Forested headwater stream ecosystems are important integrators of terrestrial and aquatic systems and their function depends greatly on water availability. In the southern Appalachians, models of future climate change predict alterations to the timing and intensity of storms such that most precipitation may be relegated to winter and spring. During the summer and fall, relatively less precipitation will translate to lower stream flows in systems that rarely experience such a lack of water. Given these predicted changes to the hydrologic cycle, I experimentally reduced flow to downstream sections of three streams at the Coweeta Hydrologic Laboratory in NC to assess changes to function in perennial ecosystems. The questions that I addressed included: 1) How is organic matter decomposition regulated by changes to the availability of water? and 2) How does the relationship between nutrient uptake and metabolism change under conditions of varying water availability? The availability of water (as discharge) was shown to be a major control of ecosystem function throughout these studies. Rates of leaf decomposition varied between red maple (Acer rubrum L.) and white oak (Quercus alba L.) with lower discharge in the early autumn regulating the breakdown trajectories of leaves through facilitation of colonization by microbes and macroinvertebrates. The return of water during the winter accelerated decomposition rates in the diverted sites such that mass of leaves remaining were similar to those in upstream sections. Colonization of decomposing organic matter by heterotrophic microbes (especially fungi) increased N immobilization leading to an increase in respiration per unit leaf standing stocks during the fall. Nitrification was detectable during summer low flows when leaf standing stocks were low. Changes in the timing and intensity of precipitation and thus discharge may in turn alter the temporal dynamics of ecosystem function. Leaves may remain in the stream unprocessed which will change the availability of food for macroinvertebrates, the production of which provides nutrition to higher trophic levels. Local-scale differences in organic matter processing and nutrient immobilization may translate to regional differences in food availability over both time and space. Hydrology not only acts as a local control of endogenous processes but acts also regionally through the transport of resources and nutrients to downstream reaches. / Ph. D.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21312号 / 農博第2297号 / 新制||農||1065(附属図書館) / 学位論文||H30||N5146(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 舟川 晋也, 教授 縄田 栄治, 教授 北山 兼弘 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

I investigated the drivers of nutrient cycling by heterotrophic microbes during leaf decomposition in streams. My research addressed two overarching questions: 1) how do exogenous and endogenous factors interact to drive microbial nitrogen (N) cycling during organic matter decomposition in stream ecosystems, and 2) what affect will the global increase in biologically active N have on these factors and resulting fluxes? I conducted studies in natural streams and laboratory mesocosms to address these questions and used general stoichiometric theory to conceptualize diverse microbial assemblages as a single functional unit within stream ecosystems. First, I described spatial and temporal patterns of N and phosphorus uptake and mineralization by leaf-associated microbial assemblages in five southern Appalachian streams which spanned a gradient of nitrate availability. I found wide variations in nutrient fluxes across time and space, perhaps due to macroinvertebrate-induced changes in microbial assemblage composition. Secondly, I explored the roles of endogenous and exogenous N in meeting microbial requirements. I isolated microbial biomass from leaves that had been labeled with N-15 and incubated in the same five Appalachian streams. The importance of exogenous N increased as decomposition progressed and was particularly important in streams with high N availability. Finally, I tested potential interactions between two exogenous drivers of microbial nutrient cycling: N availability and animal activity. I used mesocosms to test the effects of consumer nutrient recycling (CNR) and grazing by two shredders on microbial uptake under different N regimes. Animals only influenced microbial uptake under low N conditions. Shredder CNR generally stimulated uptake while grazing had a negative effect. My research provides a robust model describing N cycling by detritus-associated microbes over the course of decomposition. According to this model, microbes assimilate endogenous N during the initial stages of decomposition and immobilization of exogenous N becomes more important as decomposition progresses. The labeled substrate technique that I used to generate this model is an elegant way of testing the applicability of this model in other ecosystems. My results also suggest that anthropogenic activities that increase exogenous N availability have implications for N and C cycling in lotic systems. / Ph. D.

Estudaram-se a estruturas taxionômica e trófica e determinaram-se os índices de maturidade de comunidades de nematóides ocorrentes em áreas contíguas de mata natural preservada e de culturas perenes (bananeira, citros, pessegueiro) e anuais (milho x leguminosas) em rotação em Piracicaba (SP). Três amostras compostas de solo (volume aproximado de um litro) foram coletadas de cada área em duas épocas (\"seca\", agosto/2004 e \"chuvosa\", março/2005) e profundidades (0-15 e 15-30 cm). Para cada amostra, extraíram-se os nematóides de volume de 500 ml pela técnica de peneiramento combinado com flotação em centrífuga com solução de sacarose, sendo depois fixados em formalina, contados e infiltrados com glicerina. Para o cálculo das abundâncias, indicadores de diversidade e os índices de maturidade, 150 exemplares foram coletados ao acaso por amostra e identificados ao nível de gênero. Assinalaramse 61 gêneros, filiados a 32 famílias. As abundâncias totais foram maiores à menor profundidade, nas duas épocas de coleta. Os maiores valores de número e riqueza de gêneros, à menor profundidade, ocorreram na mata; à maior profundidade, tal se deu na área de anuais e de citros, nas primeira e segunda épocas de coleta, respectivamente. Os índices de diversidade de Shannon- Weaver e de Simpson e respectivos índices de eqüitatividade foram menos eficientes que o número e a riqueza de gêneros na discriminação dos sistemas estudados. Os parasitos de plantas constituíram o grupo trófico prevalente em todos os sistemas avaliados, nas duas épocas e profundidades de coleta, seguidos no geral pelos bacteriófagos. Todavia, não foi possível separar cultivos anuais de perenes com base nas abundâncias de parasitos de plantas. Os valores de T (diversidade trófica) não se prestaram à clara distinção entre os cinco sistemas, mas os de MI, mMI e PPI indicaram menor ocorrência de distúrbios na mata e no pomar cítrico em relação às demais áreas cultivadas. A herbivoria foi o canal prevalente de decomposição nos diferentes sistemas, exceto no pomar de citros. Dentre os microbiófagos (FF/BF), a predominância foi da via bacteriana. / The taxonomic structure, trophic structure and maturity index of nematode communities occurring in neighbouring areas of native forest and four different agroecosystems were studied in Piracicaba, São Paulo State, Brazil. Agroecosystems were three perennial (banana, citrus and peach orchards) and one annual (corn, rotating with leguminous plants or followed by fallow) crops. Three soil samples (one liter approximately) were collected per area at two different times (August/2004 and March/2005) and depths (0-15 and 15-30 cm). Nematodes were extracted from 500 cc of soil by a sieving and sugar flotation technique, fixed with formalin, counted and infiltrated with glycerin. Subsequently, 150 specimens randomly picked were identified to the genus level. A total number of 61 genera affiliated to 32 families was recorded. Total abundance was higher at lower depth irrespective of the sampling time. At depth 0-15 cm, higher values of genus number and genus richness were calculated for the forest in both sampling times; at 15-30 cm, the highest values were assigned to annual crop and citrus areas, respectively. Shannon- Weaver and Simpson indexes were less efficient than genus number and genus richness in discriminating the studied ecosystems. Plant-parasitic nematodes prevailed as a trophic group in all systems at both sampling times and depths, the bacterivorous usually ranking second position. However, the data of relative abundance of phytonematodes did not allow annual and perennial crop areas to be statistically differentiated. The studied systems also were not clearly distinguished with basis on the T values, but according to the MI, mMI and PPI data the forest and the citrus orchard were the less disturbed areas. Herbivory was the predominant decomposition channel in all systems, with exception of the citrus orchard. In terms of microbial decomposition (FF/BF), the process was usually dominated by bacteria.

La lignine est un des polymères naturels les plus abondants sur la terre particulièrement résistant à la dégradation du fait de sa structure particulière qui lui permet de participer au renforcement des parois végétales ainsi qu'à la protection de celles-ci contre l'attaque de pathogènes. Peu d'organismes peuvent dégrader efficacement la lignine et la plupart de ceux-ci appartiennent au Règne Fongique. Dans la plupart des cas la dégradation des composants de la paroi végétale est réalisée par des enzymes sécrétées. Des études récentes soulignent combien l'arsenal enzymatique des champignons est intéressant dans plusieurs domaines :- nombreuses de ces enzymes sont déjà utilisées en biotechnologies dans de nombreux et divers processus industriels,- grâce à la possibilité de comparer les génomes de nombreuses espèces fongiques, au travers de l'étude des gènes impliqués dans la matière organique il apparait que l'évolution fongique et l'écologie de ces microorganismes sont étroitement liés,- les champignons dégradateurs de la matière organique végétale sont les principaux décomposeurs en milieu terrestre. Ils contribuent notamment de manière décisive dans le cycle biogéochimique du carbone en milieu forestier tempéré et boréal. L'étude et une meilleure compréhension des flux de carbone est importante pour mieux appréhender les phénomènes de changements climatiques.L'appareil enzymatique fongique permettant la dégradation de la matière organique est composé de dizaines, sinon de centaines d'enzymes aux rôles bien décrits pour certaines. Mais pour d'autres, leurs rôles restent à définir. C'est le cas des peroxydases de type DyP, une famille enzymatique découverte récemment et déjà bien connue dans le domaine des biotechnologies, étant capables de catalyser de nombreuses réactions. Leurs rôles dans la nature sont toutefois incertains. Il a été suggéré qu'elles puissent jouer un rôle dans la dégradation de la lignine, mais certaines études suggèrent plutôt un rôle de détoxification durant la dégradation de la biomasse.Au cours de cette thèse, nous nous sommes attachés à mettre en lumière les rôles potentiels joués par ces enzymes. La famille des DyP a récemment été divisée en sous-groupes, mais pour ce qui concerne les DyP fongiques, il n'existait pas d'analyse approfondie de leur évolution phylogénique. Un tel travail nous a conduit à mettre en lumière l'existence ignorée jusqu'à présent de DyP aussi bien intracellulaires qu'extracellulaires.Afin d'améliorer nos connaissances sur cette famille enzymatique nous nous sommes orientés sur des analyses écologiques pour lesquelles nous avons dû au préalable développer des outils appropriés tel qu'un protocole d'extraction d'ARN à partir de bois en décomposition.Par une approche d'écologie moléculaire, nous avons évalué quelle est la source et la diversité des champignons producteurs de DyP dans 3 habitats distincts : des sols forestiers, des sols de prairies et du bois en décomposition.Une analyse par métabarcoding des communautés fongiques de ces environnements a au préalable été réalisée et a démontré le bénéfice que l'on peut tirer en métabarcoding de l'analyse comparative de l'ADN et de l'ARN environnemental pour une description optimale de ces communautés.L'étude de la diversité des DyP exprimées au sein des communautés fongiques présentes dans ces différents habitats a été réalisée par capture de séquences à partir d'ARN environnementaux.Les résultats préliminaires obtenus démontrent la capacité de l'approche expérimentale à isoler les gènes pleine longueur correspondants de tous les échantillons étudiés. Certaines de ces DyP environnementales ont été transformées dans le champignon Podospora anserina, et l'expression de l'une d'entre elle dans ce système hétérologue a pu être démontrée [etc...] / Lignin is one of the most abundant natural polymers on earth whose resistance to degradation contributes to the mechanical strength of plant cellwalls and protects plant cells from pathogen attack. Few organisms are candegrade lignin efficiently; most of them belong to the fungal Kingdom. Inmost cases, plant cell wall degradation results from enzymatic attack. Recent studies also underline the contribution of secreted fungal enzymes in many areas:- many of these enzymes are already in use in numerous industrial processes,- thanks to the comparison between fungal genomes, through the study ofgenes implicated in organic matter degradation, it becomes obvious that fungal evolution and their ecology are two tightly linked phenomena,- saprotrophic fungi degrading organic matter are the main decomposers in terrestrial ecosystems. Among other, they contribute in a decisive way tothe carbon biogeochemical cycle in temperate and boreal ecosystems. Thestudy and a better comprehension of carbon fluxes are of prime importance in the evaluation of climate changes.The fungal enzymatic machinery involved in organic matter decomposition is compozed of dozens of enzymes whose functions are diversely understood. The roles of several of them need to be clarified. This is the case for the DyP peroxidases, an enzyme family recently described but already well known in the field of biotechnology for their capacity of catalyzing many reactions. Their natural role in natural ecosystems are however matter of discussion. It has been suggested that they could participate in lignin degradation although a role in detoxification during biomass degradation cannot be excluded.In the course of this thesis, we highlighted the potential roles of these enzymes.The DyP gene family had been divided in different sub-families but nostudy specifically dealt with the phylogeny of fungal DyPs. Such an analysis revealed the unsuspected existence of both intracellular and extracellular DyPs in fungi. To better understand the potential roles of this fungal gene family we developed ecological analyses that first required the development of specific tools such as a protocol to extract RNA directly from decomposing wood. Following a molecular ecology approach, we evaluated the source and diversity of DyP-producing fungi in three distinct habitats; grassland soils, forest soils and decomposing wood. A metabarcoding analysis of the fungal communities present in these different environments has first been conducted and has revealed the beneficial impact of performing metabarcoding on both environmental DNA and RNA to accurately describe fungal communities.The study of DyPs expressed within fungal communities colonizing these different habitats has been conducted by sequence capture on environmental RNA. Preliminary results demonstrate the validity of this approach to isolate the corresponding full-length genes from all studies environmental samples. Several of these environmental DyP genes were transformed in the fungus Podospora anserina and the expression of one of them in this heterologous host was demonstrated.In conclusion, DyP peroxidases still represent a family of fungal enzymes of unclear role. We suggest that extracellular and intracellular DyPs may play complementary roles in both lignin degradation and detoxification of toxic environmental compounds, respectively. This enzyme family is more specifically present in the genomes of basidiomycete fungi capable of enzymatic deconstrustion of lignin. A restricted number of DyP genes has been isolated from each of the different studied environmental samples, thus suggesting that the corresponding enzymes are not abundantly produced although present in all environments

Estudaram-se a estruturas taxionômica e trófica e determinaram-se os índices de maturidade de comunidades de nematóides ocorrentes em áreas contíguas de mata natural preservada e de culturas perenes (bananeira, citros, pessegueiro) e anuais (milho x leguminosas) em rotação em Piracicaba (SP). Três amostras compostas de solo (volume aproximado de um litro) foram coletadas de cada área em duas épocas (\"seca\", agosto/2004 e \"chuvosa\", março/2005) e profundidades (0-15 e 15-30 cm). Para cada amostra, extraíram-se os nematóides de volume de 500 ml pela técnica de peneiramento combinado com flotação em centrífuga com solução de sacarose, sendo depois fixados em formalina, contados e infiltrados com glicerina. Para o cálculo das abundâncias, indicadores de diversidade e os índices de maturidade, 150 exemplares foram coletados ao acaso por amostra e identificados ao nível de gênero. Assinalaramse 61 gêneros, filiados a 32 famílias. As abundâncias totais foram maiores à menor profundidade, nas duas épocas de coleta. Os maiores valores de número e riqueza de gêneros, à menor profundidade, ocorreram na mata; à maior profundidade, tal se deu na área de anuais e de citros, nas primeira e segunda épocas de coleta, respectivamente. Os índices de diversidade de Shannon- Weaver e de Simpson e respectivos índices de eqüitatividade foram menos eficientes que o número e a riqueza de gêneros na discriminação dos sistemas estudados. Os parasitos de plantas constituíram o grupo trófico prevalente em todos os sistemas avaliados, nas duas épocas e profundidades de coleta, seguidos no geral pelos bacteriófagos. Todavia, não foi possível separar cultivos anuais de perenes com base nas abundâncias de parasitos de plantas. Os valores de T (diversidade trófica) não se prestaram à clara distinção entre os cinco sistemas, mas os de MI, mMI e PPI indicaram menor ocorrência de distúrbios na mata e no pomar cítrico em relação às demais áreas cultivadas. A herbivoria foi o canal prevalente de decomposição nos diferentes sistemas, exceto no pomar de citros. Dentre os microbiófagos (FF/BF), a predominância foi da via bacteriana. / The taxonomic structure, trophic structure and maturity index of nematode communities occurring in neighbouring areas of native forest and four different agroecosystems were studied in Piracicaba, São Paulo State, Brazil. Agroecosystems were three perennial (banana, citrus and peach orchards) and one annual (corn, rotating with leguminous plants or followed by fallow) crops. Three soil samples (one liter approximately) were collected per area at two different times (August/2004 and March/2005) and depths (0-15 and 15-30 cm). Nematodes were extracted from 500 cc of soil by a sieving and sugar flotation technique, fixed with formalin, counted and infiltrated with glycerin. Subsequently, 150 specimens randomly picked were identified to the genus level. A total number of 61 genera affiliated to 32 families was recorded. Total abundance was higher at lower depth irrespective of the sampling time. At depth 0-15 cm, higher values of genus number and genus richness were calculated for the forest in both sampling times; at 15-30 cm, the highest values were assigned to annual crop and citrus areas, respectively. Shannon- Weaver and Simpson indexes were less efficient than genus number and genus richness in discriminating the studied ecosystems. Plant-parasitic nematodes prevailed as a trophic group in all systems at both sampling times and depths, the bacterivorous usually ranking second position. However, the data of relative abundance of phytonematodes did not allow annual and perennial crop areas to be statistically differentiated. The studied systems also were not clearly distinguished with basis on the T values, but according to the MI, mMI and PPI data the forest and the citrus orchard were the less disturbed areas. Herbivory was the predominant decomposition channel in all systems, with exception of the citrus orchard. In terms of microbial decomposition (FF/BF), the process was usually dominated by bacteria.

No description available.

第23回名古屋大学年代測定総合研究センターシンポジウム平成22(2010)年度報告

Dead wood is one of the most important reservoirs associated with forest ecosystems. In natural forests, its volume is counted in hundreds of m3 ha-1 , whereas it reaches only tens of m3 ha-1 in productive commercial forests. In contrast to soil and plant litter, deadwood is unevenly distributed on the forest floor. The specific physicochemical properties such as high content of recalcitrant polymers, low nitrogen level and impermeability negatively affect the rate of decomposition especially in the initial stages of wood deconstruction. The deadwood decomposition is very slow in comparisons with other substrates, it accumulates and thus it represents the important reservoir of nutrients. This thesis is focused on the structure, development and function of microbial (fungal) community in decomposing deadwood in unmanaged forest. Functional screening of fungi isolated from fruit bodies collected from coarse deadwood was set aside. Physico-chemical properties of deadwood including pH, carbon and nitrogen content and microbial biomass were estimated for four wood decomposition stages and three different tree species. New generation sequencing (Illumina MiSeq platform) was applied for fungal community structure analysis based on ITS2 fragment. Fungal functional screening was based on physico-chemical...

The objective of the thesis is the model-based analysis of spatial patterns of decomposition properties on the forested slopes of the montane level (ca. 1200-2200 m a.s.l.) in a study area in the Italian Alps (Val di Sole / Val di Rabbi, Autonomous Province of Trento). The analysis includes humus forms and enchytraeid assemblages as well as pH values, activities of extracellular enzymes and C/N ratios of the topsoil. The first aim is to develop, test and apply data-based techniques for spatial modelling of soil ecological parameters. This methodological approach is based on the concept of digital soil mapping. The second aim is to reveal the relationships between humus forms, soil organisms and soil microbiological parameters in the study area. The third aim is to analyze if the spatial patterns of indicators of decomposition differ between the landscape scale and the slope scale. At the landscape scale, sample data from six sites are used, covering three elevation levels at both north- and south-facing slopes. A knowledge-based approach that combines a decision tree analysis with the construction of fuzzy membership functions is introduced for spatial modelling. According to the sampling design, elevation and slope exposure are the explanatory variables. The investigations at the slope scale refer to one north-facing and one south-facing slope, with 30 sites occurring on each slope. These sites have been derived using conditioned Latin Hypercube Sampling, and thus reasonably represent the environmental conditions within the study area. Predictive maps have been produced in a purely data-based approach with random forests. At both scales, the models indicate a high variability of spatial decomposition patterns depending on the elevation and the slope exposure. In general, sites at high elevation on north-facing slopes almost exclusively exhibit the humus forms Moder and Mor. Sites on south-facing slopes and at low elevation exhibit also Mull and Amphimull. The predictions of those enchytraeid species characterized as Mull and Moder indicators match the occurrence of the corresponding humus forms well. Furthermore, referencing the mineral topsoil, the predictive models show increasing pH values, an increasing leucine-aminopeptidase activity, an increasing ratio alkaline/acid phosphomonoesterase activity and a decreasing C/N ratio from north-facing to south-facing slopes and from high to low elevation. The predicted spatial patterns of indicators of decomposition are basically similar at both scales. However, the patterns are predicted in more detail at the slope scale because of a larger data basis and a higher spatial precision of the environmental covariates. These factors enable the observation of additional correlations between the spatial patterns of indicators of decomposition and environmental influences, for example slope angle and curvature. Both the corresponding results and broad model evaluations have shown that the applied methods are generally suitable for modelling spatial patterns of indicators of decomposition in a heterogeneous high mountain environment. The overall results suggest that the humus form can be used as indicator of organic matter decomposition processes in the investigated high mountain area.