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

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。     

积雪覆盖变化对大兴安岭多年冻土区土壤微生物群落结构的影响

高纬度多年冻土区是全球变化的敏感区域,揭示不同雪被覆盖条件下土壤微生物群落结构的演变规律,对于预测寒区森林生态系统对全球变化的响应具有重要意义。以大兴安岭多年冻土区白桦次生林为研究对象,采用磷脂脂肪酸法(PLFA)对比分析自然积雪和遮雪处理土壤微生物群落结构的动态变化特征。结果表明:土壤总磷脂脂肪酸含量在植被生长季初期最高,积雪稳定期最低,其中含量较高的PLFA为18:2ω6,9c、a15:0、i16:0、17:1ω8c、18:1ω9c和16:1ω5c,不同时期各优势PLFA含量存在一定差异。遮雪显著降低积雪稳定期细菌PLFA含量、增加真菌PLFA含量(P0.05),但对其他时期土壤微生物群落结构和多样性均未产生显著影响。冗余分析(RDA)表明,土壤总磷脂脂肪酸、真菌、真菌/细菌和革兰氏阴性菌与土壤含水量、pH和铵态氮均呈显著正相关,细菌和革兰氏阳性菌受土壤总磷、总氮和硝态氮影响较大。     

不同种植年限对花椒根际土壤理化性质和微生物群落的影响

为揭示不同种植年限对花椒根际微生态系统的影响,利用磷脂脂肪酸（PLFAs）生物标记法结合土壤养分和酶活性分析,研究了种植年限1、15、30 a的花椒根际土壤的理化性质和微生物群落变化。结果显示,随着种植年限的增加,土壤pH和多酚氧化酶活性呈下降趋势,土壤速效钾和速效磷在15、30 a时显著增加。土壤微生物总PLFA、真菌、革兰氏阴性细菌（G^-）PLFA含量均随种植年限呈增加趋势,30a时达到最大,分别为401.95、88.14、118.61 μg·g^-1。革兰氏阳性细菌（G ⁺ ）和放线菌PLFA含量随种植年限呈先增后降趋势。细菌/真菌和G⁺/G^-随种植年限增加呈下降趋势,与1 a相比,30 a时分别下降34.14%、38.87%。土壤pH与总PLFA、G^-、细菌、真菌呈显著负相关,与G ⁺ /G^-呈显著正相关;多酚氧化酶活性与真菌呈显著负相关,与细菌/真菌、G ⁺ /G^-呈显著正相关。本研究表明：花椒根际理化性质、酶活性和微生物群落结构随种植年限增加发生了显著变化,根际土壤pH、速效磷、速效钾、多酚氧化酶的变化是影响土壤微生物群落与组成的主要环境因子,真菌和G^-的增加及多酚氧化酶活性的降低可能是诱导花椒根腐病发生的重要原因。     

施磷对干旱胁迫下箭竹根际土壤养分及微生物群落的影响

以箭竹及其根际土壤作为研究对象,采用两因素随机区组实验,设置2种水分处理（正常浇水和干旱胁迫）和2种施磷量处理（施磷和不施磷）,探究施磷对干旱胁迫下箭竹根际土壤养分及微生物群落结构和多样性的影响。结果表明：（1）干旱胁迫显著降低了箭竹根际土壤中微生物量碳、可溶性有机氮和有效磷的含量,虽对箭竹根际土壤微生物群落的多样性无显著影响,但显著降低了箭竹根际土壤中总PLFA（phospholipid fatty acid contents）的含量和真菌、细菌、革兰氏阳性菌与革兰氏阴性菌的PLFA含量以及革兰氏阳性菌/革兰氏阴性菌的PLFA比值,显著改变了箭竹根际土壤微生物群落结构,结果显著降低了箭竹的生物量。（2）施磷显著增加了受旱箭竹根际土壤中微生物量碳和有效磷的含量,虽大体上对受旱箭竹根际土壤微生物群落的多样性无显著影响,但显著增加了受旱箭竹根际土壤中总PLFA和真菌PLFA的含量,并在一定程度上增加了细菌、革兰氏阳性菌、革兰氏阴性菌和放线菌的PLFA含量以及革兰氏阳性菌/革兰氏阴性菌和真菌/细菌的PLFA比值,也在一定程度上改善了受旱箭竹根际土壤微生物群落结构,从而改善受旱箭竹的生长。（3）主成分分析表明,干旱对箭竹根际土壤微生物群落结构的影响显著,而施磷的影响不明显。（4）相关分析发现,箭竹根际土壤微生物群落结构与箭竹根际土壤微生物量碳、可溶性有机氮及箭竹生物量呈显著正相关。综上,干旱降低了箭竹根际土壤养分含量和微生物生物量,改变了箭竹根际土壤微生物群落结构,抑制了箭竹的生长;施磷能增加受旱箭竹根际土壤养分含量和微生物生物量,改善受旱箭竹根际土壤微生物群落结构,进而改善受旱箭竹的生长。     

海拔对辽东栎林地土壤微生物群落的影响

以北京东灵山辽东栎林地土壤为对象,运用氯仿熏蒸-浸提法及磷脂脂肪酸分析(PLFA)法,研究林木生长季节土壤微生物群落随海拔梯度的变化特征.结果表明:随着海拔升高,辽东栎林土壤微生物生物量碳、氮,以及微生物各类群含量均有差异但不显著;土壤细菌/真菌升高,而革兰氏阳性菌(G+)/革兰氏阴性菌(G-)降低.土壤微生物生物量碳、氮以及细菌、真菌、G+细菌、G-细菌的含量与土壤含水量、有机碳、全氮呈显著正相关,土壤真菌含量与土壤碳氮比值呈正相关.土壤微生物群落组成结构(细菌/真菌和G+细菌/G-细菌)的变化主要受土壤温度和土壤含水量的显著影响,说明土壤微生物群落结构对环境条件的变化敏感.随着全球变暖的加剧,暖温带辽东栎林地土壤真菌和G+细菌的比例有升高的趋势.     

高寒草甸连续围封与施肥对土壤微生物群落结构的影响

以放牧为对照,应用PLFA法分析研究了放牧、连续6年围封及围封内连续6年施肥后高寒草甸土壤微生物群落结构的变化.结果表明:围封和围封内施肥对不同土层各菌群和微生物总量均有显著影响,其对0 ～ 10 cm土层微生物的影响大于10～20 cm土层,不同土层的PLFA种类发生显著变化.围封和围封内施肥处理不同土层的革兰氏阴性菌(G-)含量均低于放牧;放牧0 ～10 cm土层中细菌、真菌、革兰氏阳性菌(G+)、微生物总量大干围封和围封内施肥处理,但其放线菌生物量均低于围封和围封内施肥处理;在10～20 cm土层中,各样地土壤中的G+无显著差异,围封土壤中的细菌、真菌、放线菌、微生物总量显著高于放牧,而围封内施肥后各菌群生物量及微生物总量明显下降.围封和围封内施肥不同土层的细菌/真菌均高于放牧;一般饱和脂肪酸/单烯不饱和脂肪酸(SAT/MONO)和革兰氏阳性菌/革兰氏阴性菌(G+/G－),围封处理均低于放牧,围封内施肥处理均高于放牧.连续围封和围封内施肥后降低了土壤微生物活性和土壤生态系统的稳定性.     

基于PLFA指纹图谱表征浓香型酒糟醅微生物群落结构

以9株白酒酿造过程中常见的菌株为对象,研究了不同种属菌株的细胞膜特征组分磷酸脂肪酸(PLFA)的特征,以及检出量与菌株生物量之间的关系.结果表明:供试细菌、放线菌、霉菌和酵母菌的PLFA指纹图谱存在显著差异,各菌株的PLFA指纹图谱可作为区别种属的依据.不同供试菌株生物量在一定范围内与检出的总PLFA量或16:0含量呈线性关系.将不同生物量的革兰氏阳性菌G+、革兰氏阴性菌G-和真菌分别加入糟醅后,检出的PLFA相对含量与对照差异显著.基于PLFA的指纹图谱能够定量或半定量地表征糟醅微生物群落结构特征及动态变化.经对多家酿酒企业糟醅PLFA组成的检测及微生物群落结构的剖析,该方法具有普适性.     

喀斯特地区三种人工林土壤微生物群落结构特征

为揭示不同人工植被修复模式对喀斯特土壤微生物群落的影响,采用氯仿熏蒸提取法和磷脂脂肪酸(phospholipid fatty acid, PLFA)法研究人工构建的降香黄檀(Dalbergia odorifera)纯林(PDOP)、顶果木(Acrocarpus fraxinifolius)纯林(PAFP)、顶果木×降香黄檀混交林(MADP)对土壤微生物生物量及土壤微生物群落结构的影响。结果表明:(1)PDOP的土壤微生物生物量碳(MBC)和微生物生物量氮(MBN)含量显著高于PAFP和MADP,PAFP显著高于MADP。(2)三种人工林土壤真菌、丛枝菌根真菌和总PLFA含量无显著差异,但PDOP土壤细菌、放线菌、丛枝菌根真菌和总PLFA含量均高于PAFP和MADP,PAFP高于MADP。PDOP的土壤细菌、革兰氏阳性菌、革兰氏阴性菌、放线菌的PLFA含量显著高于MADP。MADP的真菌细菌比显著高于PDOP,但与PAFP无显著差异。(3)冗余分析表明,土壤阳离子交换量、pH和C:N是影响土壤微生物群落组成的最主要影响因子。从三种人工林的土壤微生物生物量及微生物群落结构来看,在喀斯特地区MADP并未显示出酸性土地区混交林提高土壤微生物生物量、改善土壤微生物群落结构的优势,但混交林的真菌细菌比最高,更有利于提高土壤生态系统的稳定性。     

增温对长白山苔原土壤微生物群落结构的影响

研究土壤微生物群落结构对温度升高的响应,对预测气候变化条件下土壤微生物以及土壤养分循环具有重要意义。采用开顶箱(OTC,Open-top chamber)增温方法对长白山苔原土壤进行连续两个生长季(6—9月)增温处理,结果表明:增温使土壤磷脂脂肪酸(PLFA,Phospholipid fatty acid)总量降低了16.1%,革兰氏阳性菌/革兰氏阴性菌比值(G+/G-)升高21.2%。增温与对照条件下的G+、G-、细菌、真菌的PLFAs相对含量和真菌/细菌比值在统计上无显著差异,除真菌与G-外,其它指标均存在明显的季节波动。增温与对照条件下,细菌、G+、G+/G-和PLFA总量在土壤温度较高的7、8月份较温度较低的9月份高,真菌/细菌比值则在9月份温度较低时达到最大值。主成分分析表明,整个生长季代表真菌和G-的脂肪酸相对变化较明显。冗余分析(RDA,Redundancy analysis)表明,G+/G-比值与土壤温度呈正相关关系,土壤含水量与PLFA总量呈负相关关系,表明增温直接或间接导致G+/G-比值和PLFA总量变化,改变了土壤微生物的群落结构。     

长白山土壤微生物群落结构及酶活性随海拔的分布特征与影响因子

长白山是由火山喷发形成的山地生态系统,是研究生态系统重建和演替过程的天然实验室。以长白山西坡垂直带中的亚高山针叶林带、亚高山岳桦林带、高山草甸带、高山苔原带土壤为研究对象,采用磷脂脂肪酸法(PLFA)和微孔板法分别测定土壤微生物群落结构及酶活性,探讨(1)长白山西坡微生物群落结构及酶活性沿海拔的分布特征;(2)垂直带植被因子、土壤因子对微生物群落结构及酶活性的影响。结果表明:土壤有机碳、全氮、全磷含量均随海拔升高先增加再减少,有机碳和全氮最高值出现在岳桦林带;随海拔升高,土壤微生物总PLFA和各类群微生物PLFA呈现出先增加后减少的变化特征,表现为亚高山岳桦林带亚高山针叶林带高山草甸带高山苔原带;在对土壤微生物总PLFA的贡献率上,表现为细菌真菌放线菌,G~-G~+;微生物PLFA相关性方面,细菌、放线菌、G~+、G~-之间的关联性较大,真菌与这四者之间的关联性较小;土壤全氮含量与各微生物类群均表现为显著正相关,而C/N则与各微生物类群均表现为负相关,二者是调控土壤微生物沿海拔变化的主要因子;土壤水解酶βG和AP活性随海拔升高而逐渐增加,其中AP活性对高山苔原带生态系统表现出很好的响应;土壤含水量、C/N和土壤温度是调控土壤酶活性垂直变化的主要因子;高山苔原带草甸化过程对土壤含水量、全磷含量、水解酶AP活性产生重要影响,而对土壤微生物PLFA含量和其他酶活性影响不大。长白山垂直带土壤微生物群落结构和酶活性除了受到土壤环境因子和水热条件的影响,还与植被群落组成及凋落物性质具有紧密联系。     

Changes in Diversity and Functional Gene Abundances of Microbial Communities Involved in Nitrogen Fixation, Nitrification, and Denitrification in a Tidal Wetland versus Paddy Soils Cultivated for Different Time Periods

In many areas of China, tidal wetlands have been converted into agricultural land for rice cultivation. However, the consequences of land use changes for soil microbial communities are poorly understood. Therefore, we investigated bacterial and archaeal communities involved in inorganic nitrogen turnover (nitrogen fixation, nitrification, and denitrification) based on abundances and relative species richness of the corresponding functional genes along a soil chronosequence ranging between 50 and 2,000 years of paddy soil management compared to findings for a tidal wetland. Changes in abundance and diversity of the functional groups could be observed, reflecting the different chemical and physical properties of the soils, which changed in terms of soil development. The tidal wetland was characterized by a low microbial biomass and relatively high abundances of ammonia-oxidizing microbes. Conversion of the tidal wetlands into paddy soils was followed by a significant increase in microbial biomass. Fifty years of paddy management resulted in a higher abundance of nitrogen-fixing microbes than was found in the tidal wetland, whereas dominant genes of nitrification and denitrification in the paddy soils showed no differences. With ongoing rice cultivation, copy numbers of archaeal ammonia oxidizers did not change, while that of their bacterial counterparts declined. The nirK gene, coding for nitrite reductase, increased with rice cultivation time and dominated its functionally redundant counterpart, nirS, at all sites under investigation. Relative species richness showed significant differences between all soils with the exception of the archaeal ammonia oxidizers in the paddy soils cultivated for 100 and 300 years. In general, changes in diversity patterns were more pronounced than those in functional gene abundances.     

Accumulation of nitrogen and microbial residues during 2000 years of rice paddy and non‐paddy soil development in the Yangtze River Delta,China

Lowland rice paddy soils may accumulate significant amounts of organic matter. Our aim was to investigate the role of prolonged paddy management on the nitrogen (N) status of the soils, and to elucidate the contribution of bacteria and fungi to long‐term N accumulation processes. For this purpose, we sampled a chronosequence of 0–2000 years of rice cropping with adjacent non‐paddy systems in the Bay of Hangzhou, China. The samples were analyzed for bulk density, total, mineral and microbial N (N_mic), and amino sugars as markers for microbial residues. The results showed that during the first 100 years of land embankment, both paddy and non‐paddy soils accumulated N at a rate of up to 61 and 77 kg ha^?1 per annum, reaching steady‐state conditions after 110–172 years, respectively. Final N stocks in paddy fields exceeded those of the non‐paddies by a factor of 1.3. The contribution of amino sugars to total N increased to a maximum of 34 g N kg^?1 N in both land‐use systems, highlighting a significant accumulation of N in microbial residues of the surface soils. Correspondingly, the ratio of N_mic to microbial residue‐N decreased to a constant value. In the paddy subsoils, we found that bacterial residues particularly contributed to the pool of microbial residue‐N. Nevertheless, the absolute contents of amino sugars in paddy subsoils decreased during the last 1700 years of the chronosequence. We conclude that under paddy cultivation, soil microorganisms may accumulate parts of this N in their residues despite low overall N availability. However, this N accumulation is limited to initial stages of paddy soil development and restricted to the surface horizons, thus challenging its sustainability with future land‐use changes.     

石油污染土壤水改旱田后污染物组份及微生物群落结构变化

对沈抚灌区水改旱田不同年限土壤的石油污染物浓度及组分进行了分析,并采用变性梯度凝胶电泳和磷脂脂肪酸分析方法,分析了污染土壤微生物群落结构的变化.结果表明：1)石油污染土壤水改旱田后,年限越长,总多环芳烃在总石油烃中所占的比重越大,高分子量多环芳烃在总多环芳烃中所占比重也越大;2)总磷脂脂肪酸量与总石油烃呈显著正相关,与总多环芳烃相关性不显著;3)两种方法对土壤微生物群落结构的分析得出的结论一致,石油污染土壤微生物群落结构主要与其相对地理位置有关,当污染物的浓度达到一定程度时,土壤微生物群落结构会发生明显的改变.     

Molecular analysis of bacterial community structures in paddy soils for environmental risk assessment with two varieties of genetically modified rice, Iksan 483 and Milyang 204

The impacts of planted transgenic rice varieties on bacterial communities in paddy soils were monitored using both cultivation and molecular methods. The rice field plot consisted of eighteen subplots planted with two genetically modified (GM) rice and four non-GM rice plants in three replicates. Analysis with denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes revealed that the bacterial community structures were quite similar to each other in a given month, suggesting that there were no significant differences in bacterial communities between GM and non- GM rice soils. The bacterial community structures appeared to be generally stable with the seasons, as shown by a slight variation of microbial population levels and DGGE banding patterns over the year. Comparison analysis of 16S rDNA clone libraries constructed from soil bacterial DNA showed that there were no significant differences between GM and non-GM soil libraries but revealed seasonal differences of phyla distribution between August and December. The composition profile of phospholipid fatty acids (PLFA) between GM and non-GM soils also was not significantly different to each other. When soil DNAs were analyzed with PCR by using primers for the bar gene, which was introduced into GM rice, positive DNA bands were found in October and December soils. However, no bar gene sequence was detected in PCR analysis with DNAs extracted from both cultured and uncultured soil bacterial fractions. The result of this study suggested that, in spite of seasonal variations of bacterial communities and persistence of the bar gene, the bacterial communities of the experimental rice field were not significantly affected by cultivation of GM rice varieties.     

不同稻作制对红壤性水稻土中锰剖面分布的影响

为了探讨红壤性水稻土锰的迁移和转化行为,通过长达16a的定位试验研究了不同稻作制、有机肥以及地下水位对土壤剖面中全锰、活性锰和交换态锰分布的影响。试验结果表明,长期淹水种稻引起0-20cm土壤层次全锰、活性锰和交换态锰含量的显著下降,而在20－40cm和40－75cm土层相对累积。不同稻作制比较,0－20cm土层中全锰、活性锰和交换态锰含量以稻稻泡显著高于稻稻绿和稻稻油处理,而20－40cm和40－75cm土层3种锰形态的含量各稻作制之间无显著差异,表明实行水旱轮作的稻稻绿与稻稻油两种稻作制耕层土壤锰的淋溶损失比持续淹水的稻稻泡制更为严重。相对而言,不同有机肥施用水平和地下水位对土壤剖面中锰分布的影响要小于稻作制,总的趋势为：土壤剖面中锰的空间分异程度以高量有机肥＞常量有机肥＞单施化肥;低水位＞高水位。从土壤中锰的空间分布规律可以看出,水旱轮作（尤其是在低水位和有机无机配合的条件下）比长期淹水更有利于土壤锰氧化还原引起的深层淋溶与淀积,加速了典型水稻土剖面的形成。     

冬季覆盖作物秸秆还田对双季稻田根际土壤微生物群落功能多样性的影响

微生物群落功能多样性是土壤质量变化重要的指标,不同作物类型的秸秆还田措施对土壤微生物群落功能多样性具有明显的影响。以位于双季稻主产区不同冬季覆盖作物-双季稻种植模式大田定位试验田为研究对象,以冬闲-双季稻种植模式为对照(CK),应用Biolog-GN技术开展黑麦草-双季稻(Ry)、紫云英-双季稻(Mv)、油菜-双季稻(Ra)和马铃薯-双季稻(Po)种植模式条件下不同冬季覆盖作物秸秆还田后对双季稻田根际土壤微生物功能多样性影响的研究。研究结果表明,早稻和晚稻成熟期,与CK处理相比,冬季覆盖作物秸秆还田处理增加了稻田土壤碳源平均颜色变化率(AWCD),以Po处理AWCD均为最高,均显著高于Ry和CK处理。不同冬季覆盖作物秸秆还田处理土壤微生物代谢多样性指数表现出明显的差异,早稻成熟期,Po处理的Richness、Shannon和McIntosh指数均为最高,其次为Ry、Mv和Ra处理,CK处理最低;晚稻成熟期,各处理的Richness、Shannon和McIntosh指数大小顺序均表现为PoRaMvRyCK。土壤微生物碳源利用的主成分分析结果表明,各冬季覆盖作物秸秆还田处理根际土壤微生物利用的主要碳源为氨基酸类和糖类物质,不同处理间碳源利用类型有差异。冬季覆盖作物秸秆还田措施有利于提高双季稻田根际土壤微生物对碳源的利用能力、物种丰富度和均匀度。     

Assessing the effect of air-drying and storage on microbial biomass and community structure in paddy soils

To understand the effect of air-drying pre-treatment, refrigeration, and freezing storages on microbial biomass and community structure in paddy soils, we measured total phospholipid fatty acid (PLFA) and PLFA profile after five treatments, including flooded (F), flooded-freezing (FF), flooded-air-drying (FAD), flooded-air-drying-freezing (FADF), and flooded-air-drying-refrigeration (FADR). FF and FADF treatments were followed by freeze-drying before analyzing the total PLFA and PLFA profile. The results showed that FF and FADF treatments increased the content of polyunsaturated fatty acids, but decreased that of branched chain saturated fatty acids. FAD treatment increased the concentrations of bacterial, aerobic bacterial, stress, Type I methanotrophs, and Gram-negative bacterial biomarkers, while it decreased the concentration of hydroxy fatty acid group and the ratios of cyclopropyl saturated fatty acids to their monoenoic precursors. FADR significantly decreased the concentration of total PLFA and all PLFA groups except for the mono-unsaturated fatty acid group. Statistical analysis with correspondence analysis showed that air-drying and storage changed the microbial community structure, but the effect of air-drying on soil microbial community structure was more pronounced than that of freezing. These results indicated that deep freezing followed by freeze-drying may be the most recommendable procedure before soil biochemical analysis in flooded paddy soils.     

Characterization of Microbial Consortia in Paddy Rice Soil by Phospholipid Analysis

Abstract Microbial biomass and community structure in paddy rice soil during the vegetation period of rice were estimated by analysis of their phospholipid fatty acids (PLFA), hydroxy fatty acids of lipopolysaccharides (LPS-HYFA), and phospholipid ether lipids (PLEL) directly extracted from the soil. A clear change in the composition of the community structure at different sampling periods was observed, indicated by the principal component analysis of the PLFA. A dramatic decline of ester-linked PLFA was observed in the soil samples taken at the second sampling time. In contrast to the ester-linked PLFA, the non-ester-linked PLFA composition did not change. The hydroxy fatty acids of lipopolysaccharides as well as ether lipids decreased consecutively during the observation period. Total microbial abundance was estimated to be (4.1–7.3) × 10⁹ cells g^-1 soil (dry weight). About 44% account for aerobic and 32% for facultative anaerobic bacteria, and 24% for archaea, on average. According to the profile and patterns of PLFA in the soil sample, it may be suggested that the paddy soil at the August sampling period contained more abundant facultative anaerobic bacteria (ca. 36%) and archaea (ca. 37%), but the total microbial biomass was significantly lower than in the remaining sampling periods. As the plant approached maturity, the microbial community structure in the soil changed to contain more abundant Gram-negative bacteria and methanotrophs. Received: 23 September 1999; Accepted: 28 February 2000; Online Publication: 12 May 2000     

Bacterial succession during 500?years of soil development under agricultural use

Soil bacterial succession under intensive anthropogenic disturbances is not well known. Using terminal restriction fragment length polymorphisms and 454 pyrosequencing of 16S rRNA genes, this study investigated how soil bacterial diversity and community structure changed under two agricultural land uses (paddy rice and upland cropping) in relation to soil development along a 500-year chronosequence created by intermittent reclamation of estuarine salt marshes. Multivariate analysis revealed orderly changes in soil physicochemical properties and bacterial community structure with time, confirming the occurrence of soil development and bacterial succession. Patterns of soil development and bacterial succession resembled each other, with recent land uses affecting their trajectories but not the overall direction. Succession of bacterial community structure was mainly associated with changes in ??-Proteobacteria and Verrucomicrobia. Two stages of bacterial succession were observed, a dramatic-succession stage during the first several decades when bacterial diversity increased evidently and bacterial community structure changed rapidly, and a long gradual-succession stage that lasted for centuries. Canonical correspondence analysis identified soil Na⁺, potentially mineralizable nitrogen, total phosphorous, and crystallinity of iron oxyhydrates as potential environmental drivers of bacterial succession. To conclude, orderly succession of soil bacterial communities occurred along with the long-term development of agroecosystems, which in turn was associated with soil physicochemical changes over time.     

Rice to Vegetables: Short- Versus Long-Term Impact of Land-Use Change on the Indigenous Soil Microbial Community

Land-use change is known to have a significant effect on the indigenous soil microbial community, but it is unknown if there are any general trends regarding how this effect varies over time. Here, we describe a comparative analysis of microbial communities from three adjacent agricultural fields: one-century-old paddy field (OP) and two vegetable fields (new vegetable field (NV) and old vegetable field (OV)) that were established on traditional paddy fields 10 and 100 years ago, respectively. Soil chemical and physical analysis showed that both vegetable fields were more nutrient rich than the paddy field in terms of organic C, total N, total P, and available K. The vegetable fields possessed relatively higher abundance of culturable bacteria, fungi, and specific groups of bacteria (Actinomyces, nitrifying bacteria, and cellulose-decomposing bacteria) but lower levels of microbial biomass C and N. Notably, the decrease of biomass was further confirmed by analysis of seven additional soils in chronosequence sampled from the same area. Next we examined the metabolic diversity of the microbial community using the EcoPlate^TM system from Biolog Inc. (Hayward, CA, USA). The utilization patterns of 31 unique C substrates (i.e., community-level physiological profile) showed that microorganisms in vegetable soil and paddy soil prefer to use different C substrates (polymeric compounds for NV and OV soils, phenolic acids for OP soil). Principal component analysis and the average well color development data showed that the NV is metabolically more distinct from the OV and OP. The effect was likely attributable to the elevated soil pH in NV soil. Furthermore, we assessed the diversity of soil bacterial populations using the cultivation-independent technology of amplified ribosomal DNA restriction analysis (ARDRA). Results showed that levels of bacterial diversity in OP and NV soils were similar (Shannon’s diversity index H = 4.83 and 4.79, respectively), whereas bacteria in OV soil have the lowest score of diversity (H = 3.48). The low level of bacterial diversity in OV soil was supported by sequencing of ten randomly selected 16S rDNA clones from each of the three rDNA libraries. Phylogenetic analysis showed that all the ten OV clones belonged to Proteobacteria with eight in the gamma-subdivision and two in the alpha-subdivision. In contrast, the ten clones from NV and OP soils were classified into four and eight bacterial classes or unclassified groups, respectively. Taken together, our data suggest that land-use change from rice to vegetables resulted in a decrease of bacterial diversity and soil biomass despite an increase in the abundance of culturable microorganisms and, moreover, the decrease of bacterial diversity occurred during long-term rather than short-term vegetable cultivation.