藏北高原草甸土壤固碳微生物群落特征随海拔和季节的变化

研究土壤固碳微生物丰度、群落结构、多样性差异及其影响因子对了解青藏高原土壤碳循环和固碳潜力具有重要意义。采用定量PCR(qPCR)、末端限制性片段分析(T-RFLP)、克隆文库和测序方法,研究了青藏高原草甸土壤固碳微生物丰度与群落结构随海拔和季节的变化,主要结果如下:1)随海拔升高高寒草甸土壤固碳微生物丰度显著升高,但季节变化不明显,不同类别微生物固碳基因cbbL丰度依次为:Form ICForm IABForm ID,其中Form IC类固碳微生物可达10~8拷贝数/g土壤,cbbL基因丰度与海拔、土壤含水量和铵态氮含量(NH_4~+-N)呈正相关关系,与土壤温度和pH值负相关;2)固碳微生物多样性和丰富度随海拔升高而升高,在4800m达到最大,且二者受季节影响较小,其群落结构随海拔升高而逐渐变化,主要受土壤pH值、海拔和土壤水分影响;3)Form IC类固碳微生物主要包括放线菌门和和变形菌门,其中α变形菌门是高寒草甸土壤优势固碳微生物类群。本研究有助于理解土壤微生物群落功能及其在土壤碳循环过程中的作用,为更准确评估高寒草甸土壤碳循环过程提供了科学依据。     

毛竹林集约经营对土壤固碳细菌群落结构和多样性的影响

为揭示毛竹集约经营对土壤固碳细菌的影响,分别采集集约经营时间为0、10、15、20年和25年的毛竹林土壤(0—20 cm和20—40 cm)土壤,应用实时荧光定量PCR、T-RFLP以及cbbL基因文库方法,分析毛竹林长期集约经营过程中土壤固碳细菌丰度和群落结构多样性的变化,通过冗余分析(RDA)探讨影响土壤固碳细菌群落的主要环境因素。结果表明,长期的集约经营显著提高了毛竹林表层和亚表层土壤的养分含量,土壤pH值却明显降低。集约经营毛竹林土壤固碳微生物数量并未表现出与SOC的相关性,而与N素水平的变化显著相关。具体表现为:随着集约经营的进行表层cbbL基因丰度呈先上升(10年)后下降的规律,与氮素水平呈正相关(P0.05);亚表层土壤cbbL基因丰度则呈直线下降的趋势,与C∶N呈正相关(P0.05)。集约经营导致表层和亚表层土壤微生物群落结构改变,表层固碳细菌多样性指数下降。由系统发育分析可知,不可培养固碳细菌占56%比例,土壤中共同的优势种类多为变形菌和放线菌,以兼性自养为主。RDA分析结果表明土壤酸化和养分积累是毛竹林土壤固碳细菌群落和多样性变化的重要原因。     

水分含量对紫色土自养微生物丰度与活性的影响

通过室内培养试验,研究土壤水分对紫色土自养微生物丰度与活性的影响。土壤含水量(质量百分比)设置为4个处理,分别为10%(W_(10))、15%(W_(15))、25%(W_(25))及淹水(WL),于室温下培养28天。分别于培养的第3、7、14、28天取样,测定土壤有机碳含量、cbb L基因(1,5-二磷酸核酮糖羧化酶大亚基基因)丰度、RubisCO(1,5-二磷酸核酮糖羧化酶)酶活性,并分析它们之间的相关性。结果表明:土壤cbbL基因丰度于培养的前14天呈现随含水量增加而升高的趋势,且淹水处理(WL)在整个培养期间其cbbL丰度均处于最高水平。各处理土壤的RubisCO酶活性在培养初期(第3天)无显著差异,培养后期(第28天)低含水量(W_(10)和W_(15))处理中RubisCO酶活性显著低于高含水量的W_(25)及WL处理(P0.05)。单因素重复检测方差分析结果表明,含水量显著影响土壤有机碳含量(F=11.91,P=0.03)、cbbL丰度(F=42.33,P0.01)和RubisCO酶活性(F=51.55,P0.01)。相关性分析表明,土壤cbbL丰度与RubisCO酶活性呈显著正相关(R2=0.23,P0.01),而它们均与土壤有机碳含量呈显著负相关。综上,土壤水分含量是影响紫色土自养微生物丰度和活性的关键因子,它们均随含水量升高而增加,此结果可为揭示土壤碳转化对土壤湿度变化的响应机制提供重要的基础信息。     

贺兰山东麓荒漠藻结皮微生物群落结构及其演替研究

以宁夏贺兰山东麓荒漠藻结皮为研究对象,对处于不同发育阶段的藻结皮中微生物群落结构及其演替进行了研究。结皮样品高通量测序结果分别得到521个16S rDNA序列操作分类单元(OTU)和64个18S rDNA序列OTU,表明藻结皮中原核微生物多样性远高于真核微生物;贺兰山东麓藻结皮中原核微生物分布于26个纲,Cyanobacteria在各个发育阶段中都是优势微生物类群,Actinobacteria、Chloroplast、Alphaproteobacteria和Bacilli在藻结皮发育的各个阶段相对丰度也较高;从属水平上分析,Bacillus、Leptolyngbya、Microcoleus、Microvirga、Chroococcidiopsis、Rubellimicrobium、Phormidium、Mastigocladopsis、Skermanella、Nostoc、Scytonema共11个属在各个发育阶段的藻结皮中都存在,只是出现了丰度的差异。Bacillus在藻结皮形成期、发育初期和发育中期相对丰度较大,成熟期丰度显著下降,而成熟期Microvirga丰度较前3个时期显著增加,表现出明显的细菌菌群演替现象。藻结皮样品中主要真核微生物分布于13个纲,其中Dothideomycetes和Pezizomycetes在各个发育阶段样品中丰度都很高,Agaricomycetes在形成期样品中相对丰度高达32.6%,但在藻结皮发育过程中其相对丰度迅速下降;原生生物的相对丰度在藻结皮发育过程中逐渐增加;4个发育阶段藻结皮样品中均未检测到真核微藻序列。4个发育阶段的藻结皮样品中有明确分类学信息的真核微生物共13个属,其中4个发育阶段中共有的为Coniothyrium、Dendryphion、Friedmanniomyces、Phloeopeccania、Sarocladium共5个属,其余8个属只在个别发育阶段样品中检出,表明在藻结皮发育过程中真核微生物的群落结构也发生着变化。藻结皮厚度、全氮含量及有机质含量是影响结皮层微生物群落组成的主要因素。     

混合电子供体促进非光合微生物菌群固碳效率的分子生态机制

前期研究已从全国各地的表层海水中分离驯化得到了能有效固碳的非光合固碳微生物菌群(Non-photosynthetic microbial community,NPMC),并发现在好氧条件下,混合电子供体(Mixture of Electron Donors,MEDs)(配比为0.46%NaNO_2、0.50%Na_2S_2O_3和1.25%Na_2S)能显著提高其固碳效率。但其与最佳H_2作为电子供体相比促进效果如何,其促进机制是什么,尚未见有报道。本文在比较了NPMC以MED和H_2为电子供体的固碳效率基础上,从NPMC的固碳基因丰度和表达活性角度分析了MED促进NPMC固碳效率的分子生态学机制。结果表明:(1) MED系统的固碳效率可以达到H_2系统的水平,并显著高于传统的NH_4~+系统的固碳效率;(2)卡尔文循环关键基因cbbL和cbbM在MED系统中的较高基因丰度和同时较高的表达可能是MED促进NPMC固碳效率的一个重要原因;(3)由于MED系统含有不同价态的N和S,有利于多种微生物生长,丰富了微生物多样性,为形成不同菌种之间的相互作用促进cbbL和cbbM的同时高表达提供了条件。     

长期施肥对稻田土壤固碳功能菌群落结构和数量的影响

微生物固碳在减缓全球气候变化、实现人类可持续发展方面具有重要的意义,通过揭示长期不同施肥制度对土壤固碳细菌的影响规律,可以为我国稻田土壤科学施肥,稻田固碳和温室气体减排的共轭双赢作用提供重要的理论依据。以湖南宁乡国家级稻田肥力变化长期定位试验为平台,采用PCR-克隆测序和实时荧光定量(Real-time)PCR技术,研究不施肥(CK),氮磷钾肥（NPK）和秸秆还田（NPKS）3种长期施肥制度对稻田土壤固碳细菌群落结构及数量的影响。通过分析固碳细菌cbbL基因文库发现,长期施肥导致土壤固碳细菌种群结构产生了明显差异,NPK和NPKS处理中兼性自养固碳菌群落优势增加而严格自养固碳菌生长受到抑制。LUBSHUFF软件统计分析显示cbbL基因文库在CK、NPK及NPKS处理间均存在显著性差异。 3种施肥处理的稻田土壤细菌cbbL基因拷贝数为3.35?108 —5.61?108每克土,施肥后,土壤细菌cbbL基因数量增加,其中NPKS处理cbbL数量最多,是CK处理的1.5倍左右。稀疏曲线则显示长期施化肥导致细菌cbbL基因多样性高于NPKS,而NPKS高于CK。上述结果表明了长期施肥对土壤固碳细菌群落结构,多样性及数量均有显著的影响。本研究结果可为深入探讨稻田土壤微生物固碳潜力及其影响机理提供有力的依据。     

金河湾城市湿地浮游植物功能类群演替及驱动因子

于2015年春(5月)、夏(8月)、秋(10月)三季,在金河湾湿地4类水体共设置12个采样点对浮游植物功能类群演替及与水环境变量关系进行分析。研究期间共鉴定浮游植物376个种,隶属于8门10纲19目19科101属。浮游植物种类组成主要以硅藻门(39.62%)和绿藻门(35.64%)为主,其次裸藻门(10.11%)和蓝藻门(9.84%)的藻类所占比例较高,甲藻门、隐藻门、金藻门和黄藻门所占比例较低。调查期间金河湾湿地浮游植物细胞丰度季节间差异显著(P0.05),整体上呈现夏季秋季春季的规律。春、夏、秋三季共划分20个不同的功能类群,双因素方差分析(Two-way ANOVA)和单因子交叉相似性检验(One-way ANOSIM)表明:代表性功能类群在季节间演替明显(P0.05),群落构成差异显著(P0.01)。SIMPER分析指出,S2/H1/B/D/Lo/X1/MP是引起金河湾湿地各季节之间浮游植物群落结构差异的主要贡献类群。通过代表性功能类群和10个水环境变量的典范对应分析(CCA)探索环境变量与功能类群演替的关系。经分析,总氮(TN)是驱动金河湾湿地浮游植物功能类群演替的主要环境变量,电导率(SpCond)、pH与功能类群演替密切相关。     

大青山不同树种根围土壤酶活性与微生物群落丰度的研究

针对大青山山地森林不同树种的根围土壤,探讨微生物丰度与土壤酶活性之间的联系以及受控因子。利用荧光实时定量PCR方法研究不同树种土壤微生物丰度的变化情况,分析土壤β-葡萄糖苷酶(βG)、N-乙酰氨基葡萄糖苷酶(NAG)和过氧化物酶(Pod)活性以及土壤理化性质的变化趋势。采用主成分分析(PCA)和皮尔森相关性分析方法研究土壤理化因子对土壤微生物群落丰度的影响。结果显示,土壤理化性质和微生物细菌、真菌群落丰度从春季到秋季均显著增高,如有机碳、全氮、微生物量碳和氮数量及细菌、真菌丰度等;大青山森林土壤βG、Pod酶活性夏季较高,而NAG酶活性秋季较高。皮尔森相关性分析表明大青山不同树种土壤酶活性与土壤微生物丰度有明显的相关性。与NAG酶活性极显著正相关(P0.05);与Pod酶活性呈极显著负相关(P0.05);而βG只与细菌群落丰度呈极显著负相关(P0.05)。土壤理化因子(有机碳、全氮、微生物量碳氮)与土壤微生物群落丰度均表现为极显著正相关。主成分分析认为,土壤有机碳、全氮、微生物量碳和氮、细菌和真菌群落丰度、N-乙酰氨基葡萄糖苷酶等可作为影响不同树种根围土壤养分特性的重要因子。在大青山山地森林生态系统,不同树种对土壤理化指标、土壤微生物丰度和土壤酶活性影响较大,并且随着季节变化响应较强。理化指标和土壤微生物群落丰度是调控大青山森林植被根围土壤微生物群落的主要生态因子。     

腾格里沙漠不同生物土壤结皮微生物多样性分析

主要采用MiSeq技术对腾格里沙漠不同生物土壤结皮中微生物16S rRNA基因V4-V5区进行测序, 分析其细菌群落结构的组成、丰度以及多样性, 通过非度量多维尺度法和Venn图解释微生物群落结构对生物土壤结皮演化过程的响应。结果表明不同生物土壤结皮在门水平上主要以Proteobacteria、Actinobacteria和Cyanobacteria为优势类群; 结皮下流沙层在门水平上主要以Actinobacteria、Proteobacteria、Chloroflexi和Acidobacteria为优势类群。在OTUs组成上, 样品之间存在共同的微生物类群, 也存在差异的微生物类群。结皮下流沙层的微生物群落结构组成相似, 与结皮中微生物群落结构的组成差异较大, 且藻结皮和藓结皮中微生物类群差异明显, 说明微生物类群对生物土壤结皮的演化过程做出了环境响应。在从流沙向藻结皮到藓结皮的演化过程中, 放线菌(Actinobacteria)、蓝细菌(Cyanobacteria)和变形菌(Proteobacteria)对结皮形成、演化的生物地球化学循环过程发挥着重要的影响。结皮中不同微生物类群丰度的差异表明其对沙漠土壤环境变化的响应以及生态功能的不同。     

玉米种植模式对褐土微生物特性的影响

土壤微生物群落组成及其酶活性是表明土壤肥力的重要敏感指标。本研究以辽西北褐土区设置的玉米等行距(对照)、二比空、三比空和大垄双行4种种植模式为对象,调查了玉米不同种植模式对褐土微生物特性的影响。结果表明:在4种种植模式的微生物群落组成中,细菌为优势类群,其中三比空、大垄双行磷脂脂肪酸总量、细菌、丛枝菌根真菌、革兰氏阳性细菌和革兰氏阴性细菌丰度均显著高于等行距和二比空模式;大垄双行种植模式土壤β-1,4-葡糖苷酶活性、腐生真菌丰度、微生物生物量碳和碱解氮含量均显著高于等行距种植模式,而酸性磷酸酶含量则显著低于等行距种植模式。相关分析表明:碱解氮含量与腐生真菌丰度呈极显著正相关(P0.01),与酸性磷酸酶活性呈显著负相关(P0.05)。大垄双行种植模式较高的腐生真菌丰度有利于分泌更多的β-1,4-葡糖苷酶,提高微生物生物量碳和碱解氮含量,进而促进碳氮循环过程,有利于玉米的生长。     

Composition of archaeal,bacterial, and eukaryal RuBisCO genotypes in three Western Pacific arc hydrothermal vent systems

We studied the diversity of all forms of the RuBisCO large subunit-encoding gene cbbL in three RuBisCO uncharacterized hydrothermal vent communities. This diversity included the archaeal cbbL and the forms IC and ID, which have not previously been studied in the deep-sea environment, in addition to the forms IA, IB and II. Vent plume sites were Fryer and Pika in the Mariana arc and the Suiyo Seamount, Izu-Bonin, Japan. The cbbL forms were PCR amplified from plume bulk microbial DNA and then cloned and sequenced. Archaeal cbbL was detected in the Mariana samples only. Both forms IA and II were amplified from all samples, while the form IC was amplified only from the Pika and Suiyo samples. Only the Suiyo sample showed amplification of the form ID. The form IB was not recorded in any sample. Based on rarefaction analysis, nucleotide diversity and average pairwise difference, the archaeal cbbL was the most diverse form in Mariana samples, while the bacterial form IA was the most diverse form in the Suiyo sample. Also, the Pika sample harbored the highest diversity of cbbL phylogenetic lineages. Based on pairwise reciprocal library comparisons, the Fryer and Pika archaeal cbbL libraries showed the most significant difference, while Pika and Suiyo showed the highest similarity for forms IA and II libraries. This suggested that the Fryer supported the most divergent sequences. All archaeal cbbL sequences formed unique phylogenetic lineages within the branches of anaerobic thermophilic archaea of the genera Pyrococcus, Archaeoglobus, and Methanococcus. The other cbbL forms formed novel phylogenetic clusters distinct from any recorded previously in other deep-sea habitats. This is the first evidence for the diversity of archaeal cbbL in environmental samples.     

纳木措可培养丝状真菌多样性及其与理化因子关系

微生物多样性在评估水体生态环境方面发挥着重要作用。本研究以青藏高原纳木措湖为研究对象, 开展水体可培养丝状真菌多样性及影响因子研究。通过膜过滤平置培养法、经典分类法和rRNA转录间隔区(ITS)序列分析对纳木措湖20个采样点的丝状真菌进行分离、纯化及鉴定, 测定水体理化指标, 综合分析丝状真菌空间分布格局与理化因子的相关性。菌种鉴定结果显示, 从纳木措水体样品中共分离纯化出1,412株丝状真菌, 隶属22属47种, 其中链格孢属(Alternaria)、青霉属(Penicillium)和毛霉属(Mucor)为优势属, 链格孢(Alternaria chlamydosporigena)和冻土毛霉(Mucor hiemalis)为优势种; Pearson相关性分析显示, 丝状真菌总丰度与温度、铵态氮、全磷呈显著正相关; 冗余分析显示, 铵态氮、温度、全磷、全氮、盐度及电导率是影响纳木措湖丝状真菌群落组成与分布的主要理化因子。综上所述, 纳木措水体可培养丝状真菌具有较高的物种多样性和空间异质性, 而且水体环境因子影响其分布。     

Hydroacoustic study of spatial and temporal distribution of Gymnocypris przewalskii (Kessler, 1876) in Qinghai Lake, China

Gymnocypris przewalskii (Kessler 1876) is an endangered and state-protected rare fish species in Qinghai Lake, China. To further understand the life history and distribution of this fish, five surveys were carried out in Qinghai Lake between 2002–2006. Results of these surveys indicate that fishes were predominantly distributed about 2 m under the surface. In July, significant differences in fish density were found between surface and bottom layers (P = 0.001), and/or between middle and bottom layers (P = 0.025). Fish density was the greatest in the surface layer. In August and October, no significant differences were found between the different layers, but the bottom layer had a greater fish density. Furthermore, there were very large differences among different zones in fish distribution density. Differences in horizontal distribution were not significantly correlated to factors such as water depth and inshore distance, possibly because of very low and uniform fish density. Feeding, changes in water temperature, over-wintering and spawning appeared to influence fish distribution. Hydroacoustic estimates of G. przewalskii biomass in Qinghai Lake increased significantly between 2002 and 2006. We attribute this increase to the management measures put in place to protect this species.     

Morphological variation of perch Perca fluviatilis in humic lakes: the effect of predator density,competition and prey abundance

Between and within‐lake variations in morphology of perch Perca fluviatilis were studied in four humic lakes in eastern Finland. Perca fluviatilis were more streamlined and smaller headed in a lake with the highest abundance of cyprinids, but lowest abundance of predators (Lake Tuopanjärvi), indicating adaptation to planktivorous feeding and low predator density. Highest bodied fish were found from a lake with the lowest cyprinid but highest predator abundance (Lake Koppelojärvi), which conversely indicates adaptation to more effective predator avoidance. Furthermore, the length of the paired fins was longest in Lake Kinnasjärvi and Lake Tuopanjärvi, where the abundance of benthic macroinvertebrates was lowest, suggesting selection for more effective benthivory. Clear morphological differences of P. fluviatilis between habitats were found only in Lake Kinnasjärvi, whereas in Lake Koppelojärvi and Lake Tuopanjärvi only the length of the paired fins differed and in Lake Harkkojärvi no differences were found. Taken together, these results suggest that inter and intrapopulation morphological differences are probably highly dependent on different biotic factors (i.e. predation risk, resource availability and competition). Spatial and temporal variations in these factors may have a great effect on body morphology of P. fluviatilis.     

Some studies on the ecology of limnetic crustacean zooplankton in Lake Begnas and Rupa,Pokhara Valley,Nepal

Monthly zooplankton samples taken during the period February, 1977 to February, 1978 in the deepest portion in two shallow sub-tropical Lakes, Begnas and Rupa in the Pokhara Valley, Nepal were studied. Four peaks of zooplankton abundance were noted. Adult and copepodid Copepoda were numerically dominant in both lakes with 56% Copepoda, 24% Cladocera and 20% nauplii in Lake Begnas and 48% Copepoda, 36% Cladocera and 16% nauplii in Lake Rupa. Other forms like Chaoborus larvae occurred sporadically in both lakes. An occurrence of the rare Limnocnida nepalensis (Coelenterata: Limnomedusae) in Lake Rupa was also noted during April and May, 1977. Although both of these lakes had already been ranked as eutrophic, the absence of calanoids, relative abundance of Bosmina longirostris and higher gross primary production in Lake Rupa is an indication of a higher trophic condition than that of Lake Begnas.     

Molecular Characterization of a Deep-Sea Methanotrophic Mussel Symbiont that Carries a RuBisCO Gene

In our previous investigation on the genes of 1,5-ribulose bisphosphate carboxylase/oxygenase (RuBisCO; EC 4.1.1.39) in deep-sea chemoautotrophic and methanotrophic endosymbioses, the gene encoding the large subunit of RuBisCO form I (cbbL) had been detected in the gill of a mussel belonging to the genus Bathymodiolus from a western Pacific back-arc hydrothermal vent. This study further examined the symbiont source of the RuBisCO cbbL gene along with the genes of 16S ribosomal RNA (16S rDNA) and particulate methane monooxygenase (EC 1.14.13.25; pmoA) and probed for the presence of the ATP sulfurylase gene (EC 2.7.7.4; sopT). The 16S rDNA sequence analysis indicated that the mussel harbors a monospecific methanotrophic Gammaproteobacterium. This was confirmed by amplification and sequencing of the methanotrophic pmoA, while thiotrophic sopT was not amplified from the same symbiotic genome DNA. Fluorescence in situ hybridization demonstrated simultaneous occurrence of the symbiont-specific 16S rDNA, cbbL and pmoA, but not sopT, in the mussel gill. This is the first molecular and visual evidence for a methanotrophic bacterial endosymbiont that bears the RuBisCO cbbL gene relevant to autotrophic CO₂ fixation.     

Effects of habitat fragmentation on genetic diversity and gene flow among the Homidia socia (Collembola: Entomobryidae) populations in the Thousand Island Lake

In the present study, partial sequences of the mitochondrial cytochrome oxidase subunit I (COI) gene of 22 island populations of the springtail Homidia socia in the Thousand Island Lake were sequenced. Across all sequences, 37 haplotypes were identified for the 510‐bp mitochondrial (mt) DNA COI gene. Haplotype 2 was the most common, and was distributed in the most of the 22 island populations. Haplotype diversity ranged from 0.065 to 0.733, and the total genetic diversity was 0.56216. The genetic characteristics of the 22 island populations were analyzed using the fixation index and gene flow, with values of 0.00043–0.94900 and 0.02703–703.72540, respectively. Comparison between (island area and isolations) with population genetic diversity revealed that there were no significant correlations between them, except for a significant correlation between the number of haplotypes and island area. Mantel tests showed that there was no significant correlation between geographic distance and genetic distance among various groups. All the results indicated that there were no obvious relationships between island characteristics and the genetic diversity of the springtails. We consider that the low dispersal capacity of springtails and the island patches surrounded by water in the Thousand Island Lake are the major factors affecting the genetic diversity of H. socia.