5软件对微生物群落PLFA进行冗余分析（redundancy analysis,RDA）并作图。
利用Shannon-Winner多样性指数（H）、Pielou平均度指数（J）和Simpson丰富度指数（D）以PLFA为指征计算微生物多样性。计算公式如下：
H=-ΣP_ilnP_i
J=H/lnS
D = 1-∑P_i²
式中,P_i是第i种磷脂脂肪酸的相对丰度,即i种特征磷脂脂肪酸的含量除以样品中特征磷脂脂肪酸的总含量;S为每个样品中鉴定到的可供分析用的特征PLFA的种类数量。

2 结果

2.1 施用生物炭和炭基肥对土壤理化性质影响

由表2可知不同处理间全钾含量差异不显著（P>0.05）;PMC和BF处理的pH最高,显著高于BIO处理;CS和BF处理的土壤全氮含量最低,显著低于PMC处理,BIO处理居间;BF和BIO处理有机质含量最低,显著低于PMC处理;PMC处理的土壤含水量最高,其他处理间没有显著差异。
Table 2
表2
表2不同处理对土壤理化性质的影响
Table 2Effects of different treatments on soil physic-chemical properties

处理 Treatments	pH	全钾 Total K(g·kg^-1)	全氮 Total N (g·kg^-1)	有机质 Soil Organic matter (g·kg^-1)	土壤含水量 Soil moisture content（%）
CS	5.98±0.19bc	22.72±0.43a	0.90±0.04b	17.18±0.37ab	10.05±0.76b
PMC	6.35±0.12a	22.65±0.11a	1.01±0.05a	18.16±0.74a	11.75±0.49a
BF	6.21±0.32ab	22.52±0.11a	0.90±0.01b	15.86±0.61b	10.00±0.09b
BIO	5.80±0.21c	22.50±0.59a	0.97±0.03ab	15.83±1.02b	9.94±0.67b

Different small letters within one column mean significant difference of Duncan multiple range test among different treatments at 5% level. Data were expressed as means±SE. The same as below每列数据后不同小写字母表示Duncan多重比较不同处理差异显著,数据表示平均值±标准误差。下同

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2.2 施用生物炭和炭基肥对土壤微生物群落结构影响

PMC处理的土壤微生物总PLFAs含量显著高于其他处理（图1）,其余处理间则没有显著差异。细菌PLFAs含量仍以PMC处理最高,BF最低,CS和BIO处理居中且二者间无显著差异。放线菌PLFAs含量以CS处理最低,与PMC处理存在显著差异,BF和BIO处理居间无显著性差异。BIO处理的革兰氏阳性和革兰氏阴性细菌的PLFAs（图2）含量均最低,与PMC处理存在显著差异。

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图1不同处理土壤微生物的特征 PLFAs 含量
-->Fig. 1Content of soil microbial PLFAs under different treatments
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图2不同处理革兰氏阳性与阴性细菌菌落的特征 PLFAs 含量
-->Fig. 2Content of soil G⁺ and G^- microbial PLFAs under different treatments
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不同处理间的Pielou均匀度指数和Simpson优势度指数均没有显著差异（表3）,Shannon-Winner多样性指数BF处理显著高于BIO处理, PMC和CS处理居间没有显著差异。真菌PLFAs/细菌PLFAs比值（图3-a）BIO处理最低,其余3种处理之间没有显著差异,革兰氏阳性细菌PLFAs/革兰氏阴性细菌PLFAs比值（图3-b）BF和PMC处理最低,显著低于BIO处理。
Table 3
表3
表3不同处理土壤微生物多样性指数
Table 3Soil microbial diversity indices under different treatments

处理 Treatment	多样性指数 Shannon-Winner Diversity Index(H)	均匀度指数 Pielou Evenness Index(J)	优势度指数 Simpson Dominance Index(D)
CS	2.89ab	0.88a	0.92a
PMC	2.88ab	0.87a	0.92a
BF	2.91a	0.88a	0.93a
BIO	2.83b	0.86a	0.91a

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图3不同处理土壤中真菌/细菌(a)、革兰氏阳性细菌/革兰氏阴性细菌(b)比
-->Fig. 3Rations of soil fungi /bacteria (a) and G⁺ /G^-(b) under different treatments
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2.3 土壤微生物PLFA与土壤理化指标的关系

以不同PLFA为物种因子对不同理化指标做约束性排序（RDA分析）,利用蒙特卡罗P值检验中有显著贡献（P<0.05）的环境因子与物种因子做物种-环境双序图（图4）,结果显示,第1主轴和第2主轴分别解释了微生物群落结构与理化性质关系总变异的76.3%和6.4%;对PLFA丰度有显著影响的土壤理化指标及重要性排序分别是土壤pH、全氮、有机质、含水量和全钾（表4）,其中含水量和全钾对土壤微生物群落有显著影响（P < 0.05）,pH、全氮和有机质对土壤微生物群落有极显著影响（P < 0.01）。pH和全氮与16:1 ω7c、18:1 ω7c（G^-）、15:0 iso、15:0 anteiso（G⁺）、16:1 ω5c、18:1 ω9c（真菌）和16:0 10-methyl、18:0 10-methyl（放线菌）存在明显正相关,有机质与16:1 ω5c 、18:2 ω6c（真菌）、17:0 cycle ω7c（G^-）、17:0 anteiso（G⁺）相关性较高,含水量和17:1 w7c 10-methyl（放线菌）、16:1 ω9c（真菌）、17:0 iso（G⁺）和17:1 ω8c 、19:0 cycle ω7c（G^-）相关性强,全钾和17:0 10-methyl（放线菌）、16:0 iso、14:0 iso、17:1 iso ω9c（G⁺）和14:0、20:0（细菌）相关性较高。
Table 4
表4
表4理化因子变量解释的重要性排序与显著性检验
Table 4Importance and significance levels of physic-chemical parameters

土壤理化指标 Soil physic-chemical parameters	重要性排序 Importance rank	理化因子所占解释量 Variance explanation of different parameters	P P-value estimate
pH	1	0.703	0.002
全氮TN	2	0.607	0.004
有机质SOM	3	0.509	0.010
土壤含水量MC	4	0.506	0.014
全钾TK	5	0.402	0.048

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图4土壤微生物群落结构和土壤化学性质的RDA分析
-->Fig. 4Redundancy analysis of soil microbial community structure and chemical properties
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相关分析结果表明（表5）,土壤pH与真菌、革兰氏阳性和革兰氏阴性细菌PLFAs含量极显著相关,与放线菌PLFAs显著相关;全氮与革兰氏阳性与阴性细菌PLFAs含量极显著相关,与真菌与放线菌PLFAs含量显著相关;有机质与真菌PLFAs含量极显著相关,与革兰氏阳性与阴性细菌PLFAs含量显著相关;含水量与真菌、放线菌、革兰氏阳性细菌和革兰氏阴性细菌PLFAs含量显著相关;全钾和总PLFAs与细菌PLFAs含量极显著相关,和放线菌与革兰氏阳性细菌PLFAs含量显著相关。
Table 5
表5
表5土壤微生物不同群落与土壤化学性质的相关性
Table 5Correlation between soil microbial composition and physic-chemical properties

	pH	全钾 TK	全氮 TN	有机质 SOM	含水量 MC
总PLFAs Total PLFAs	0.671	0.846^**	0.690	0.639	0.695
细菌PLFAs Bacterial PLFAs	0.573	0.850^**	0.611	0.567	0.636
真菌PLFAs Fungi PLFAs	0.924^**	0.480	0.823^*	0.843^**	0.731^*
放线菌PLFAs Actinomycetes PLFAs	0.834^*	0.795^*	0.823^*	0.672	0.775^*
革兰氏阳性PLFAs G⁺ PLFAs	0.916^**	0.761^*	0.871^**	0.816^*	0.800^*
革兰氏阴性PLFAs G^- PLFAs	0.958^**	0.600	0.860^**	0.785^*	0.764^*

*在0.05 水平上显著相关。**在0.01水平上显著相关 * Mean significant difference at 0.05 level. ** Mean significant difference at 0.01 level

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3 讨论

各有机物料连续施用7年后,土壤理化性质发生了较大变化。与试验前相比（土壤pH5.5,全氮0.86 g·kg^-1,有机质15.3 g·kg^-1,全钾20.40 g·kg^-1）不同处理pH、全氮、有机质和全钾含量均有所提高。生物炭制备过程中矿质元素趋于离子态存在,损失不大,施入土壤后显碱性可以提升土壤酸碱度^[29],其中炭基肥和猪厩肥处理提升幅度最大,这可能是炭基肥和腐熟的猪厩肥施入土壤后不会产生过多中间酸性产物^[4],而生物炭处理施用的生物炭颗粒添加了酸性胶结剂（各有机物料pH见表1）,秸秆也会在土中腐解产生酸性物质^[4]。生物炭对土壤全氮含量的提高是由于在烧制过程中氮的状态发生了转变^[30],出现较多以C-N杂环稳定形态存在的氮^[31],施入土壤能够长时间保持稳定不易被分解^[32],与此相似,在碳素每年投入不等量的情况下（CS、BIO和BF分别为1 999.8、499.8和57.98 kg·hm^-2,有机物料含碳量见表1）,生物炭和炭基肥处理有效提高土壤有机质含量且在长期施用中与CS处理达到相同水平。
植物种类、土壤性质以及土壤的管理方式是影响土壤微生物的主要因素^[33]。本试验中PMC处理的各种微生物群落生物量均较为丰富,主要的原因有：腐熟的猪厩肥含有大量活性微生物,对于土壤起到“接种”的作用^[34];猪厩肥本身养分（主要是碳、氮）相较秸秆和生物炭更适宜微生物的繁殖^[7]。生物炭对微生物的影响主要有两方面,一是生物炭本身多孔性的微观结构和巨大比表面积可为微生物提供适宜生境^[15],二是生物炭除了本身能提供少量营养物质外还可以通过吸附养分或改良土壤性质间接影响微生物群落^[13]。细菌一般是土壤微生物中的优势群落^[6],本试验中不同处理细菌PLFAs占总PLFAs含量的比值均在75%以上,生物炭对微生物的影响首先体现在细菌群落中,炭基肥处理由于所含生物炭量不高（57.98 kg·hm^-2）,对土壤细菌微生物群落的影响不及生物炭处理（499.8 kg·hm^-2）显著。细菌和真菌对土壤有机物质的分解途径不同,真菌易生存于低营养、难分解以及低氮含量有机物的环境,底物循环时间偏长;细菌则相反,偏好生存在营养丰富、有机物易于分解的土壤中,底物周转快^[35]。本研究中生物炭处理的真菌PLFAs含量最低,相较猪厩肥处理不利于真菌积累。可能的原因是真菌生长并不过多依赖生物炭孔隙和生物炭带来的无机营养,而试验所用的玉米芯生物炭也没能提供足够数量的适宜真菌生存的孔隙^[15]。JIN等^[36]施用生物炭的改土试验还曾发现过真菌遗传多样性和数量减少的现象。
微生物群落结构关乎生态系统的稳定和健康^{[6, 37]},FTDE等^[38]的研究发现,真菌/细菌生物量比值与农田生态系统稳定性相关,一些研究^[42]也表明真菌/细菌生物量比值与微生物多样性存在相关性。本研究中炭基肥处理的Shannon-Winner多样性指数和真菌/细菌比值均最高,显著高于生物炭处理,说明相较生物炭处理施用炭基肥更有利于提高土壤微生物的群落结构多样性,生物炭处理相较炭基肥处理更有利于细菌群落PLFAs生物量的生长是引起这一差异的原因。支链饱和脂肪酸与环化单烯等不饱和脂肪酸通常分别是革兰氏阳性和阴性细菌的特征标记物^[25],其比值也常用来表示土壤微生物群落结构^[39]及与环境的关系^{[38, 40]},比值高低与环境胁迫程度有关^[20]。本研究中生物炭处理的G⁺ /G^-比值最高,说明施用生物炭对土壤微生物的胁迫程度较施用猪厩肥和炭基肥要高一些。但生物炭处理的革兰氏阳性和革兰氏阴性细菌PLFAs含量均低于猪厩肥处理,说明相比猪厩肥,生物炭处理更不利于革兰氏阴性细菌的积累,这与STEINBEISS等^[12]的研究施入生物炭能增加土壤革兰氏阴性细菌生物量的结论似有不符,但本试验中炭基肥处理却没有这种现象,可能的原因是炭基肥和生物炭处理施入土壤的纯生物质炭量不同（BIO和BF分别为499.8 kg·hm^-2和57.98 kg·hm^-2）,导致不同处理土壤微生物受胁迫程度不同。
土壤理化性质如pH、含水量、有机质和土壤养分也是影响土壤微生物群落的重要因素^{[20, 41]}。本研究RDA分析结果显示,pH、全氮和有机质对土壤微生物影响最大,呈极显著相关,这与许多研究结果^{[6-7, 42]}类似,pH对微生物适宜生存环境影响巨大^[2],氮素是微生物必不可少的养分元素^[1],而有机质是土壤养分最重要的衡量指标,说明这三种因素是影响本试验微生物群落的关键因子,其他显著影响因子还包括土壤含水量与全钾。但在重要性排序上pH位于全氮和有机质之前,此结果似乎和一些文献^[7]相冲突,这主要有以下几方面原因：首先全氮和有机质并不是对所有微生物群落有显著影响,由表5可知全氮和有机质只是与部分微生物群落极显著相关,全氮与革兰氏阳性与阴性细菌,有机质与真菌,而土壤pH与真菌、革兰氏阳性和革兰氏阴性细菌均极显著相关;其次生物炭等有机物料拥有巨大的比表面积,pH的变化会对生物炭表面可变负电荷产生影响,造成铵态氮和磷素等营养元素的吸附障碍从而影响其对土壤微生物的有效性^[2]。
生物炭被认为是改善土壤肥力和土壤生态,可通过固碳减缓气候变化的有效手段^[10,18],其对土壤的影响机理^[4,8,10,21]多被解释为通过提高土壤pH和吸附作用改善土壤养分利用情况,同时改变土壤微生物群落组成及丰度^[12,18-19],进而对土壤养分循环或理化性质产生作用^[11,12],最终影响作物生长^[11]。这些研究多数集中在土壤性质或微生物分化差异上,关于施用生物炭的土壤在功能微生物作用下养分变异过程的内涵并没有系统深入阐述。现有的研究^{[3-4,8-12,18]}为未来关于生物炭对土壤微生物群落功能性的研究提供了广阔思路,如不同生物炭与土壤不同养分及相关酶与功能微生物的互作,生物炭与更广泛的微生物（土壤动物等）功能性研究,除本文研究的炭基肥外的生物炭菌剂等生物炭功能性产品,生物炭微生物环境风险等方面^[18]都值得我们去探索。

4 结论

长期施用生物炭和炭基肥可以改善土壤的理化性质。相较猪厩肥,施用生物炭不利于真菌和革兰氏阴性细菌群落生物量的积累,且施用生物炭和炭基肥对土壤微生物群落的影响不同,施用生物炭可以显著提高土壤细菌生物量,施用炭基肥能显著提高土壤微生物Shannon-Winner多样性指数和真菌/细菌比以及降低革兰氏阳性细菌/革兰氏阴性细菌比,有利于土壤微生物群落结构多样性提高。
长期应用生物炭等有机物料培肥改土后,土壤理化指标对微生物群落产生显著影响的因子及重要性依次为pH、全氮、有机质、含水量和全钾,其中前3种因素的影响极显著。
The authors have declared that no competing interests exist.

参考文献原文顺序
文献年度倒序
文中引用次数倒序
被引期刊影响因子

[1]	徐阳春, 沈其荣, 冉炜. 长期免耕与施用有机肥对土壤微生物生物量碳、氮、磷的影响 . 土壤学报, 2002, 39(01): 83-90.https://doi.org/10.11766/trxb200103110113 URL [本文引用: 3]摘要 The effects of tillage and manure application on soil microbial biomass carbon(MBC),microbial biomass nitrogen(MBN),and microbial biomass phosphorus(MBP)were investigated in a field experiment,which consisted of two main treatments of conventional tillage and zero-tillage and five sub-treatments of control(no fertilizers),chemical fertilizers,chemical fertilizers plus straw,chemical fertilizers plus green manure,and chemical fertilizers plus pig manure and had continued for 16 years at the farm of Jurong Agricultural Science Institute.The crop rotation of the experiment was summer rice(Oryza Sativa L.)and winter wheat(Triticum aestivum).The results showed that MBC and MBN in zero-tillage treatment increased by 25.4% and 45.4%,respectively compared to those in conventional tillage treatment in 0~5cm depth layer after 16 years.However,there were no significant differences of MBC,MBN,and MBP in subsoil(5~10cm)between conventional tillage and zero-tillage.Although the application rate of N,P,and K in each sub-treatment were equal,the contents of MBC,MBN,and MBP in 0~5cm and 5~10cm depth layer followed the order in the treatments:fertilizers plus pig manure>fertilizers plus straw>fertilizers plus green manure>chemical fertilizers>control. XU Y, C,SHEN Q R, RAN W.Effects of zero-tillage and application of manure on soil microbial biomass C,N and P after sixteen years of cropping .Acta Pedologica Sinica, 2002, 39(01): 83-90. (in Chinese)https://doi.org/10.11766/trxb200103110113 URL [本文引用: 3]摘要 The effects of tillage and manure application on soil microbial biomass carbon(MBC),microbial biomass nitrogen(MBN),and microbial biomass phosphorus(MBP)were investigated in a field experiment,which consisted of two main treatments of conventional tillage and zero-tillage and five sub-treatments of control(no fertilizers),chemical fertilizers,chemical fertilizers plus straw,chemical fertilizers plus green manure,and chemical fertilizers plus pig manure and had continued for 16 years at the farm of Jurong Agricultural Science Institute.The crop rotation of the experiment was summer rice(Oryza Sativa L.)and winter wheat(Triticum aestivum).The results showed that MBC and MBN in zero-tillage treatment increased by 25.4% and 45.4%,respectively compared to those in conventional tillage treatment in 0~5cm depth layer after 16 years.However,there were no significant differences of MBC,MBN,and MBP in subsoil(5~10cm)between conventional tillage and zero-tillage.Although the application rate of N,P,and K in each sub-treatment were equal,the contents of MBC,MBN,and MBP in 0~5cm and 5~10cm depth layer followed the order in the treatments:fertilizers plus pig manure>fertilizers plus straw>fertilizers plus green manure>chemical fertilizers>control.
[2]	李春越, 王益, BROOKES P, 党廷辉, 王万忠. pH对土壤微生物C/P比的影响 . 中国农业科学, 2013, 46(13): 2709-2716.https://doi.org/10.3864/j.issn.0578-1752.2013.13.009 Magsci [本文引用: 3]摘要【目的】了解pH对土壤微生物量的影响，揭示磷素转化机理及提高磷素利用率的生物调控措施。【方法】以英国洛桑研究所长期定位试验pH梯度土壤为研究对象，研究不同pH下，耕地土壤微生物碳磷比（C/P）及磷素有效性的差异。【结果】微生物C/P比与土壤pH、全碳、无机磷含量以及磷素回收率之间具有一定的相关性。磷素回收率和微生物C/P比随pH的增加而增加，土壤全碳、全磷、C/N和微生物量磷则随pH的增加而减少。土壤呼吸强度随培养时间延长呈线性增加。微生物量碳和ATP呈现出明显的正相关，相关系数高达0.912（n=16）。【结论】pH对于磷素的利用起着重要的作用。微生物C/P比可以作为土壤磷素利用的一个重要指示指标，微生物量C/P比的大小反映土壤微生物对土壤磷有效性的调节作用。 LI C Y, WANG Y, BROOKES P, DANG T H, WANG W Z.Effect of soil pH on soil microbial carbon phosphorus ratio .Scientia Agricultura Sinica, 2013, 46(13): 2709-2716. (in Chinese)https://doi.org/10.3864/j.issn.0578-1752.2013.13.009 Magsci [本文引用: 3]摘要【目的】了解pH对土壤微生物量的影响，揭示磷素转化机理及提高磷素利用率的生物调控措施。【方法】以英国洛桑研究所长期定位试验pH梯度土壤为研究对象，研究不同pH下，耕地土壤微生物碳磷比（C/P）及磷素有效性的差异。【结果】微生物C/P比与土壤pH、全碳、无机磷含量以及磷素回收率之间具有一定的相关性。磷素回收率和微生物C/P比随pH的增加而增加，土壤全碳、全磷、C/N和微生物量磷则随pH的增加而减少。土壤呼吸强度随培养时间延长呈线性增加。微生物量碳和ATP呈现出明显的正相关，相关系数高达0.912（n=16）。【结论】pH对于磷素的利用起着重要的作用。微生物C/P比可以作为土壤磷素利用的一个重要指示指标，微生物量C/P比的大小反映土壤微生物对土壤磷有效性的调节作用。
[3]	陈温福, 张伟明, 孟军. 农用生物炭研究进展与前景 . 中国农业科学, 2013, 46(16): 3324-3333.https://doi.org/10.3864/j.issn.0578-1752.2013.16.003 URL Magsci [本文引用: 6]摘要生物炭以其良好的解剖结构和理化性质，广泛的材料来源和广阔的产业化发展前景，成为当今农业、能源与环境等领域的研究热点。本文综合分析、评述了生物炭在土壤、作物、农田生态系统等领域应用的主要研究进展及其未来保障中国粮食安全的重要意义，从低碳、循环、可持续视角，客观、辩证地探讨了生物炭在农业上的应用价值及其产业化发展前景。生物炭在修复土壤障碍，提升耕地生产性能和作物生产能力，促进农业可持续发展和保障国家粮食安全等方面具有重要现实意义和应用价值，本文结合中国国情，提出了进一步深入研究与开发生物炭产业的方向与建议，旨在为中国生物炭产业的健康发展提供参考。 CHEN W F, ZHANG W M, MENG J.Advances and prospects in research of biochar utilization in agriculture .Scientia Agricultura Sinica, 2013, 46(16): 3324-3333. (in Chinese)https://doi.org/10.3864/j.issn.0578-1752.2013.16.003 URL Magsci [本文引用: 6]摘要生物炭以其良好的解剖结构和理化性质，广泛的材料来源和广阔的产业化发展前景，成为当今农业、能源与环境等领域的研究热点。本文综合分析、评述了生物炭在土壤、作物、农田生态系统等领域应用的主要研究进展及其未来保障中国粮食安全的重要意义，从低碳、循环、可持续视角，客观、辩证地探讨了生物炭在农业上的应用价值及其产业化发展前景。生物炭在修复土壤障碍，提升耕地生产性能和作物生产能力，促进农业可持续发展和保障国家粮食安全等方面具有重要现实意义和应用价值，本文结合中国国情，提出了进一步深入研究与开发生物炭产业的方向与建议，旨在为中国生物炭产业的健康发展提供参考。
[4]	战秀梅, 彭靖, 王月, 刘轶飞, 陈坤, 韩晓日, 王恒飞, 蔺文成, 李喜研. 生物炭及炭基肥改良棕壤理化性状及提高花生产量的作用 . 植物营养与肥料学报, 2015, 21(6): 1633-1641.https://doi.org/10.11674/zwyf.2015.0631 URL Magsci [本文引用: 7]摘要【目的】炭基复合肥是生物炭农用的另一种方式,生物炭作为土壤改良剂对土壤改良研究的报道较多,但大多为短期培养或模拟试验。目前更缺乏生物炭与传统土壤培肥方式的比较研究。本研究旨在通过4年的田间微区定位试验,开展生物炭及炭基复合肥对棕壤理化性质及花生产量的影响研究,以期为生物炭的培肥改土及合理农用提供理论依据。【方法】定位试验于2009年开始连续4年进行了花生微区田间试验（2 m~2）。试验设4个处理分别为秸秆还田＋NPK（CS）、施用猪厩肥＋NPK（PMC）、生物炭＋NPK（BIO）和基于生物炭的炭基复合肥（BF）所有处理均为等氮磷钾养分,BIO处理与PMC处理为等碳量,BIO处理相当于CS处理所施用的玉米秸秆量制备得到的生物炭量,BF处理碳含量低于BIO碳含量每个处理重复3次,随机排列。分析试验前和2012年收获后土壤理化性质,比较各处理4年的花生产量。【结果】连续施用4年后,与试验前相比,BIO处理的土壤有机碳提高了27.6%全氮含量提高了75.6%,显著高于其他各处理,土壤pH提高了0.14个单位,显著高于CS处理,与PMC处理相近;土壤碱解氮、速效磷、速效钾和CEC值与CS或PMC处理相近;BIO处理的土壤毛管孔隙度和田间持水量显著高于其他处理容重和土壤总孔隙度与CS和PMC处理差异不显著;4年中花生产量均居首位,从3198.5 kg/hm~2提高到4818.0 kg/hm~2,但与PMC处理差异不显著。连续施用4年后,BF处理土壤pH较试验前提高了0.57个单位显著高于其他各处理,优势显著;土壤有机碳和全氮含量较试验前分别提高了4.4%和27.9%显著低于BIO处理对土壤物理性质的调节作用也不及BIO处理其他指标差异不显著但总体上与CS或PMC处理相近;BF处理的花生产量在试验的前3年与BIO处理差异不显著,第4年较BIO处理降低了317.1 kg/hm2,差异显著介于PMC和CS处理之间。【结论】各处理作物17 ZHAN X M, PENG J, WANG Y, LIU Y F, CHEN K, HAN X R, WANG H F, LIN W C, LIX Y.Influences of application of biochar and biochar-based fertilizr on brown soil physiochemical properties and peanut yields .Journal of Plant Nutrition and Fertilizer, 2015, 21(6): 1633-1641. (in Chinese)https://doi.org/10.11674/zwyf.2015.0631 URL Magsci [本文引用: 7]摘要【目的】炭基复合肥是生物炭农用的另一种方式,生物炭作为土壤改良剂对土壤改良研究的报道较多,但大多为短期培养或模拟试验。目前更缺乏生物炭与传统土壤培肥方式的比较研究。本研究旨在通过4年的田间微区定位试验,开展生物炭及炭基复合肥对棕壤理化性质及花生产量的影响研究,以期为生物炭的培肥改土及合理农用提供理论依据。【方法】定位试验于2009年开始连续4年进行了花生微区田间试验（2 m~2）。试验设4个处理分别为秸秆还田＋NPK（CS）、施用猪厩肥＋NPK（PMC）、生物炭＋NPK（BIO）和基于生物炭的炭基复合肥（BF）所有处理均为等氮磷钾养分,BIO处理与PMC处理为等碳量,BIO处理相当于CS处理所施用的玉米秸秆量制备得到的生物炭量,BF处理碳含量低于BIO碳含量每个处理重复3次,随机排列。分析试验前和2012年收获后土壤理化性质,比较各处理4年的花生产量。【结果】连续施用4年后,与试验前相比,BIO处理的土壤有机碳提高了27.6%全氮含量提高了75.6%,显著高于其他各处理,土壤pH提高了0.14个单位,显著高于CS处理,与PMC处理相近;土壤碱解氮、速效磷、速效钾和CEC值与CS或PMC处理相近;BIO处理的土壤毛管孔隙度和田间持水量显著高于其他处理容重和土壤总孔隙度与CS和PMC处理差异不显著;4年中花生产量均居首位,从3198.5 kg/hm~2提高到4818.0 kg/hm~2,但与PMC处理差异不显著。连续施用4年后,BF处理土壤pH较试验前提高了0.57个单位显著高于其他各处理,优势显著;土壤有机碳和全氮含量较试验前分别提高了4.4%和27.9%显著低于BIO处理对土壤物理性质的调节作用也不及BIO处理其他指标差异不显著但总体上与CS或PMC处理相近;BF处理的花生产量在试验的前3年与BIO处理差异不显著,第4年较BIO处理降低了317.1 kg/hm2,差异显著介于PMC和CS处理之间。【结论】各处理作物17
[5]	陈温福, 韩晓日, 徐正进, 刘金. 一种炭基缓释花生专用肥料及其制备方法:CN101121619. 2008-02-13. [本文引用: 1] CHEN W F, HAN X R, XU Z J, LIU J. Biochar-based slow release peanut fertilizer and preparation method:CN101121619 .2008-02-13. (in Chinese) [本文引用: 1]
[6]	王巍巍, 赵琼, 赵欣然, 曾德慧, 艾桂艳. 凋落物管理对樟子松人工林土壤微生物群落结构的影响 . 生态学杂志, 2015, 34(9): 2605-2612.URL Magsci [本文引用: 6]摘要采用磷脂脂肪酸法(PLFA)分析短期(18个月)凋落物管理对樟子松人工林土壤微生物群落结构的影响.结果表明,凋落物移除对土壤各类微生物的PLFAs均无显著影响,但凋落物加倍显著改变了土壤微生物群落结构.凋落物加倍显著提高了土壤PLFAs总量、革兰氏阳性细菌(G+)与革兰氏阴性细菌(G一)和原生动物PLFAs,但对真菌和放线菌PLFAs以及细菌/真菌值、G+/G-值影响不显著.凋落物加倍使土壤总PLFAs上升了82.4％,细菌PLFAs上升了93.9％.凋落物管理对樟子松人工林土壤微生物的Shannon多样性指数、Pielou均匀度指数和Simpson优势度指数均无显著影响.主成分分析表明,主成分1 (89.83％)上得分系数较高的PLFAs多为细菌的特征PLFAs,主要有16:1ω5c、16:1ω7c、cy17:0ω7c、cy19:0ω7c、i14:0、i15:0. WANG W W, ZHAO Q, ZHAO X R, ZENG D H, AI G Y.Effects of litter manipulation on soil microbial community structure in a Pinus sylvestris var.mongolica plantation .Chinese Journal of Ecology, 2015, 34(9): 2605-2612. (in Chinese)URL Magsci [本文引用: 6]摘要采用磷脂脂肪酸法(PLFA)分析短期(18个月)凋落物管理对樟子松人工林土壤微生物群落结构的影响.结果表明,凋落物移除对土壤各类微生物的PLFAs均无显著影响,但凋落物加倍显著改变了土壤微生物群落结构.凋落物加倍显著提高了土壤PLFAs总量、革兰氏阳性细菌(G+)与革兰氏阴性细菌(G一)和原生动物PLFAs,但对真菌和放线菌PLFAs以及细菌/真菌值、G+/G-值影响不显著.凋落物加倍使土壤总PLFAs上升了82.4％,细菌PLFAs上升了93.9％.凋落物管理对樟子松人工林土壤微生物的Shannon多样性指数、Pielou均匀度指数和Simpson优势度指数均无显著影响.主成分分析表明,主成分1 (89.83％)上得分系数较高的PLFAs多为细菌的特征PLFAs,主要有16:1ω5c、16:1ω7c、cy17:0ω7c、cy19:0ω7c、i14:0、i15:0.
[7]	郭芸, 孙本华, 王颖, 魏静, 高明霞, 张树兰, 杨学云. 长期施用不同肥料土plfa指纹特征 . 中国农业科学, 2017, 50(1): 94-103.https://doi.org/10.3864/j.issn.0578-1752.2017.01.009 URL [本文引用: 4]摘要【目的】土壤微生物群落结构的组成与活性的变化是衡量土壤肥力的重要指标,研究长期不同施肥和土壤管理方式对塿土微生物群落结构的影响,对于指导塿土施肥和土壤管理,实现农田可持续利用具有重要意义。【方法】以陕西杨凌"国家黄土肥力与肥料效益监测基地"长期肥料定位试验为基础,运用磷脂脂肪酸标记法(PLFA),研究了塿土长期不同施肥及土地利用方式下土壤微生物群落结构及其与土壤理化性质的关系。处理包括:长期不施肥(CK)、单施氮肥(N)、长期配合施用氮钾(NK)、磷钾(PK)、氮磷(NP)、有机肥和氮磷钾(MNPK)以及长期休闲(FL)和撂荒(AB)。【结果】与对照相比,MNPK、NP和撂荒处理土壤总PLFA分别增加218.8%、73.9%和74.3%,细菌分别增加188.3%、80.8%和82.6%,真菌分别增加了315.8%、111.5%和167.0%,放线菌分别增加了23.7%、21.3%和16.3%,同时也显著增加了真菌/细菌比;N、NK和PK土壤总PLFA、细菌、真菌差异不显著,但PK显著降低放线菌的含量;与农田施肥相比,休闲和撂荒显著降低G~+和G~-含量。多样性指数结果表明,长期有机无机配施明显提高土壤微生物群落的Shannon-Winner多样性指数、Simpson优势度和Pielou均匀度指数,撂荒和NP也能显著增加Shannon-Winner多样性指数和Pielou均匀度指数,而长期休闲处理均明显降低了这些指数。主成分分析表明,MNPK、NP、撂荒和休闲土壤微生物群落结构发生较大变化;MNPK显著提高G~-(18:1ω5c,cy19:0ω7c)、细菌(16:0、10Me22:0饱和脂肪酸)及真核生物(18:3ω6c、16:3ω6c,22:2ω6c)的多度值,撂荒(AB)和NP显著提高细菌(15:0,18:0,22:0,17:0饱和脂肪酸)的多度值。RDA分析表明,土壤理化性质对微生物菌群影响的重要性依次为有机质全氮含水量速效磷pH容重速效钾,这些理化因子均是微生物生长的关键因子。【结论】长期有机无机肥配施、氮磷配施和撂荒提高了土壤微生物群落结构多样性,从而改善了土壤生态环境,而长期休闲不利于土壤生态系统的稳定和健康。 GUO Y, SUN B H, WANG Y, WEI J, GAO M X, ZHANG S L, YANG X Y.PLFA fingerprint characteristics of an anthropogenic loess soil under long-term different fertilizations .Scientia Agricultura Sinica, 2017, 50(1): 94-103. (in Chinese)https://doi.org/10.3864/j.issn.0578-1752.2017.01.009 URL [本文引用: 4]摘要【目的】土壤微生物群落结构的组成与活性的变化是衡量土壤肥力的重要指标,研究长期不同施肥和土壤管理方式对塿土微生物群落结构的影响,对于指导塿土施肥和土壤管理,实现农田可持续利用具有重要意义。【方法】以陕西杨凌"国家黄土肥力与肥料效益监测基地"长期肥料定位试验为基础,运用磷脂脂肪酸标记法(PLFA),研究了塿土长期不同施肥及土地利用方式下土壤微生物群落结构及其与土壤理化性质的关系。处理包括:长期不施肥(CK)、单施氮肥(N)、长期配合施用氮钾(NK)、磷钾(PK)、氮磷(NP)、有机肥和氮磷钾(MNPK)以及长期休闲(FL)和撂荒(AB)。【结果】与对照相比,MNPK、NP和撂荒处理土壤总PLFA分别增加218.8%、73.9%和74.3%,细菌分别增加188.3%、80.8%和82.6%,真菌分别增加了315.8%、111.5%和167.0%,放线菌分别增加了23.7%、21.3%和16.3%,同时也显著增加了真菌/细菌比;N、NK和PK土壤总PLFA、细菌、真菌差异不显著,但PK显著降低放线菌的含量;与农田施肥相比,休闲和撂荒显著降低G~+和G~-含量。多样性指数结果表明,长期有机无机配施明显提高土壤微生物群落的Shannon-Winner多样性指数、Simpson优势度和Pielou均匀度指数,撂荒和NP也能显著增加Shannon-Winner多样性指数和Pielou均匀度指数,而长期休闲处理均明显降低了这些指数。主成分分析表明,MNPK、NP、撂荒和休闲土壤微生物群落结构发生较大变化;MNPK显著提高G~-(18:1ω5c,cy19:0ω7c)、细菌(16:0、10Me22:0饱和脂肪酸)及真核生物(18:3ω6c、16:3ω6c,22:2ω6c)的多度值,撂荒(AB)和NP显著提高细菌(15:0,18:0,22:0,17:0饱和脂肪酸)的多度值。RDA分析表明,土壤理化性质对微生物菌群影响的重要性依次为有机质全氮含水量速效磷pH容重速效钾,这些理化因子均是微生物生长的关键因子。【结论】长期有机无机肥配施、氮磷配施和撂荒提高了土壤微生物群落结构多样性,从而改善了土壤生态环境,而长期休闲不利于土壤生态系统的稳定和健康。
[8]	何绪生, 耿增超, 佘雕, 张保健, 高海英. 生物炭生产与农用的意义及国内外动态 . 农业工程学报, 2011, 27(2): 1-7.URL Magsci [本文引用: 3]摘要近年来，生物炭作为土壤改良剂、肥料缓释载体及碳封存剂备受重视。生物炭在土壤中能够保持数百年至数千年，实现碳的封存固定，生物炭还可以改善土壤理化性质及微生物的活性，培肥土壤肥力，延缓肥料养分释放，降低肥料及土壤养分的损失，减轻土壤污染。生物质的热裂解及气化均可产生生物炭，但是慢速热裂解和热水炭化工艺的生物炭产率最大，同时还可获得生物油及混合气，生物油及混合气可升级加工为氢气、生物柴油或化学品，这有助于减轻对化石能源或原料的依赖。生物炭的生产及农用是碳减排的过程，废弃生物质生产生物炭及其农用的效益是多赢的。国外在废弃生物质热裂解生产生物炭及农用方面做了许多研究工作。中国在生物质热裂解获得生物能源方面做了较多工作，但对生物炭的生产及农用重视不够。今后，中国应以废弃生物质生产生物炭，并将生物炭农用作为生物能源、环境及农业可持续发展的战略。 HE X S, GENG Z C, SHE D, ZHANG B J, GAO H Y.Implications of production and agricultural utilization of biochar andits international dynamic .Transactions of the CSAE, 2011, 27(2): 1-7. (in Chinese)URL Magsci [本文引用: 3]摘要近年来，生物炭作为土壤改良剂、肥料缓释载体及碳封存剂备受重视。生物炭在土壤中能够保持数百年至数千年，实现碳的封存固定，生物炭还可以改善土壤理化性质及微生物的活性，培肥土壤肥力，延缓肥料养分释放，降低肥料及土壤养分的损失，减轻土壤污染。生物质的热裂解及气化均可产生生物炭，但是慢速热裂解和热水炭化工艺的生物炭产率最大，同时还可获得生物油及混合气，生物油及混合气可升级加工为氢气、生物柴油或化学品，这有助于减轻对化石能源或原料的依赖。生物炭的生产及农用是碳减排的过程，废弃生物质生产生物炭及其农用的效益是多赢的。国外在废弃生物质热裂解生产生物炭及农用方面做了许多研究工作。中国在生物质热裂解获得生物能源方面做了较多工作，但对生物炭的生产及农用重视不够。今后，中国应以废弃生物质生产生物炭，并将生物炭农用作为生物能源、环境及农业可持续发展的战略。
[9]	WOOLF D, AMONETTE J E, STREETPERROT F A, LEHMANN J, JOSEPH S.Sustainable biochar to mitigate global climate change .Nature Communications, 2010, 1(5): 56. DOI:https://www.chinaagrisci.com/article/2018/0578-1752/10.1038/ncomms1053.URLPMID:2964457 [本文引用: 1]摘要 Abstract Production of biochar (the carbon (C)-rich solid formed by pyrolysis of biomass) and its storage in soils have been suggested as a means of abating climate change by sequestering carbon, while simultaneously providing energy and increasing crop yields. Substantial uncertainties exist, however, regarding the impact, capacity and sustainability of biochar at the global level. In this paper we estimate the maximum sustainable technical potential of biochar to mitigate climate change. Annual net emissions of carbon dioxide (CO(2)), methane and nitrous oxide could be reduced by a maximum of 1.8090009Pg CO(2)-C equivalent (CO(2)-C(e)) per year (12% of current anthropogenic CO(2)-C(e) emissions; 1090009Pg=1090009Gt), and total net emissions over the course of a century by 130090009Pg CO(2)-C(e), without endangering food security, habitat or soil conservation. Biochar has a larger climate-change mitigation potential than combustion of the same sustainably procured biomass for bioenergy, except when fertile soils are amended while coal is the fuel being offset.
[10]	LEHMANN J, GAUNT J, RONDON M, READ P.Bio-char sequestration in terrestrial ecosystems - a review .Mitigation and Adaptation Strategies for Global Change, 2006, 11(2): 395-419.https://doi.org/10.1007/s11027-005-9006-5 URL Magsci [本文引用: 3]摘要 The application of bio-char (charcoal or biomass-derived black carbon (C)) to soil is proposed as a novel approach to establish a significant, long-term, sink for atmospheric carbon dioxide in terrestrial ecosystems. Apart from positive effects in both reducing emissions and increasing the sequestration of greenhouse gases, the production of bio-char and its application to soil will deliver immediate benefits through improved soil fertility and increased crop production. Conversion of biomass C to bio-char C leads to sequestration of about 50% of the initial C compared to the low amounts retained after burning (3%) and biological decomposition (< 10–20% after 5–10 years), therefore yielding more stable soil C than burning or direct land application of biomass. This efficiency of C conversion of biomass to bio-char is highly dependent on the type of feedstock, but is not significantly affected by the pyrolysis temperature (within 350–500 66 C common for pyrolysis). Existing slash-and-burn systems cause significant degradation of soil and release of greenhouse gases and opportunies may exist to enhance this system by conversion to slash-and-char systems. Our global analysis revealed that up to 12% of the total anthropogenic C emissions by land use change (0.21 Pg C) can be off-set annually in soil, if slash-and-burn is replaced by slash-and-char. Agricultural and forestry wastes such as forest residues, mill residues, field crop residues, or urban wastes add a conservatively estimated 0.16 Pg C yr 611 . Biofuel production using modern biomass can produce a bio-char by-product through pyrolysis which results in 30.6 kg C sequestration for each GJ of energy produced. Using published projections of the use of renewable fuels in the year 2100, bio-char sequestration could amount to 5.5–9.5 Pg C yr 611 if this demand for energy was met through pyrolysis, which would exceed current emissions from fossil fuels (5.4 Pg C yr 611 ). Bio-char soil management systems can deliver tradable C emissions reduction, and C sequestered is easily accountable, and verifiable.
[11]	ASAI H, SAMSON B K, STEPHAN H M, SONGYIKHANGSUTHOR K, HOMMA K, KIYONO Y, INOUE Y, SHIRAIWA T, HOEIR T.Biochar amendment techniques for upland rice production in Northern Laos: 1. Soil physical properties, leaf SPAD and grain yield .Field Crops Research, 2009, 111(1/2): 81-84.https://doi.org/10.1016/j.fcr.2008.10.008 URL [本文引用: 3]摘要 The objective of this study was to investigate the effect of biochar application (CA) on soil physical properties and grain yields of upland rice ( Oryza sativa L.) in northern Laos. During the 2007 wet season, three different experiments were conducted under upland conditions at 10 sites, combining variations in CA amounts (0–16 t ha 611), fertilizer application rates (N and P) and rice cultivars (improved and traditional) in northern Laos. CA improved the saturated hydraulic conductivity of the top soil and the xylem sap flow of the rice plant. CA resulted in higher grain yields at sites with low P availability and improved the response to N and NP chemical fertilizer treatments. However, CA reduced leaf SPAD values, possibly through a reduction of the availability of soil nitrogen, indicating that CA without additional N fertilizer application could reduce grain yields in soils with a low indigenous N supply. These results suggest that CA has the potential to improve soil productivity of upland rice production in Laos, but that the effect of CA application is highly dependent on soil fertility and fertilizer management.
[12]	STEINBEISS S, GLEIXNER G, ANTONIETTI M.Effect of biochar amendment on soil carbon balance and soil microbial activity .Soil Biology and Biochemistry, 2009, 41(6): 1301-1310.https://doi.org/10.1016/j.soilbio.2009.03.016 URL [本文引用: 5]摘要 We investigated the behavior of biochars in arable and forest soil in a greenhouse experiment in order to prove that these amendments can increase carbon storage in soils. Two qualities of biochar were produced by hydrothermal pyrolysis from 13C labeled glucose (0% N) and yeast (5% N), respectively. We quantified respiratory losses of soil and biochar carbon and calculated mean residence times of the biochars using the isotopic label. Extraction of phospholipid fatty acids from soil at the beginning and after 4 months of incubation was used to quantify changes in microbial biomass and to identify microbial groups utilizing the biochars. Mean residence times varied between 4 and 29 years, depending on soil type and quality of biochar. Yeast-derived biochar promoted fungi in the soil, while glucose-derived biochar was utilized by Gram-negative bacteria. Our results suggest that residence times of biochar in soils can be manipulated with the aim to “design” the best possible biochar for a given soil type.
[13]	PIETIK?INEN J, KIIKKIl? O, FRITZE H. Charcoal as a habitat for microbes and its effect on the microbial community of the underlying humus .Oikos, 2000, 89(2): 231-242.https://doi.org/10.1034/j.1600-0706.2000.890203.x URL [本文引用: 2]摘要 Wildfires produce a charcoal layer, which has an adsorbing capacity resembling activated carbon. After the fire a new litter layer starts to accumulate on top of the charcoal layer, which liberates water-soluble compounds that percolate through the charcoal and the unburned humus layer. We first hypothesized that since charcoal has the capacity to adsorb organic compounds it may form a new habitat for microbes, which decompose the adsorbed compounds. Secondly, we hypothesized that the charcoal may cause depletion of decomposable organic carbon in the underlying humus and thus reduce the microbial biomass. To test our hypotheses we prepared microcosms, where we placed non-heated humus and on top one of the adsorbents: non-adsorptive pumice (Pum), charcoal from Empetrum nigrum (EmpCh), charcoal from humus (HuCh) or activated carbon (ActC). We watered them with birch leaf litter extract. The adsorbing capacity increased in the order Pum<HuCh<EmpCh<ActC, the adsorbents being capable of removing 0%, 26%, 42% and 51% of the dissolved C org in the litter extract, respectively. After one month, all adsorbents harboured microbes, but their amount and basal respiration was largest in EmpCh and HuCh, and smallest in Pum. In addition, different kinds of microbial communities with respect to their phospholipid fatty acid and substrate utilization patterns were formed in the adsorbents. The amount of microbial biomass and number of bacteria did not differ between humus under different adsorbents, although different microbial communities developed in humus under EmpCh compared with Pum, which is obviously related to the increased pH of the humus under EmpCh, and also ActC. We suggest that charcoal from burning can support microbial communities, which are small in size but have a higher specific growth rate than those of the humus. Although the charcoal layer induces changes in the microbial community of the humus, it does not reduce the amount of humus microbes.
[14]	RIVERA-UTRILLA J, BAUTISTA-TOLEDO I,FERRO-GARCIA M A, MORENO-CASTILLA C. Activated carbon surface modifications by adsorption of bacteria and their effect on aqueous lead adsorption .Journal of Chemical Technology and Biotechnology, 2001, 76(12): 1209-1215.https://doi.org/10.1002/jctb.506 URL [本文引用: 1]摘要 Abstract The adsorption of Escherichia coli on different activated carbons has been studied. The activated carbon samples used have been characterized, determining their surface area, pore size distribution, elemental analysis, mineral matter analysis and pH of the point of zero charge. The adsorption capacity of these carbons increased with their hydrophobicity and macropore volume. The number of bacteria adsorbed on the demineralized activated carbon in a solution of pH value equal to the iso-electric point of the carbon was negligible. However, in the presence of cations the proportions of bacterial cells adsorbed were 87.8% (Fe3+), 54.7% (Ca2+) and 24.8% (Mg2+) respectively. This increase in adsorption capacity in the presence of electrolytes has been explained on the basis of both the reduction in electrostatic free energy and the increase in cell surface hydrophobicity due to the metal bound by some compounds of the cell membrane. When the solution pH was intermediate between the pH values of the point of zero charge of the carbon and bacteria the number of bacteria adsorbed increased due to the attractive interactions between the carbon and bacteria. The adsorption of bacteria on activated carbons decreased the porosity and increased the negative charge density of the latter. Depending on the experimental conditions used, the presence of bacteria can enhance the capacity of activated carbons to adsorb lead. 漏 2001 Society of Chemical Industry
[15]	WARNOCK D D, LEHMANN J, KUYPER T W, RILLIG M C.Mycorrhizal responses to biochar in soil - concepts and mechanisms .Plant and Soil, 2007, 300(1/2): 9-20.https://doi.org/10.1007/s11104-007-9391-5 URL [本文引用: 3]摘要 Experiments suggest that biomass-derived black carbon (biochar) affects microbial populations and soil biogeochemistry. Both biochar and mycorrhizal associations, ubiquitous symbioses in terrestrial ecosystems, are potentially important in various ecosystem services provided by soils, contributing to sustainable plant production, ecosystem restoration, and soil carbon sequestration and hence mitigation of global climate change. As both biochar and mycorrhizal associations are subject to management, understanding and exploiting interactions between them could be advantageous. Here we focus on biochar effects on mycorrhizal associations. After reviewing the experimental evidence for such effects, we critically examine hypotheses pertaining to four mechanisms by which biochar could influence mycorrhizal abundance and/or functioning. These mechanisms are (in decreasing order of currently available evidence supporting them): (a) alteration of soil physico-chemical properties; (b) indirect effects on mycorrhizae through effects on other soil microbes; (c) plant-fungus signaling interference and detoxification of allelochemicals on biochar; and (d) provision of refugia from fungal grazers. We provide a roadmap for research aimed at testing these mechanistic hypotheses.
[16]	KOLB S E, FERMANICH K J, DOMNBUSH M E.Effect of charcoal quantity on microbial biomass and activity in temperate soils .Soil Science Society of America Journal, 2009, 73(4): 1173-1181.https://doi.org/10.2136/sssaj2008.0232 URL [本文引用: 1]摘要 Wildfire-produced charcoal is a common component of soils, affecting a range of important abiotic and biotic soil processes. Our ability to predict the effects of charcoal addition to soil is currently limited, however, by our understanding of how charcoal affects the soil microbial community mediating many of these processes. This study sought to improve our understanding of the relationship b...
[17]	颜永毫, 王丹丹, 郑纪勇. 生物炭对土壤N₂O和CH₄排放影响的研究进展 . 中国农学通报, 2013, 29(8): 140-146.https://doi.org/10.3969/j.issn.1000-6850.2013.08.027 URL [本文引用: 1]摘要 In order to study the pathway and mechanism of biochar affecting soil N2O and CH4 emissions, based on comprehensive evaluation of published researches, the factors, pathway and mechanism that biochar affected soil N2O and CH4 emissions were analyzed, the principles of biochar addition in different types of soil were then proposed. The key problems that should be paid attention to in future studies were pointed out: (1) the characters of these two GHG emissions in soil should be explicited, the biochar type should be chosen according to the local soil conditions. (2) The time and amount of biochar addition should be reasonable. (3) Nowadays because of the differences of biochars and soil types that different scholars used, the research conclusions about the effects of biochar on the soil N2O and CH4 emissions were still controversial. In future, the more clear conclusions about the effects of biochar on the soil N2O and CH4 would be attained till the experiments of returning biochar into field according to the local soil type should be continued after the complement of biochar applying standard. YAN Y H, WANG D D,ZHENG J Y.Advances in effects of biochar on the soil N2O and CH4 emissions .Chinese Agricultural Science Bulletin, 2013, 29(8): 140-146. (in Chinese)https://doi.org/10.3969/j.issn.1000-6850.2013.08.027 URL [本文引用: 1]摘要 In order to study the pathway and mechanism of biochar affecting soil N2O and CH4 emissions, based on comprehensive evaluation of published researches, the factors, pathway and mechanism that biochar affected soil N2O and CH4 emissions were analyzed, the principles of biochar addition in different types of soil were then proposed. The key problems that should be paid attention to in future studies were pointed out: (1) the characters of these two GHG emissions in soil should be explicited, the biochar type should be chosen according to the local soil conditions. (2) The time and amount of biochar addition should be reasonable. (3) Nowadays because of the differences of biochars and soil types that different scholars used, the research conclusions about the effects of biochar on the soil N2O and CH4 emissions were still controversial. In future, the more clear conclusions about the effects of biochar on the soil N2O and CH4 would be attained till the experiments of returning biochar into field according to the local soil type should be continued after the complement of biochar applying standard.
[18]	LEHMANN J, RILL M C, THIES J, MASIELLO C A, HOCKADAY W C, CROWLEY D.Biochar effects on soil biota - A review .Soil Biology and Biochemistry, 2011, 43(9):1812-1836.https://doi.org/10.1016/j.soilbio.2011.04.022 URL Magsci [本文引用: 5]摘要 Soil amendment with biochar is evaluated globally as a means to improve soil fertility and to mitigate climate change. However, the effects of biochar on soil biota have received much less attention than its effects on soil chemical properties. A review of the literature reveals a significant number of early studies on biochar-type materials as soil amendments either for managing pathogens, as inoculant carriers or for manipulative experiments to sorb signaling compounds or toxins. However, no studies exist in the soil biology literature that recognize the observed large variations of biochar physico-chemical properties. This shortcoming has hampered insight into mechanisms by which biochar influences soil microorganisms, fauna and plant roots. Additional factors limiting meaningful interpretation of many datasets are the clearly demonstrated sorption properties that interfere with standard extraction procedures for soil microbial biomass or enzyme assays, and the confounding effects of varying amounts of minerals. In most studies, microbial biomass has been found to increase as a result of biochar additions, with significant changes in microbial community composition and enzyme activities that may explain biogeochemical effects of biochar on element cycles, plant pathogens, and crop growth. Yet, very little is known about the mechanisms through which biochar affects microbial abundance and community composition. The effects of biochar on soil fauna are even less understood than its effects on microorganisms, apart from several notable studies on earthworms. It is clear, however, that sorption phenomena, pH and physical properties of biochars such as pore structure, surface area and mineral matter play important roles in determining how different biochars affect soil biota. Observations on microbial dynamics lead to the conclusion of a possible improved resource use due to co-location of various resources in and around biochars. Sorption and thereby inactivation of growth-inhibiting substances likely plays a role for increased abundance of soil biota. No evidence exists so far for direct negative effects of biochars on plant roots. Occasionally observed decreases in abundance of mycorrhizal fungi are likely caused by concomitant increases in nutrient availability, reducing the need for symbionts. In the short term, the release of a variety of organic molecules from fresh biochar may in some cases be responsible for increases or decreases in abundance and activity of soil biota. A road map for future biochar research must include a systematic appreciation of different biochar-types and basic manipulative experiments that unambiguously identify the interactions between biochar and soil biota.
[19]	O'NEILL B, GROSSMAN J, TSAI M T, GOMES J E, LEHMANN J, PETERSON J, NEVES E, THIES J E. Bacterial community composition in Brazilian Anthrosols and adjacent soils characterized using culturing and molecular identification .Microbial Ecology, 2009, 58(1): 23-35.https://doi.org/10.1109/MIC.2007.108 URLPMID:19381712 Magsci [本文引用: 2]摘要 Microbial community composition was examined in two soil types, Anthrosols and adjacent soils, sampled from three locations in the Brazilian Amazon. The Anthrosols, also known as Amazonian dark earths, are highly fertile soils that are a legacy of pre-Columbian settlement. Both Anthrosols and adjacent soils are derived from the same parent material and subject to the same environmental conditions, including rainfall and temperature; however, the Anthrosols contain high levels of charcoal-like black carbon from which they derive their dark color. The Anthrosols typically have higher cation exchange capacity, higher pH, and higher phosphorus and calcium contents. We used culture media prepared from soil extracts to isolate bacteria unique to the two soil types and then sequenced their 16S rRNA genes to determine their phylogenetic placement. Higher numbers of culturable bacteria, by over two orders of magnitude at the deepest sampling depths, were counted in the Anthrosols. Sequences of bacteria isolated on soil extract media yielded five possible new bacterial families. Also, a higher number of families in the bacteria were represented by isolates from the deeper soil depths in the Anthrosols. Higher bacterial populations and a greater diversity of isolates were found in all of the Anthrosols, to a depth of up to 1 m, compared to adjacent soils located within 50-500 m of their associated Anthrosols. Compared to standard culture media, soil extract media revealed diverse soil microbial populations adapted to the unique biochemistry and physiological ecology of these Anthrosols.
[20]	FIERER N, SCHIMEL J P, HOLDEN P A.Variations in microbial community composition through two soil depth profiles .Soil Biology & Biochemistry, 2003, 35(1): 167-176.https://doi.org/10.1016/S0038-0717(02)00251-1 URL [本文引用: 3]摘要 Soil profiles are often many meters deep, but with the majority of studies in soil microbiology focusing exclusively on the soil surface, we know very little about the nature of the microbial communities inhabiting the deeper soil horizons. We used phospholipid fatty acid (PLFA) analysis to examine the vertical distribution of specific microbial groups and to identify the patterns of microbial abundance and community-level diversity within the soil profile. Samples were collected from the soil surface down to 2 m in depth from two unsaturated Mollisol profiles located near Santa Barbara, CA, USA. While the densities of microorganisms were generally one to two orders of magnitude lower in the deeper horizons of both profiles than at the soil surface, approximately 35% of the total quantity of microbial biomass found in the top 2 m of soil is found below a depth of 25 cm. Principal components analysis of the PLFA signatures indicates that the composition of the soil microbial communities changes significantly with soil depth. The differentiation of microbial communities within the two profiles coincides with an overall decline in microbial diversity. The number of individual PLFAs detected in soil samples decreased by about a third from the soil surface down to 2 m. The ratios of cyclopropyl/monoenoic precursors and total saturated/total monounsaturated fatty acids increased with soil depth, suggesting that the microbes inhabiting the deeper soil horizons are more carbon limited than surface-dwelling microbes. Using PLFAs as biomarkers, we show that Gram-positive bacteria and actinomycetes tended to increase in proportional abundance with increasing soil depth, while the abundances of Gram-negative bacteria, fungi, and protozoa were highest at the soil surface and substantially lower in the subsurface. The vertical distribution of these specific microbial groups can largely be attributed to the decline in carbon availability with soil depth.
[21]	潘全良, 宋涛, 陈坤, 徐晓楠, 彭靖, 战秀梅, 王月, 韩晓日. 连续6年施用生物炭和炭基肥对棕壤生物活性的影响 . 华北农学报, 2016, 31(3): 225-232.https://doi.org/10.7668/hbnxb.2016.03.033 URL Magsci [本文引用: 2]摘要为了明确生物炭与传统土壤培肥方式对土壤中生物活性的影响,通过连续6年微区定位试验,以传统的土壤培肥方式作为对照,探究较长时间施用生物炭和炭基肥对土壤酶活性的影响,以更好地揭示施用生物炭对农业土壤微生态环境的影响,为生物炭农用提供理论参考。定位试验于2009年开始,连续6年进行了花生微区田间试验(2 m2)。试验设4个处理,分别为秸秆还田+NPK(CS)、施用猪厩肥+NPK(PMC)、生物炭+NPK(BIO)和炭基肥(BF)处理。测定了2014年播前以及花生各生育时期土壤中过氧化氢酶、蔗糖酶、脲酶以及微生物量碳和可溶性有机碳含量等指标。结果表明,连续施用6年后,BIO处理中土壤过氧化氢酶活性除播前和成熟期与CS及PMC处理相近外,其他生育时期均显著(P0.05)低于这2个处理;土壤蔗糖酶活性除成熟期外其他时期均高于CS和PMC处理。BF处理中土壤过氧化氢酶活性除播前处于较高水平,其他生育时期与CS和PMC相比均处于较低水平;土壤蔗糖酶活性除结荚期外其他生育时期均接近或者低于CS和PMC处理。与CS和PMC处理相比,BIO和BF处理对脲酶活性的影响没有表现出明显的规律性。BIO和BF处理除成熟期外,其他生育期中土壤微生物量碳含量均低于PMC处理。在花针期相对于CS和PMC处理,BIO处理中可溶性有机碳含量提高,BF中处理可溶性有机碳含量降低。与CS和PMC相比,施用生物炭会抑制土壤过氧化氢酶活性,提高蔗糖酶活性,提高了土壤可溶性有机碳含量;施用炭基肥抑制了过氧化氢酶、蔗糖酶活性,降低了可溶性有机碳含量,生物炭和炭基肥对脲酶活性的影响没有表现出明显规律性,对微生物活性提高的作用在秸秆还田和施用猪厩肥之间。 PAN Q L, SONG T, CHEN K, XU X N, PENG J, ZHAN X M, WANG Y, HAN X R.Influences of 6-year application of biochar and biochar-based compound fertilizer on soil bioactivity on brown soil .Acta Abriculturae Boreali-simica, 2016, 31(3): 225-232. (in Chinese)https://doi.org/10.7668/hbnxb.2016.03.033 URL Magsci [本文引用: 2]摘要为了明确生物炭与传统土壤培肥方式对土壤中生物活性的影响,通过连续6年微区定位试验,以传统的土壤培肥方式作为对照,探究较长时间施用生物炭和炭基肥对土壤酶活性的影响,以更好地揭示施用生物炭对农业土壤微生态环境的影响,为生物炭农用提供理论参考。定位试验于2009年开始,连续6年进行了花生微区田间试验(2 m2)。试验设4个处理,分别为秸秆还田+NPK(CS)、施用猪厩肥+NPK(PMC)、生物炭+NPK(BIO)和炭基肥(BF)处理。测定了2014年播前以及花生各生育时期土壤中过氧化氢酶、蔗糖酶、脲酶以及微生物量碳和可溶性有机碳含量等指标。结果表明,连续施用6年后,BIO处理中土壤过氧化氢酶活性除播前和成熟期与CS及PMC处理相近外,其他生育时期均显著(P0.05)低于这2个处理;土壤蔗糖酶活性除成熟期外其他时期均高于CS和PMC处理。BF处理中土壤过氧化氢酶活性除播前处于较高水平,其他生育时期与CS和PMC相比均处于较低水平;土壤蔗糖酶活性除结荚期外其他生育时期均接近或者低于CS和PMC处理。与CS和PMC处理相比,BIO和BF处理对脲酶活性的影响没有表现出明显的规律性。BIO和BF处理除成熟期外,其他生育期中土壤微生物量碳含量均低于PMC处理。在花针期相对于CS和PMC处理,BIO处理中可溶性有机碳含量提高,BF中处理可溶性有机碳含量降低。与CS和PMC相比,施用生物炭会抑制土壤过氧化氢酶活性,提高蔗糖酶活性,提高了土壤可溶性有机碳含量;施用炭基肥抑制了过氧化氢酶、蔗糖酶活性,降低了可溶性有机碳含量,生物炭和炭基肥对脲酶活性的影响没有表现出明显规律性,对微生物活性提高的作用在秸秆还田和施用猪厩肥之间。
[22]	BOSSIO D A, SCOW K M.Impacts of carbon and flooding on soil microbial communities: Phospholipid fatty acid profiles and substrate utilization patterns .Microbial Ecology, 1998, 35(3): 265-278.URL [本文引用: 1]
[23]	齐鸿雁, 薛凯, 张洪勋. 磷脂脂肪酸谱图分析方法及其在微生物生态学领域的应用 . 生态学报, 2003, 23(8): 1576-1582.URL Magsci [本文引用: 2]摘要应用磷脂脂肪酸谱图分析技术对微生物群落进行定量分布 ,克服了传统的微生物培养方法和显微技术的局限性。介绍了磷脂脂肪酸谱图分析方法及其在微生物生态学领域中的应用 ,包括对微生物群落的生物量、群落结构、营养状况和新陈代谢活动等方面的研究。 QI H Y, XUE K, ZHANG H X.Phospholipid fatty acid analysis and its applications in microbial ecology .Acta Ecologica Sinica, 2003, 23(8): 1576-1582. (in Chinese)URL Magsci [本文引用: 2]摘要应用磷脂脂肪酸谱图分析技术对微生物群落进行定量分布 ,克服了传统的微生物培养方法和显微技术的局限性。介绍了磷脂脂肪酸谱图分析方法及其在微生物生态学领域中的应用 ,包括对微生物群落的生物量、群落结构、营养状况和新陈代谢活动等方面的研究。
[24]	SCHNECKER J R, WILD B, FUCHSLUEGER L, RICHTER A.A field method to store samples from temperate mountain grassland soils for analysis of phospholipid fatty acids .Soil Biology and Biochemistry, 2012, 51(8): 81-83.https://doi.org/10.1016/j.soilbio.2012.03.029 URLPMID:22865936 [本文引用: 2]摘要 The storage of soil samples for PLFA analysis can lead to shifts in the microbial community composition. We show here that conserving samples in RNAlater, which is already widely used to store samples for DNA and RNA analysis, proved to be as sufficient as freezing at02612002°C and preferable over storage at 402°C for temperate mountain grassland soil. The total amount of extracted PLFAs was not changed by any storage treatment. Storage at 402°C led to an alteration of seven out of thirty individual biomarkers, while freezing and storage in RNAlater caused changes in the amount of fungal biomarkers but had no effect on any other microbial group. We therefore suggest that RNAlater could be used to preserve soil samples for PLFA analysis when immediate extraction or freezing of samples is not possible, for example during sampling campaigns in remote areas or during transport and shipping.
[25]	FROSTEG?R A, B??TH E. The use of phospholipid fatty acid analysis to estimate bacterial and fungal biomass in soil .Biology and Fertility of Soils, 1996, 22(1): 59-65.https://doi.org/10.1007/BF00384433 URL [本文引用: 2]摘要 The cell content of 12 bacterial phospholipid fatty acids (PLFA) was determined in bacteria extracted from soil by homogenization/centrifugation. The bacteria were enumerated using acridine orange direct counts. An average of 1.40×10 -17 mol bacterial PLFA cell -1 was found in bacteria extracted from 15 soils covering a wide range of pH and organic matter contents. With this factor, the bacterial biomass based on PLFA analyses of whole soil samples was calculated as 1.0–4.8 mg bacterial C g -1 soil C. The corresponding range based on microscopical counts was 0.3–3.0 mg bacterial C g -1 soil C. The recovery of bacteria from the soils using homogenization/centrifugation was 2.6–16% (mean 8.7%) measured by PLFA analysis, and 12–61% (mean 26%) measured as microscopical counts. The soil content of the PLFA 18:2ω6 was correlated with the ergosterol content ( r =0.92), which supports the use of this PLFA as an indicator of fungal biomass. The ratio 18:2ω6 to bacterial PLFA is therefore suggested as an index of the fungal:bacterial biomass ratio in soil. An advantage with the method based on PLFA analyses is that the same technique and even the same sample is used to determine both fungi and bacteria. The fungal:bacterial biomass ratio calculated in this way was positively correlated with the organic matter content of the soils ( r =0.94).
[26]	OLSSON P A.Signature fatty acids provide tools for determination of the distribution and interactions of mycorrhizal fungi in soil .FEMS Microbiology Ecology, 1999, 29(4): 303-310.https://doi.org/10.1111/j.1574-6941.1999.tb00621.x URL [本文引用: 1]摘要 Abstract Mycorrhizal fungi form extensive mycelia in soil and play significant roles in most soil ecosystems. The estimation of their biomasses is thus of importance in order to understand their possible role in soil nutrient processes. For arbuscular mycorrhizal (AM) fungi the signature fatty acid 16:1蠅5 provides a new and promising tool for the estimation of AM fungal biomass in soil and roots. For ectomycorrhizal fungi 18:2蠅6,9 dominates among the fatty acids and can be used as an indicator of mycelial biomass of these fungi in soil in experimental systems. In biomass estimation primarily the phospholipid fatty acids (PLFAs) are suitable. Through the use of specific PLFAs it is possible to study interactions between mycorrhizal mycelia and bacteria in soil as well as between AM fungal mycelia and mycelia of saprophytic and parasitic fungi in soil and in roots. AM fungi, in particular, store a large proportion of their energy as lipids and by using the signature fatty acids it is possible to determine the relation between membrane and storage lipids, which could be an indication of energy storage levels. Various aspects of how the fatty acid signatures can be used for studies related to questions of biomass distribution and nutritional status of mycorrhizal fungi are discussed.
[27]	HILL G T, MITKOWSKI N A, ALDRICH-WOLFE L, EMELE L R, JURKONIE D D, FICKE A, MALDONADO-RAMIREZ S, LYNCH S T, NELSON E B.Methods for assessing the composition and diversity of soil microbial communities .Applied Soil Ecology, 2000, 15(1): 25-36.https://doi.org/10.1016/S0929-1393(00)00069-X URL [本文引用: 1]摘要 Soil microorganisms play important roles in soil quality and plant productivity. The development of effective methods for studying the diversity, distribution, and behavior of microorganisms in soil habitats is essential for a broader understanding of soil health. Traditionally, the analysis of soil microbial communities has relied on culturing techniques using a variety of culture media designed to maximize the recovery of diverse microbial populations. However, only a small fraction (<0.1%) of the soil microbial community has been accessible with this approach. To overcome these problems, other methods such as the analysis of phospholipid fatty acids and community-level physiological profiles have been utilized in an attempt to access a greater proportion of the soil microbial community. In recent years, molecular methods for soil microbial community analysis have provided a new understanding of the phylogenetic diversity of microbial communities in soil. Among the most useful of these methods are those in which small subunit rRNA genes are amplified from soil-extracted nucleic acids. Using these techniques, it is possible to characterize and study soil microbes that currently cannot be cultured. Microbial rRNA genes can be detected directly from soil samples and sequenced. These sequences can then be compared with those from other known microorganisms. Additionally, group- and taxon-specific oligonucleotide probes can be developed from these sequences making direct visualization of microorganisms in soil habitats possible. The use of these techniques provides new ways of assessing soil microbial diversity and ultimately, a more complete understanding of the potential impacts of environmental processes and human activities on responses of soil microorganisms. Information gained from such studies will have direct impacts on our understanding of the role of microbial processes in soil health.
[28]	SUUTARI M, LAAKSO S.Microbial fatty acids and thermal adaptation .Critical Reviews in Microbiology, 1994, 20(4): 285-328.https://doi.org/10.3109/10408419409113560 URLPMID:7857519 [本文引用: 1]摘要 The existing literature on the role of fatty acids in microbial temperature adaptation is reviewed. Several modes of change of cellular fatty acids at varying environmental temperatures are shown to exist in yeasts and fungi, Gram-negative bacteria, and bacteria containing iso- and anteiso-branched fatty acids, as well as in a few Gram-positive bacteria. Consequently, the degree of fatty acid unsaturation and cyclization, fatty acid chain length, branching, and cellular fatty acid content increase, decrease, or remain unaltered on lowering the temperature. Moreover, microorganisms seem to be able to change from one mode or alter the cellular fatty acid profile temperature dependently to another on lowering the temperature, as well as even within the same growth temperature range, depending on growth conditions. Therefore, the effect of the temperature on cellular fatty acids appears to be more complicated than known earlier. However, similarities found in the modes of change of cellular fatty acids at varying environmental temperatures in several microorganisms within the above mentioned groups support the existence of a limited amount of common regulatory mechanisms. The models presented enable the prediction of temperature-induced changes occurring in the fatty acids of microorganisms, and enzymatic steps of the fatty acid biosynthesis that possibly are under temperature control.
[29]	KIMETU J M, LEHMANN J, KRULL E, SINGH B, JOSEPH S.Stability and stabilisation of biochar and green manure in soil with different organic carbon contents .Australian Journal of Soil Research, 2010, 48(7): 577-585.https://doi.org/10.1071/SR10036 URL [本文引用: 1]摘要 Due to its recalcitrance against microbial degradation, biochar is very stable in soil compared to other organic matter additions, making its application to soils a suitable approach for the build-up of soil organic carbon (SOC). The net effects of such biochar addition also depend on its interactions with existing organic matter in soils. A study was established to investigate how the status of pre-existing soil organic matter influences biochar stabilisation in soil in comparison to labile organic additions. Carbon loss was greater in the C-rich sites (C content 58.065g65C/kg) than C-poor soils (C content 21.0–24.065g65C/kg), regardless of the quality of the applied organic resource. Biochar-applied, C-rich soil showed greater C losses, by >0.565kg/m2.year, than biochar-applied C-poor soil, whereas the difference was only 0.165kg/m2.year with Tithonia diversifolia green manure. Biochar application reduced the rate of CO2-C loss by 27%, and T. diversifolia increased CO2-C losses by 22% in the C-poor soils. With biochar application, a greater proportion of C (6.8 times) was found in the intra-aggregate fraction per unit C respired than with green manure, indicating a more efficient stabilisation in addition to the chemical recalcitrance of biochar. In SOC-poor soils, biochar application enriched aromatic-C, carboxyl-C, and traces of ketones and esters mainly in unprotected organic matter and within aggregates, as determined by Fourier-transform infrared spectroscopy. In contrast, additions of T. diversifolia biomass enriched conjugated carbonyl-C such as ketones and quinones, as well as CH deformations of aliphatic-C mainly in the intra-aggregate fraction. The data indicate that not only the stability but also the stabilisation of biochar exceeds that of a labile organic matter addition such as green manure.
[30]	BAGREEV A, BANDOSZ T J, LOCKE D C.Pore structure and surface chemistry of adsorbents obtained by pyrolysis of sewage sludge-derived fertilizer .Carbon, 2001, 39(13): 1971-1979.https://doi.org/10.1016/S0008-6223(01)00026-4 URL Magsci [本文引用: 1]摘要 Terrene, a fertilizer product derived from New York City municipal sewage sludge, was pyrolyzed at various temperatures between 400 and 950°C. The pore structure and surface chemistry of the adsorbent materials obtained were characterized using nitrogen adsorption, thermal analysis, potentiometric titration and FTIR. The adsorbents contain a high percentage of inorganic matter (up to 70%) and only up to 30% carbon. The results show that microporosity is developed within the carbon deposit and at the organic/inorganic interface with increasing temperature of heat treatment. An increase in the pyrolysis temperature also results in significant changes in the surface chemistry towards the development of basic nitrogen centers. In particular there was an increase in the pH values of adsorbents’ surfaces from 7 to 11.
[31]	CAO X, HARRIS W.Properties of dairy-manure-derived biochar pertinent to its potential use in remediation .Bioresource Technology, 2010, 101(14): 5222-5228.https://doi.org/10.1016/j.biortech.2010.02.052 URLPMID:20206509 [本文引用: 1]摘要 Conversion of waste products into biochar (BC) is being considered as one of several waste disposal and recycling options. In this study, we produced BC from dairy manures by heating at low temperatures (81500 °C) and under abundant air condition. The resultant BC was characterized for physical, chemical, and mineralogical properties specifically related to its potential use in remediation. The BC from all manures behaved similarly. Surface area, ash content, and pH of the BC increased as temperature increased, while yield decreased with increasing temperature. The BC was rich in mineral elements such as N, Ca, Mg, and P in addition to C, and concentrations of C and N decreased with increasing temperature as a result of combustion and volatilization; while P, Ca, and Mg increased as temperature increased. For example, C significantly decreased from 36.8% at 100 °C to 1.67% at 500 °C; whereas P increased from 0.91% to 2.66%. Water soluble P, Ca, and Mg increased when heated to 200 °C but decreased at higher temperatures likely due to increased crystallization of Ca–Mg–P, as supported by the formation of whitlockite (Ca,Mg) 3(PO 4) 2 following 500 °C treatment. The presence of whitlockite was evidenced by X-ray diffraction analysis. Quartz and calcite were present in all BC produced. The BC showed appreciable capability of adsorption for Pb and atrazine from aqueous solution, with Pb and atrazine removal by as high as 100% and 77%, respectively. The results indicated that dairy manure can be converted into biochar as an effective adsorbent for application in environmental remediation.
[32]	SCHMIDT M W I, NOACK A G. Black carbon in soils and sediments: Analysis, distribution, implications, and current challenges .Global Biogeochemical Cycles, 2000, 14(3): 777-793.https://doi.org/10.1029/1999GB001208 URL [本文引用: 1]摘要 This review highlights the ubiquity of black carbon (BC) produced by incomplete combustion of plant material and fossil fuels in peats, soils, and lacustrine and marine sediments. We examine various definitions and analytical approaches and seek to provide a common language. BC represents a continuum from partly charred material to graphite and soot particles, with no general agreement on clear-cut boundaries. Formation of BC can occur in two fundamentally different ways. Volatiles recondense to highly graphitized soot-BC, whereas the solid residues form char-BC. Both forms of BC are relatively inert and are distributed globally by water and wind via fluvial and atmospheric transport. We summarize, chronologically, the ubiquity of BC in soils and sediments since Devonian times, differentiating between BC from vegetation fires and from fossil fuel combustion. BC has important implications for various biological, geochemical and environmental processes. As examples, BC may represent a significant sink in the global carbon cycle, affect the Earth's radiative heat balance, be a useful tracer for Earth's fire history, build up a significant fraction of carbon buried in soils and sediments, and carry organic pollutants. On land, BC seems to be abundant in dark-colored soils, affected by frequent vegetation burning and fossil fuel combustion, thus probably contributing to the highly stable aromatic components of soil organic matter. We discuss challenges for future research. Despite the great importance of BC, only limited progress has been made in calibrating analytical techniques. Progress in the quantification of BC is likely to come from systematic intercomparison using BCs from different sources and in different natural matrices. BC identification could benefit from isotopic and spectroscopic techniques applied at the bulk and molecular levels. The key to estimating BC stocks in soils and sediments is an understanding of the processes involved in BC degradation on a molecular level. A promising approach would be the combination of short-term laboratory experiments and long-term field trials.
[33]	YAO H, HE Z, WILSON M J, CAMPBELL C D.Microbial biomass and community structure in a sequence of soils with increasing fertility and changing land use .Microbial Ecology, 2000, 40(3): 223-237.https://doi.org/10.1007/s002480000053 URL Magsci [本文引用: 1]摘要 <a name="Abs1"></a><div class="AbstractPara"> <div class="">The microbial biomass and community structure of eight Chinese red soils with different fertility and land use history was investigated. Two community based microbiological measurements, namely, community level physiological profiling (CLPP) using Biolog sole C source utilization tests and phospholipid fatty acid (PLFA) profiles, were used to investigate the microbial ecology of these soils and to determine how land use alters microbial community structure. Microbial biomass-C and total PLFAs were closely correlated to organic carbon and total nitrogen, indicating that these soil microbial measures are potentially good indices of soil fertility in these highly weathered soils. Metabolic quotients and C source utilization were not correlated with organic carbon or microbial biomass. Multivariate analysis of sole carbon source utilization patterns and PLFAs demonstrated that land use history and plant cover type had a significant impact on microbial community structure. PLFAs showed these differences more than CLPP methods. Consequently, PLFA analysis was a better method for assessing broad-spectrum community differences and at the same time attempting to correlate changes with soil fertility. Soils from tea orchards were particularly distinctive in their CLPP. A modified CLPP method, using absorbance readings at 405 nm and different culture media at pH values of 4.7 and 7.0, showed that the discrimination obtained can be influenced by the culture conditions. This method was used to show that the distinctive microbial community structure in tea orchard soils was not, however, due to differences in pH alone.
[34]	MARSCHNER P, KANDELER E, MARSCHNER B.Structure and function of the soil microbial community in a long-term fertilizer experiment .Soil Biology and Biochemistry, 2003, 35(3): 453-461.https://doi.org/10.1016/S0038-0717(02)00297-3 URL [本文引用: 1]摘要 The effect of organic and inorganic fertiliser amendments is often studied shortly after addition of a single dose to the soil but less is known about the long-term effects of amendments. We conducted a study to determine the effects of long-term addition of organic and inorganic fertiliser amendments at low rates on soil chemical and biological properties. Surface soil samples were taken from an experimental field site near Cologne, Germany in summer 2000. At this site, five different treatments were established in 1969: mineral fertiliser (NPK), crop residues removed (mineral only); mineral fertiliser with crop residues; manure 5.2 t ha 611 yr 611; sewage sludge 7.6 t ha 611 yr 611 or straw 4.0 t ha 611 yr 611 with 10 kg N as CaCN 2 t straw 611. The organic amendments increased the C org content of the soil but had no significant effect on the dissolved organic C (DOC) content. The C/N ratio was highest in the straw treatment and lowest in the mineral only treatment. Of the enzymes studied, only protease activity was affected by the different amendments. It was highest after sewage amendment and lowest in the mineral only treatment. The ratios of Gram+ to Gram61 bacteria and of bacteria to fungi, as determined by signature phospholipid fatty acids, were higher in the organic treatments than in the inorganic treatments. The community structure of bacteria and eukaryotic microorganisms was assessed by denaturing gradient gel electrophoresis (DGGE) and redundancy discriminate analyses of the DGGE banding patterns. While the bacterial community structure was affected by the treatments this was not the case for the eukaryotes. Bacterial and eukaryotic community structures were significantly affected by C org content and C/N ratio.
[35]	INGWERSEN J, POLL C, STRECK T, KANDELER E.Micro-scale modelling of carbon turnover driven by microbial succession at a biogeochemical interface .Soil Biology and Biochemistry, 2008, 40(4): 864-878.https://doi.org/10.1016/j.soilbio.2007.10.018 URL Magsci [本文引用: 1]摘要 The detritusphere is a very thin but microbiological highly active zone in soil. To trace the fate of litter carbon in the detritusphere we developed a new 1D dynamic mechanistic model. In a microcosm experiment soil cores were incubated with C labelled rye residues (C=299鈥), which were placed on the surface. Microcosms were sampled after 3, 7, 14, 28, 56 and 84 days and soil cores were separated into layers of increasing distance to the litter. Gradients in soil organic carbon (TOC), dissolved organic carbon (DOC), microbial biomass and activity were detected over a distance of 3聽mm from the litter layer. The newly developed 1D model simulates both the total carbon and the C carbon pools and fluxes, so that it was possible to include the C data in model optimisation. The special feature of the model is that it operates with two decomposer populations; the first one is assumed to be dominated by bacteria (initial-stage decomposer) and second one by fungi (late-stage decomposer). Moreover, in the model the DOC pool is divided into two sub pools. Each DOC pool is consumed by one of the decomposer populations. After parameter optimisation the model was well suited to simulate the experimental data. The model explained 92% of the observed variance. The model output provides a comprehensive insight into the carbon cycling within the detritusphere. The simulation results showed among others that after 84 days about 10% of total litter C was transferred to the soil organic matter (SOM) pool. Only 3% was located in the microbial biomass. From the evolved CO 71% was litter-derived and 29% was soil-derived. From the litter-derived CO, 69% was directly formed in the litter layer. The remaining 31% was transported to soil before mineralisation. Our study shows that a combination of experimental work and mathematical modelling is a powerful approach to provide a comprehensive insight into the small-scale carbon turnover in soil.
[36]	JIN H.Characterization of microblal life colonizing biochar and biochar-amended soils . Ithaca: Cornell University, 2010. [本文引用: 1]
[37]	李新, 焦燕, 杨铭德. 用磷脂脂肪酸(plfa)谱图技术分析内蒙古河套灌区不同盐碱程度土壤微生物群落多样性 . 生态科学, 2014, 33(3): 488-494.URL Magsci [本文引用: 1]摘要应用磷脂脂肪酸(PLFA)法分析了内蒙古河套灌区3种不同盐碱程度(盐土、强度盐化土、轻度盐化土)土壤细菌、真菌和原生动物等微生物多样性。结果表明: 盐土土壤微生物的PLFA总量显著低于强度盐化土和轻度盐化土; 三种不同盐碱程度土壤中的微生物均以细菌为主,  盐土的细菌PLFA含量较强度盐化土和轻度盐化土的细菌PLF含量都显著降低; 以27 种PLFA含量为样本进行聚类分析, 发现土壤盐碱化程度不同,  土壤微生物结构必然发生变化; Shannon- Wiener等多样性指数分析可得盐碱程度越大, 主要土壤微生物PLFA标记物多样性越单一, 反之则越丰富; 以PLFA标记物为物种, 以土壤含盐量、pH、土壤有机质、土壤全氮和土壤全磷为环境变量, 借助CANOCO软件主分分析生成物种-环境双序图, 两个排序轴对物种变量的解释量达94.3%, 土壤含盐量、pH与第一主成分轴呈正相关,  相关系数分别为0.8757, 0.9091; 土壤有机质与土壤全氮与第一主成分轴呈负相关,  相关系数分别为–0.9398和–0.8992。 LI X, JIAO Y, YANG M D.Microbial diversity of different saline-alkaline soil analyzing by PLFA in the Hetao area of inner Mongolia . Ecological Science, 2014, 33(3): 488-494. (in Chinese)URL Magsci [本文引用: 1]摘要应用磷脂脂肪酸(PLFA)法分析了内蒙古河套灌区3种不同盐碱程度(盐土、强度盐化土、轻度盐化土)土壤细菌、真菌和原生动物等微生物多样性。结果表明: 盐土土壤微生物的PLFA总量显著低于强度盐化土和轻度盐化土; 三种不同盐碱程度土壤中的微生物均以细菌为主,  盐土的细菌PLFA含量较强度盐化土和轻度盐化土的细菌PLF含量都显著降低; 以27 种PLFA含量为样本进行聚类分析, 发现土壤盐碱化程度不同,  土壤微生物结构必然发生变化; Shannon- Wiener等多样性指数分析可得盐碱程度越大, 主要土壤微生物PLFA标记物多样性越单一, 反之则越丰富; 以PLFA标记物为物种, 以土壤含盐量、pH、土壤有机质、土壤全氮和土壤全磷为环境变量, 借助CANOCO软件主分分析生成物种-环境双序图, 两个排序轴对物种变量的解释量达94.3%, 土壤含盐量、pH与第一主成分轴呈正相关,  相关系数分别为0.8757, 0.9091; 土壤有机质与土壤全氮与第一主成分轴呈负相关,  相关系数分别为–0.9398和–0.8992。
[38]	FTDE V, HOFFLAND E, NVAN E, BRUSSAARD L, BLOEM J.Fungal/bacterial ratios in grasslands with contrasting nitrogen management .Soil Biology and Biochemistry, 2006, 38(8): 2092-2103.https://doi.org/10.1016/j.soilbio.2006.01.008 URL [本文引用: 2]摘要 It is frequently hypothesised that high soil fungal/bacterial ratios are indicative for more sustainable agricultural systems. Increased F / B ratios have been reported in extensively managed grasslands. To determine the shifts in fungal/bacterial biomass ratio as influenced by grassland management and to find relations with nitrogen leaching potential, we sampled a two-year-old field experiment at an organic experimental farm in the eastern part of The Netherlands. The effect of crop (grass and grass-clover), N application rate (0, 40, 80, 120 kg N ha - 1) and manure type (no manure, farm yard manure and slurry) on the F / B ratio within three growing seasons was tested, as well as relations with soil and crop characteristics, nitrate leaching and partial N balance. Biomass of fungi and bacteria was calculated after direct counts using epifluorescence microscopy. Fungal and bacterial biomass and the F / B ratio were higher in grass than in grass-clover. The F / B ratio decreased with increasing N application rate and multiple regression analysis revealed a negative relationship with pH. Bacterial activity (measured as incorporation of [ 3H]thymidine and [ 14C]leucine into bacterial DNA and proteins) showed the exact opposite: an increase with N application rate and pH. Leaching increased with N application rate and was higher in grass-clover than in grass. Partial N balance was more positive at a higher N application rate and showed an inverse relationship with fungal biomass and F / B ratio. We conclude that the fungal/bacterial biomass ratio quickly responded to changes in management. Grasslands with higher N input showed lower F / B ratios. Grass-clover had a smaller fungal biomass and higher N leaching than grass. In general, a higher fungal biomass indicated a lower nitrogen leaching and a more negative partial N balance (or smaller N surplus), but more observations are needed to confirm the relationship between F / B ratio and sustainability.
[39]	FROSTEG?RD ?, B??THE, TUNLIO A. Shifts in the structure of soil microbial communities in limed forests as revealed by phospholipid fatty acid analysis .Soil Biology and Biochemistry, 1993, 25(6): 723-730.https://doi.org/10.1016/0038-0717(93)90113-P URL [本文引用: 1]摘要 The effects of lime and wood-ash on the microbial community structure were evaluated by analyzing the phospholipid fatty acid (PLFA) composition of soils from two areas in the south of Sweden. A pine forest was amended with lime or ash at two concentrations, and a spruce forest was limed at one concentration. The treatments were carried out 5–6 years before sampling and raised the pH from approx. 4.0 to values between 4.8 and 7.0. At both sites there was a difference in the PLFA composition between the treated plots and the controls. The changes found were similar at both sites and correlated to the pH changes. No difference was found between limed plots and those treated with wood-ash. The methyl-branched fatty acids i15:0, i16:0 and 10Me16:0, the monounsaturated fatty acids 16: 1ω 7t and 18: 1ω 9, the cyclopropane fatty acid cy 19:0, and the saturated fatty acid 20:0 were more abundant in the control plots. In the plots with the highest pH there was a three-fold increase in the fatty acid 16: lω 5. An increase was also found for the fatty acids i14:0, 16:lω9, 16:lω 7c, cy17:0, 18:lω 7 and 10Me18:0. No effect on 18:2ω6 was found. The changes in PLFA pattern indicated that the increased pH caused a shift in the bacterial community to more Gram-negative and fewer Gram-positive bacteria, while the amount of fungi was unaffected. The increase in 10Me18:0 in limed soils indicated an increase in actinomycetes.
[40]	MCKINLEY V L, PEACOCK A D, WHITE D C.Microbial community PLFA and PHB responses to ecosystem restoration in tallgrass prairie soils .Soil Biology and Biochemistry, 2005, 37(10): 1946-1958.https://doi.org/10.1016/j.soilbio.2005.02.033 URL [本文引用: 1]摘要 Native North American prairie grasslands are renowned for the richness of their soils, having excellent soil structure and very high organic content and microbial biomass. In this study, surface soils from three prairie restorations of varying ages and plant community compositions were compared with a nearby undisturbed native prairie remnant and a cropped agricultural field in terms of soil physical, chemical and microbial properties. Soil moisture, organic matter, total carbon, total nitrogen, total sulfur, C:N, water-holding capacity and microbial biomass (total PLFA) were significantly greater ( p<0.05) in the virgin prairie remnant as well as the two long-term (21 and 24 year) prairie restorations, compared with the agricultural field and the restoration that was begun more recently (7 years prior to sampling). Soil bulk density was significantly greater ( p<0.05) in the agricultural and recently restored sites. In most cases, the soil quality indicators and microbial community structures in the restoration sites were intermediate between those of the virgin prairie and the agricultural sites. Levels of poly-尾-hydroxybutyrate (PHB) and PLFA indicators of nutritional stress were significantly greater ( p<0.05) in the agricultural and recent restoration sites than in the long-term restorations or the native prairie. Samples could be assigned to the correct site by discriminant analysis of the PLFA data, with the exception that the two long-term restoration sites overlapped. Redundancy analysis showed that prairie age ( p<0.005) was the most important environmental factor in determining the PLFA microbial community composition, with C:N ( p<0.015) also being significant. These findings demonstrate that prairie restorations can lead to improved quality of surface soils. We predict that the conversion of farmland into prairie will shift the soil quality, microbial community biomass and microbial community composition in the direction of native prairies, but with the restoration methods tested it may take many decades to approach the levels found in a virgin prairie throughout the soil profile.
[41]	MARSCHNER P, YANG C H, LIEBEREI R, CROWLEY D E.Soil and plant specific effects on bacterial community composition in the rhizosphere .Soil Biology and Biochemistry, 2001, 33(11): 1437-1445.https://doi.org/10.1016/S0038-0717(01)00052-9 URL [本文引用: 1]摘要 Eubacterial community structures in the plant rhizosphere were examined with respect to plant species, soil type, and root zone location. Three plant species (chickpea, rape and Sudan grass) were grown in intact cores of three California soils (a sandy soil, a sandy loam, and a clay) and were provided with a complete fertilizer solution with or without nitrogen supplied as ammonium nitrate. After 7.5 weeks, the plants were harvested and DNA was extracted from soil adhering to the root tips and from mature root zones at the sites of lateral root emergence. Eubacterial community structures were examined by PCR-DGGE of 16S rDNA to determine the relative abundance and species diversity. While both soil type and nitrogen fertilization affected plant growth, canonical correspondence analyses showed that nitrogen had no significant effect on eubacterial community structures. Eubacterial species diversity was higher in the mature root zones than at the root tips in the sandy soil and the clay but not in the loamy sand. Monte Carlo permutation tests indicated that plant species, root zone and soil type as well as the interactions between these variables had significant effects on community structure. The bacterial rhizosphere community of chickpea was influenced primarily by soil type, whereas root zone was less important. In contrast to chickpea, the community in the rhizosphere of rape and Sudan grass was more affected by the root zone than the soil type. In the sandy soil and the loamy sand, the eubacterial rhizosphere community structure was more affected by the root zone than the plant species and the three plant species had distinct communities. In the clay however, the root zone was less important than the plant species and the rhizosphere communities of chickpea differed from those of rape and Sudan grass. It is concluded that the bacterial community composition in the rhizosphere is affected by a complex interaction between soil type, plant species and root zone location.
[42]	岳琳艳, 郑俊强, 韩士杰, 杨建华, 耿世聪, 陈志杰, 张雪, 谷越. 长白山温带森林不同演替阶段土壤化学性质及微生物群落结构的变化 . 生态学杂志, 2015, 34(9): 2590-2597.URL Magsci [本文引用: 2]摘要土壤微生物是生态系统物质循环和能量流动的驱动者,其群落结构可以用来表征土壤生态过程及其对地上植被变化的响应机制.本研究采用时空替代法在长白山地区选取了阔叶红松林演替序列的5个不同阶段:杨桦幼龄林、杨桦中龄林、杨桦成熟林、阔叶红松成熟林和阔叶红松过熟林,采用磷脂脂肪酸法(PLFA)测定了土壤微生物群落组成和结构,分析了其随地上植被演替过程的变化,同时比较了不同演替阶段土壤化学性质差异. 结果表明:随着演替的正向进行,土壤总有机碳、全碳、全氮、全磷含量显著提高,碳氮比逐渐下降.土壤微生物生物量、群落结构及组成发生明显变化:土壤微生物总PLFAs、细菌PLFAs、革兰氏阳性菌PLFAs、革兰氏阴性菌PLFAs含量显著增加;真菌PLFAs(18:2ω6c)先增加后减少,中期阶段的杨桦成熟林土壤真菌PLFA含量最高,同时细菌/真菌最小;革兰氏阳性菌/革兰氏阴性菌(G+/G-)随着演替的进行逐渐增大.土壤微生物生物量与土壤全碳、总有机碳、全氮、全磷含量呈显著正相关,与碳氮比呈显著负相关;冗余分析(RDA)结果显示,全碳、总有机碳、全氮和碳氮比是影响土壤微生物群落结构的主要因素.本研究表明,随着植被演替的正向进行,土壤质量逐渐提高;土壤微生物群落组成明显改变;土壤微生物群落结构与土壤理化性质显著相关. YUE L Y, ZHENG J Q, HAN S J, YANG J H, GENG S C, CHEN Z J, ZHANG X, GU Y.Soil chemical properties and microbial community structure at different succession stages of temperate forest in Changbai Mountains .Chinese Journal of Ecology, 2015, 34(9): 2590-2597. (in Chinese)URL Magsci [本文引用: 2]摘要土壤微生物是生态系统物质循环和能量流动的驱动者,其群落结构可以用来表征土壤生态过程及其对地上植被变化的响应机制.本研究采用时空替代法在长白山地区选取了阔叶红松林演替序列的5个不同阶段:杨桦幼龄林、杨桦中龄林、杨桦成熟林、阔叶红松成熟林和阔叶红松过熟林,采用磷脂脂肪酸法(PLFA)测定了土壤微生物群落组成和结构,分析了其随地上植被演替过程的变化,同时比较了不同演替阶段土壤化学性质差异. 结果表明:随着演替的正向进行,土壤总有机碳、全碳、全氮、全磷含量显著提高,碳氮比逐渐下降.土壤微生物生物量、群落结构及组成发生明显变化:土壤微生物总PLFAs、细菌PLFAs、革兰氏阳性菌PLFAs、革兰氏阴性菌PLFAs含量显著增加;真菌PLFAs(18:2ω6c)先增加后减少,中期阶段的杨桦成熟林土壤真菌PLFA含量最高,同时细菌/真菌最小;革兰氏阳性菌/革兰氏阴性菌(G+/G-)随着演替的进行逐渐增大.土壤微生物生物量与土壤全碳、总有机碳、全氮、全磷含量呈显著正相关,与碳氮比呈显著负相关;冗余分析(RDA)结果显示,全碳、总有机碳、全氮和碳氮比是影响土壤微生物群落结构的主要因素.本研究表明,随着植被演替的正向进行,土壤质量逐渐提高;土壤微生物群落组成明显改变;土壤微生物群落结构与土壤理化性质显著相关.