CO2浓度增加对长白山森林土壤甲烷氧化影响

大气CO2浓度升高可能对森林土壤的甲烷(CH4)氧化速率产生影响.本文采用开顶箱技术,对连续6年高浓度CO2(500 μmol·mol-1)处理的长白山森林典型树种蒙古栎树下土壤CH4氧化速率进行研究,并利用CH4氧化菌的16S rRNA特异性引物以及CH4单加氧酶功能基因引物分析了土壤中CH4氧化菌的群落结构与数量.结果表明:CO2浓度增高后,生长季土壤甲烷氧化量与对照和裸地相比分别降低了4％和22％;基于16S rRNA特异性引物的DGGE分析表明,CO2浓度增高导致两类甲烷氧化菌的多样性指数降低;CO2浓度增高对土壤中Ⅰ类甲烷氧化菌数量无显著影响,而使土壤中Ⅱ类甲烷氧化菌数量显著减少,功能基因pmoA拷贝数与对照和裸地相比分别降低了15％和46％.CO2浓度增高导致森林土壤甲烷氧化菌数量与活性降低,土壤含水量的增加可能是导致这一现象的主要原因.     

CO2浓度增加对长白山森林土壤甲烷氧化影响

大气CO2浓度升高可能对森林土壤的甲烷(CH4)氧化速率产生影响.本文采用开顶箱技术,对连续6年高浓度CO2（500 μmol·mol-1）处理的长白山森林典型树种蒙古栎树下土壤CH4氧化速率进行研究,并利用CH4氧化菌的16S rRNA特异性引物以及CH4单加氧酶功能基因引物分析了土壤中CH4氧化菌的群落结构与数量.结果表明： CO2浓度增高后,生长季土壤甲烷氧化量与对照和裸地相比分别降低了4%和22%;基于16S rRNA特异性引物的DGGE分析表明,CO2浓度增高导致两类甲烷氧化菌的多样性指数降低;CO2浓度增高对土壤中Ⅰ类甲烷氧化菌数量无显著影响,而使土壤中Ⅱ类甲烷氧化菌数量显著减少,功能基因pmoA拷贝数与对照和裸地相比分别降低了15%和46%.CO2浓度增高导致森林土壤甲烷氧化菌数量与活性降低,土壤含水量的增加可能是导致这一现象的主要原因.     

遮阴对米槠和杉木原位排放甲烷的影响

尽管植物在有氧环境是否有氧排放甲烷(CH4)一直存在很大的争议,但越来越多的研究证实植物自身可以排放CH4,而光照是影响植物排放CH4的重要因素。利用13C同位素标记技术结合盆栽实验,探讨了米槠和杉木叶片原位排放CH4的来源及其对太阳光照和遮阴处理的响应。结果表明,标记米槠和杉木叶片的碳稳定同位素13C值分别比未标记的高128.9和71.1倍;密封标记米槠和杉木叶片2h后的δ13CH4值分别比0h时高21.1倍和28.2倍,而未标记米槠和杉木在密封0h与2h时的δ13CH4值之间均没有显著差异,这证实了米槠和杉木排放的CH4主要源于他们自身。自然光照下,杉木的平均CH4排放速率比米槠的平均CH4排放速率高29%。自然光照下的米槠和杉木的CH4排放速率具有相同的波动规律;遮阴对米槠和杉木的CH4排放的影响不同,遮阴处理米槠的CH4排放速率显著低于自然光照处理,平均低23%;遮阴处理杉木的平均CH4排放速率与自然光照处理的没有显著差异,但在遮阴处理5d之后,其CH4排放速率比自然光照下的高71%,之后其CH4排放速率急剧下降,且明显低于自然光照下的CH4排放速率。以上结果表明,太阳光照促进了植物排放CH4的速率。     

长期不同施肥下水稻土甲烷氧化能力及甲烷氧化菌多样性的变化

稻田内源甲烷的氧化是稻田甲烷减排的重要途径。而甲烷氧化菌是土壤中甲烷氧化的主要施动者,在长期不同施肥条件下,土壤微生物群落的演变是否影响到土壤甲烷氧化菌群落结构及其活性,进而影响到田土壤CH4向大气的实际排放强度还不清楚。为此,选择太湖地区一个长期肥料试验的稻田土壤为研究对象,分析长期不同肥料施用对土壤甲烷氧化能力的影响及其与土壤中甲烷氧化菌群落结构变化的可能关系。结果表明,长期不同的施肥措施下稻田土壤对甲烷的氧化能力产生了明显差异,伴随着土壤中甲烷氧化菌（MOBI和MOBII）的基因群落多样性的显著变化。长期单一施用氮肥为主的化肥显著降低了土壤对甲烷的氧化能力,同时显著降低了稻田土壤甲烷氧化菌的多样性和丰富度;不同施肥下甲烷氧化菌多样性的变化与土壤的甲烷氧化能力的变化趋势相一致。因此,研究显示长期不同施肥处理下甲烷氧化菌群落结构的改变可能是引起水稻土甲烷氧化能力变化的一个主要因素,有机无机配合施用可以明显降低稻田土壤甲烷的大气释放潜能。但长期不同施肥处理下甲烷氧化菌活性的变化还有待于进一步研究。     

氮沉降对温带森林土壤甲烷氧化菌的影响

大量研究显示氮沉降影响森林甲烷吸收量,但其中的微生物驱动机制仍缺乏研究。基于长白山典型温带森林长期氮沉降模拟实验平台样地,采用定量PCR和克隆测序技术,研究了长期施加不同形态氮((NH_4)_2SO_4、NH_4Cl和KNO_3)处理下森林土壤甲烷氧化菌的数量和群落组成随季节变化的特征。结果表明,夏季,森林土壤甲烷氧化菌pmo A基因丰度在不同施氮处理之间无显著性差异(每克干土1.54×10~6-3.20×10~6拷贝数);秋季,pmo A基因丰度在施加NH_4Cl和(NH_4)_2SO_4处理小区(每克干土1.93×10~5-7.6×10~5拷贝数)与对照(每克干土(4.03×10~6±1.2×10~6)拷贝数)相比有所降低,尤其在(NH_4)_2SO_4处理小区(每克干土(4.61×10~5±2.61×10~5)拷贝数)显著降低;无论夏季还是秋季,施加不同形态氮处理土壤甲烷氧化菌均以Type I型为主(相对丰度在70.6%-85.4%之间),并以Methylobacter-group(Type I)为优势类群,占Type I型的55.1%-91.7%;Methylobacter-group(Type I)的相对丰度在夏季不同形态氮处理土壤样品中无显著差异,但秋季样品中在施加(NH_4)_2SO_4(52.7%±6.5%)和NH_4Cl(56.1%±8.9%)的处理显著低于对照土壤(77.0%±2.9%),Methylococcus-group(Type I)的相对丰度则在(NH_4)_2SO_4和NH_4Cl处理土壤呈增加的趋势。这些结果表明铵态氮肥添加对温带森林土壤甲烷氧化菌的生长具有抑制作用并导致其群落结构发生改变,受夏季温度和水分的影响,这种抑制作用在秋季表现更明显,而NO_3~--N添加对土壤甲烷氧化菌的群落组成和丰度无显著影响。这些结果解释了以往观测到的施铵态氮肥显著降低秋季温带林地土壤甲烷净吸收量,而在夏季无显著影响的观测结果,解释了长期氮沉降影响森林土壤甲烷吸收的微生物机制。     

干旱胁迫对不同根型苜蓿根系生长及根际土壤细菌的影响

为明确干旱胁迫对根茎型清水紫花苜蓿、直根型陇东紫花苜蓿、根蘖型公农4号杂花苜蓿根系生长及根际土壤细菌群落的影响,采用盆栽试验,运用16S rRNA基因测序技术,研究了幼苗期干旱胁迫下各根型苜蓿根系生长及根际土壤细菌群落结构的变化。结果表明：干旱胁迫下各根型苜蓿的Chao1和ACE丰富度指数均在中度胁迫下最大,Simpson和Shannon-wiener多样性指数各处理间差异不显著;根际土壤细菌群落均以变形菌门、绿弯菌门、类杆菌门、放线菌门和厚壁菌门为主,干旱胁迫均显著增加了变形菌门和厚壁菌门的相对丰度,显著降低了绿弯菌门的相对丰度,但类杆菌门和放线菌门先提高后下降。统计学分析显示,幼苗期干旱胁迫显著影响各根型苜蓿生长发育,随胁迫程度增加,其株高、地上生物量、地下生物量、根系活力、根体积、根系总长均显著降低,根冠比先增加后下降且在中度胁迫时达到最大值。重度胁迫下,清水苜蓿的株高、根系活力显著大于其他品种,而根冠比、根系干重显著小于其他品种;陇东苜蓿的根长、根尖数均显著大于其他品种;根系平均直径、根系总表面积各根型苜蓿间差异不显著。研究结果为植物的抗干旱胁迫以及提高各根型苜蓿在干旱胁迫下的水分利用提供参考。     

温度对亚热带地区常见树种叶片甲烷排放的影响

以亚热带常见树种米槠、木荷、浙江桂、罗浮栲、杉木和柑橘为对象,利用控制试验研究了温度对树木叶片甲烷(CH4)排放的影响.结果表明:当温度在10℃时,供试的6种树木中,仅木荷、柑橘和罗浮栲的叶片排放CH4;温度高于20℃时,所有树木叶片均可排放CH4.温度高于30℃时,叶片排放CH4的平均排放速率(1.010ngCH4·g-1DM·h-1)是10～30℃时平均排放速率(0.255ngCH4·g-1DM·h-1)的3.96倍.增温对柑橘和杉木CH4排放速率的影响显著高于其他4种树木.培养时间对叶片排放CH4速率有显著影响,温度胁迫对树木排放CH4的影响受植物活性的控制.在低温或高温条件下,树木干叶均不能排放CH4.高温胁迫对树木叶片排放CH4有重要影响,全球变暖可能增加植物的CH4排放.     

不饱和土壤CH4的吸收与氧化

不饱和土壤是已知唯一的 CH4 生物壑。综述了不饱和土壤 CH4 的吸收、氧化过程及其影响因素。不饱和土壤中 CH4 氧化的临界浓度低 ,因而甲烷氧化菌可氧化大气 CH4 并将其当作唯一的碳源和能源。土壤 CH4 吸收率与土壤湿度通常呈负相关关系。土壤湿度过高 ,大气 CH4 和 O2 向土壤中扩散受阻 ;或土壤湿度过低引起水分胁迫均导致甲烷氧化菌活性下降。NH 4对土壤中 CH4 氧化的抑制作用可归结为 NH3和 CH4 在甲烷单氧酶水平上的竞争、由氧化作用向硝化作用的转移以及 NH 4氧化生成的 NO- 2 的毒性。NH 4对 CH4 氧化的抑制作用与土壤有效氮含量成正比。各类氮肥对 CH4 氧化抑制作用 :化肥 >有机肥 ;铵态氮肥 >尿素。 NO- 3对 CH4 氧化没有抑制效应。阳离子代换量 (CEC)高的土壤 NH 4对 CH4 氧化的抑制作用轻。 CH4 氧化菌对大气 CH4 的高亲和力及 CH4 氧化所需较低的活化能导致其温度系数 Q1 0 较小。地温较低时 ,土壤氧化 CH4 的能力随温度升高而升高。当地温高于 CH4 氧化的最佳温度时 ,CH4 氧化菌难以与硝化细菌及其它微生物竞争利用土壤空气中的 O2 ,导致其活性降低。甲烷氧化菌对 p H值变化不敏感。团粒结构较好的壤土可保护 CH4 氧化菌免受干扰。未受干扰的森林土壤 CH4 氧化率的峰值一般出现在亚表     

黄土高原半干旱区不同种植年限紫花苜蓿人工草地土壤微生物和线虫群落特征

建植紫花苜蓿人工草地是黄土高原植被恢复的重要措施之一。土壤微生物和线虫群落特征是评价和调控植被恢复的生态环境效应的重要依据。本研究在宁夏南部山区选取不同种植年限(1、2、6和12年)的紫花苜蓿人工草地为研究样地,以农田和天然草地作为对照,探索黄土高原人工草地植被恢复过程中土壤微生物和线虫群落的演变规律及其影响因素。结果表明: 1)种植苜蓿显著提高了土壤细菌群落的Chao1、ACE和Shannon多样性指数,并在种植苜蓿后第6年达到最高,但在种植6年和12年后真菌群落多样性降低;随着苜蓿种植年限的增加,真菌群落组成从农田逐渐向天然草地方向演变;2)土壤线虫数量与细菌群落多样性的变化趋势相同,在种植苜蓿后第6年出现峰值,该时期线虫群落结构组成与农田较相似,苜蓿12年样地则更接近天然草地;随着苜蓿种植年限的增加,食细菌线虫、植食性线虫比例总体呈上升趋势,食真菌线虫、杂食/捕食线虫比例呈下降趋势,土壤成熟度指数(MI)逐渐减小,植物寄生线虫指数(PPI)和线虫通路指数(NCR)则不断增大;3)在苜蓿人工草地植被恢复过程中,土壤有机碳、全氮和速效磷对土壤微生物群落结构影响较大,并进一步影响线虫群落结构;细菌和真菌群落优势类群和多样性与线虫的不同营养类群及生态指数之间存在密切联系,表明微生物群落结构与多样性对线虫群落具有显著影响;在不同种植年限苜蓿草地中,植物的生物量与多样性的变化可能通过影响土壤微生物与线虫食物资源状况从而引起其群落特征的改变。     

三江源地区不同建植期人工草地植被特征及其与土壤特征的关系

研究了三江源地区不同建植期人工草地群落生物量、物种组成、多样性指数和土壤理化特征,并用多元逐步回归分析法探讨了土壤理化特征对群落生物量、多样性变化的响应.结果表明:研究区不同建植期人工草地植物群落的种类组成、植物功能群组成和群落数量特征存在显著差异;土壤含水量随着物种多样性指数的增加而增加,土壤容重随着物种多样性的增加而减小;土壤微生物生物量碳与土壤含水量、土壤有机质呈极显著正相关,与土壤容重呈极显著负相关;土壤有机碳含量明显呈"V"字型变化,且与土壤含水量的变化趋势相一致,随土壤容重的增加而减少;群落生物量与土壤养分和土壤含水量之间呈显著正相关,群落地上、地下生物量的增加有利于提高土壤养分含量.     

Optimization of diagnostic microarray for application in analysing landfill methanotroph communities under different plant covers

Landfill sites are responsible for 6-12% of global methane emission. Methanotrophs play a very important role in decreasing landfill site methane emissions. We investigated the methane oxidation capacity and methanotroph diversity in lysimeters simulating landfill sites with different plant vegetations. Methane oxidation rates were 35 g methane m-2 day-1 or higher for planted lysimeters and 18 g methane m-2 day-1 or less for bare soil controls. Best methane oxidation, as displayed by gas depth profiles, was found under a vegetation of grass and alfalfa. Methanotroph communities were analysed at high throughput and resolution using a microbial diagnostic microarray targeting the particulate methane monooxygenase (pmoA) gene of methanotrophs and functionally related bacteria. Members of the genera Methylocystis and Methylocaldum were found to be the dominant members in landfill site simulating lysimeters. Soil bacterial communities in biogas free control lysimeters, which were less abundant in methanotrophs, were dominated by Methylocaldum. Type Ia methanotrophs were found only in the top layers of bare soil lysimeters with relatively high oxygen and low methane concentrations. A competetive advantage of type II methanotrophs over type Ia methanotrophs was indicated under all plant covers investigated. Analysis of average and individual results from parallel samples was used to identify general trends and variations in methanotroph community structures in relation to depth, methane supply and plant cover. The applicability of the technology for the detection of environmental perturbations was proven by an erroneous result, where an unexpected community composition detected with the microarray indicated a potential gas leakage in the lysimeter being investigated.     

Comparison of RNA- and DNA-based bacterial communities in a lab-scale methane-degrading biocover

Methanotrophs must become established and active in a landfill biocover for successful methane oxidation. A lab-scale biocover with a soil mixture was operated for removal of methane and nonmethane volatile organic compounds, such as dimethyl sulfide (DMS), benzene (B), and toluene (T). The methane elimination capacity was 211?±?40 g?m^?2 d^?1 at inlet loads of 330–516 g?m^?2 d^?1. DMS, B, and T were completely removed at the bottom layer (40–50 cm) with inlet loads of 221.6?±?92.2, 99.6?±?19.5, and 23.4?±?4.9 mg m^?2 d^?1, respectively. The bacterial community was examined based on DNA and RNA using ribosomal tag pyrosequencing. Interestingly, methanotrophs comprised 80 % of the active community (RNA) while 29 % of the counterpart (DNA). Types I and II methanotrophs equally contributed to methane oxidation, and Methylobacter, Methylocaldum, and Methylocystis were dominant in both communities. The DNA vs. RNA comparison suggests that DNA-based analysis alone can lead to a significant underestimation of active members.     

Identity of active methanotrophs in landfill cover soil as revealed by DNA-stable isotope probing

A considerable amount of methane produced during decomposition of landfill waste can be oxidized in landfill cover soil by methane-oxidizing bacteria (methanotrophs) thus reducing greenhouse gas emissions to the atmosphere. The identity of active methanotrophs in Roscommon landfill cover soil, a slightly acidic peat soil, was assessed by DNA-stable isotope probing (SIP). Landfill cover soil slurries were incubated with (13)C-labelled methane and under either nutrient-rich nitrate mineral salt medium or water. The identity of active methanotrophs was revealed by analysis of (13)C-labelled DNA fractions. The diversity of functional genes (pmoA and mmoX) and 16S rRNA genes was analyzed using clone libraries, microarrays and denaturing gradient gel electrophoresis. 16S rRNA gene analysis revealed that the cover soil was mainly dominated by Type II methanotrophs closely related to the genera Methylocella and Methylocapsa and to Methylocystis species. These results were supported by analysis of mmoX genes in (13)C-DNA. Analysis of pmoA gene diversity indicated that a significant proportion of active bacteria were also closely related to the Type I methanotrophs, Methylobacter and Methylomonas species. Environmental conditions in the slightly acidic peat soil from Roscommon landfill cover allow establishment of both Type I and Type II methanotrophs.     

Activity of a Methanotrophic Consortium Isolated from Landfill Cover Soil: Response to Temperature,pH, CO2, and Porous Adsorbent

A robust, naturally evolving methanotrophic community in landfill cover soil (LFCS) can be the simplest way to mitigate landfill methane emission. In this study, bacterial community composition in LFCS and methane oxidation potential of enriched methanotrophic consortium, in comparison to that of axenic Methylosinus sporium, was investigated. Growth and methane oxidation of the consortium was studied in liquid phase batch experiments under varying temperature (20–40°C), pH (5–10), headspace CO2, and in presence of porous adsorbent (1.3 cm³ sponge cubes). The 16S rRNA gene analysis revealed presence of both type-I and type-II methanotrophs along with few obligate methylotroph in LFCS. Though the optimal growth condition of the consortium was at 30°C and pH 7, it was more resilient in comparison to M. sporium. With increasing availability of porous adsorbent, methane consumption by the consortium was significantly improved (p < 0.001) reaching a maximum specific methane oxidation rate of 11.4 μmol mg^?1 biomass h^?1. Thus, inducing naturally thriving methanotrophs in LFCS is a better alternative to axenic methanotrophic culture in methane emission management.     

Effect of afforestation and reforestation of pastures on the activity and population dynamics of methanotrophic bacteria

We investigated the effect of afforestation and reforestation of pastures on methane oxidation and the methanotrophic communities in soils from three different New Zealand sites. Methane oxidation was measured in soils from two pine (Pinus radiata) forests and one shrubland (mainly Kunzea ericoides var. ericoides) and three adjacent permanent pastures. The methane oxidation rate was consistently higher in the pine forest or shrubland soils than in the adjacent pasture soils. A combination of phospholipid fatty acid (PLFA) and stable isotope probing (SIP) analyses of these soils revealed that different methanotrophic communities were active in soils under the different vegetations. The C18 PLFAs (signature of type II methanotrophs) predominated under pine and shrublands, and C16 PLFAs (type I methanotrophs) predominated under pastures. Analysis of the methanotrophs by molecular methods revealed further differences in methanotrophic community structure under the different vegetation types. Cloning and sequencing and terminal-restriction fragment length polymorphism analysis of the particulate methane oxygenase gene (pmoA) from different samples confirmed the PLFA-SIP results that methanotrophic bacteria related to type II methanotrophs were dominant in pine forest and shrubland, and type I methanotrophs (related to Methylococcus capsulatus) were dominant in all pasture soils. We report that afforestation and reforestation of pastures caused changes in methane oxidation by altering the community structure of methanotrophic bacteria in these soils.     

Effect of Afforestation and Reforestation of Pastures on the Activity and Population Dynamics of Methanotrophic Bacteria

We investigated the effect of afforestation and reforestation of pastures on methane oxidation and the methanotrophic communities in soils from three different New Zealand sites. Methane oxidation was measured in soils from two pine (Pinus radiata) forests and one shrubland (mainly Kunzea ericoides var. ericoides) and three adjacent permanent pastures. The methane oxidation rate was consistently higher in the pine forest or shrubland soils than in the adjacent pasture soils. A combination of phospholipid fatty acid (PLFA) and stable isotope probing (SIP) analyses of these soils revealed that different methanotrophic communities were active in soils under the different vegetations. The C₁₈ PLFAs (signature of type II methanotrophs) predominated under pine and shrublands, and C₁₆ PLFAs (type I methanotrophs) predominated under pastures. Analysis of the methanotrophs by molecular methods revealed further differences in methanotrophic community structure under the different vegetation types. Cloning and sequencing and terminal-restriction fragment length polymorphism analysis of the particulate methane oxygenase gene (pmoA) from different samples confirmed the PLFA-SIP results that methanotrophic bacteria related to type II methanotrophs were dominant in pine forest and shrubland, and type I methanotrophs (related to Methylococcus capsulatus) were dominant in all pasture soils. We report that afforestation and reforestation of pastures caused changes in methane oxidation by altering the community structure of methanotrophic bacteria in these soils.     

Abundance, distribution and potential activity of methane oxidizing bacteria in permafrost soils from the Lena Delta, Siberia

The methane oxidation potential of active layer profiles of permafrost soils from the Lena Delta, Siberia, was studied with regard to its respond to temperature, and abundance and distribution of type I and type II methanotrophs. Our results indicate vertical shifts within the optimal methane oxidation temperature and within the distribution of type I and type II methanotrophs. In the upper active layer, maximum methane oxidation potentials were detected at 21 degrees C. Deep active layer zones that are constantly exposed to temperatures below 2 degrees C showed a maximum potential to oxidize methane at 4 degrees C. Our results indicate a dominance of psychrophilic methanotrophs close to the permafrost table. Type I methanotrophs dominated throughout the active layer profiles but their number strongly fluctuated with depth. In contrast, type II methanotrophs were constantly abundant through the whole active layer and displaced type I methanotrophs close to the permafrost table. No correlation between in situ temperatures and the distribution of type I and type II methanotrophs was found. However, the distribution of type I and type II methanotrophs correlated significantly with in situ methane concentrations. Beside vertical fluctuations, the abundance of methane oxidizers also fluctuated according to different geomorphic units. Similar methanotroph cell counts were detected in samples of a flood plain and a polygon rim, whereas cell counts in samples of a polygon centre were up to 100 times lower.     

Community structure in a methanotroph biofilter as revealed by phospholipid fatty acid analysis

The microbial community structure of two biofilters used for the oxidation of methane and organic trace gases generated in landfills was analysed by phospholipid fatty acid composition. Community structure varied with biofilter depth, reflecting varying conditions of substrate supply as well as of organic carbon content, nutrient status and osmotic stress determined by the different materials used for the individual biofilter layers. Both biofilters were dominated by type II methanotrophs. In the biofilter charged with landfill gas containing significant amounts of trace organics, fatty acid 18:1omega7c constituted 87% of the methanotrophic PLFA, while the recognised signature fatty acids 16:1omega8 and 18:1omega8, which were well represented in the other biofilter, were entirely absent. This indicates the development of a highly specific methanotrophic population, presumably as a result of the adaption to continuous organic trace gas exposure.