Variable use of plant- and soil-derived carbon by microorganisms in agricultural soils

In this study we used compound specific ¹³C and ¹⁴C isotopic signatures to determine the degree to which recent plant material and older soil organic matter (SOM) served as carbon substrates for microorganisms in soils. We determined the degree to which plant-derived carbon was used as a substrate by comparison of the ¹³C content of microbial phospholipid fatty acids (PLFA) from soils of two sites that had undergone a vegetation change from C3 to C4 plants in the past 20-30 years. The importance of much older SOM as a substrate was determined by comparison of the radiocarbon content of PLFA from soils of two sites that had different ¹⁴C concentrations of SOM.The ¹³C shift in PLFA from the two sites that had experienced different vegetation history indicated that 40-90% of the PLFA carbon had been fixed since the vegetation change took place. Thus PLFA were more enriched in ¹³C from the new C4 vegetation than it was observed for bulk SOM indicating recent plant material as preferentially used substrate for soil microorganisms. The largest ¹³C shift of PLFA was observed in the soil that had high ¹⁴C concentrations of bulk SOM. These results reinforce that organic carbon in this soil for the most part cycles rapidly. The degree to which SOM is incorporated into microbial PLFA was determined by the difference in ¹⁴C concentration of PLFA derived from two soils one with high ¹⁴C concentrations of bulk SOM and one with low. These results showed that 0-40% of SOM carbon is used as substrate for soil microorganisms. Furthermore a different substrate usage was identified for different microorganisms. Gram-negative bacteria were found to prefer recent plant material as microbial carbon source while Gram-positive bacteria use substantial amounts of SOM carbon. This was indicated by ¹³C as well as ¹⁴C signatures of their PLFA. Our results find evidence to support ‘priming’ in that PLFA indicative of Gram-negative bacteria associated with roots contain both plant- and SOM-derived C. Most interestingly, we find PLFA indicative of archeobacteria (methanothrophs) that may indicate the use of other carbon sources than plant material and SOM to a substantial amount suggesting that inert or slow carbon pools are not essential to explain carbon dynamics in soil.     

Identification of active bacteria involved in decomposition of complex maize and soybean residues in a tropical Vertisol using N-DNA stable isotope probing

As limited information is available about the relationship between microbial processes and community structure in tropical soils, we used ¹⁵N-DNA stable isotope probing (¹⁵N-DNA-SIP) to identify bacteria actively involved in decomposition of plant residues of different biochemical quality. ¹⁵N-labeled (90 atom%) and unlabeled (control) maize (C-to-N ratio: 32; cellulose content: 24.9%) and soybean (15; 15.5%) leaf residues were incubated in a tropical Vertisol for 15 days. Soil DNA was isolated, subjected to ¹⁵N-DNA-SIP and buoyant density-resolved DNA fractions were analyzed by 16S rRNA gene-based denaturing gradient gel electrophoresis (DGGE) analysis and sequencing of selected DGGE bands. Residue addition induced new bands and changed relative intensity of already existing bands in ¹⁵N-enriched SIP fractions. Phylogenetic analysis of selected, cloned DGGE bands from ‘heaviest’ ¹⁵N-enriched fractions (57.8 atom% (maize), 87.1 atom% (soybean)) revealed that soils treated with maize residues were dominated by Pseudonocardia sp., while Arthrobacter sp. and Streptomyces sp. were found in the soybean residue treated soils. Sequences related to Bacillus sp. and Saccharopolyspora sp. were found in both organic residue treatments. Our study gave clear evidence that ¹⁵N-DNA-SIP combined with 16S rRNA gene-based community fingerprinting of density-resolved fractions and an unlabeled control was suited for detecting active bacteria involved in decomposition of complex maize and soybean residues. In conclusion, we could show that residue quality, inducing contrasting N assimilation by decomposing bacteria, was a substantial determinant of certain decomposing community members assayed in this study.     

Effects of food quality, starvation and life stage on stable isotope fractionation in Collembola

Naturally occurring stable isotopes of carbon and nitrogen are powerful tools to investigate food webs, where the ratio of ¹⁵N/¹⁴N is used to assign trophic levels and of ¹³C/¹²C to determine the food source. A shift in δ¹⁵N value of 3‰ is generally suggested as mean difference between two trophic levels, whereas the carbon isotope composition of a consumer is assumed to reflect the signal of its diet. This study investigates the effects of food quality, starvation and life stage on the stable isotope fractionation in fungal feeding Collembola. The fractionation of nitrogen was strongly affected by food quality, i.e. the C/N ratio of the fungal diet. Collembola showed enrichment in the heavier isotope with increasing N concentration of the food source. Δ¹⁵N varied between 2.4‰, which assigns a shift in one trophic level, and 6.3‰, suggesting a shift in two trophic levels. Starvation up to 4 weeks resulted in an increase in the total δ¹⁵N value from 2.8‰ to 4.0‰. Different life stages significantly affected the isotope discrimination by Collembola with juveniles showing a stronger enrichment (Δ¹⁵N=4.9‰) compared to adults (Δ¹⁵N=3.5‰). Δ¹³C varied between −2.1‰ and −3.3‰ depending on the food quality, mainly due to compensational feeding on low quality diet. During starvation δ¹³C value decreased by 1.1‰, whereas the life stage of Collembola had no significant effect on isotopic ratios. The results indicate that the food resource and the physiological status of the consumer have important impact on stable isotope discrimination. They may cause differences in fractionation rate comparable to trophic level shifts, a fact to consider when analysing food web structure.     

基于稳定性同位素核酸探针技术的红壤微生物底物利用策略研究

外源活性碳底物输入强烈影响土壤微生物的生长,但是在系统发育分类水平上,细菌和真菌对活性底物的动态响应及利用特征和微生物群落组成的关系仍不清楚。以红壤为研究对象,采用¹³C标记葡萄糖为底物进行模拟培养并定期取样,利用稳定性同位素核酸探针(DNA-based stable isotope probing,DNA-SIP)和高通量测序技术分析活性的真菌和细菌类群,并探讨不同微生物利用葡萄糖来源碳的动态特征。研究发现,微生物群落对葡萄糖的利用符合从细菌向真菌演替的r/K选择策略。在细菌群落中,隶属于富营养菌的变形菌门(Proteobacteria)和放线菌门(Actinobacteria)对活性底物的利用能力显著高于寡营养菌的酸杆菌门(Acidobacteria)和绿弯菌门(Chloroflexi)。和细菌的底物利用策略不同,真菌子囊菌门(Ascomycota)和担子菌门(Basidiomycota)在整个培养期间均可以利用葡萄糖和土壤原有组分。因此,葡萄糖的连续加入并未改变不同营养类型细菌的底物利用策略,而真菌对底物的利用具有广谱性特征,活性底物可诱导真菌对土壤原有组分...     

Feeding guilds in Collembola based on nitrogen stable isotope ratios

In soil a high number of species co-exist without extensive niche differentiation, which was assigned as ‘the enigma of soil animal species diversity’. In particular, the detritivores are regarded as food generalists. We have investigated nitrogen stable isotope ratios (¹⁵N/¹⁴N) of a major decomposer group, the Collembola, to evaluate trophic relationship and determine feeding guilds. Additionally, the δ¹⁵N values of potential food sources such as mosses, lichens and other plant derived material (bark, nuts, leaves) were analysed. The natural variation in nitrogen isotopes was assessed in 20 Collembola taxa from three deciduous forest stands. The δ¹⁵N signature formed a continuum from phycophages/herbivores to primary and secondary decomposers, reflecting a gradual shift from more detrital to more microbial diets. The δ¹⁵N gradient spanned over 9 δ units, which implies a wide range in food sources used. Assuming a shift in ¹⁵N of about 3 ‰ per trophic level, the results indicate a range of three trophic levels. These variations in ¹⁵N/¹⁴N ratios suggest that trophic niches of Collembola species differ and this likely contributes to Collembola species diversity.     

Identification of in situ 2,4-dichlorophenoxyacetic acid-degrading soil microorganisms using DNA-stable isotope probing

Stable isotope probing (SIP) was used to investigate the microorganisms responsible for degradation of the herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D) in soil samples. Soils were unamended or amended with either unlabeled 2,4-D or UL(ring) ¹³C-2,4-D. Degradation of 2,4-D was complete after 17 days, whereas little removal (11±3%) was observed in the sterile controls. Terminal restriction fragment length polymorphism (TRFLP) on soil DNA after 17 days indicated a consistent increase in the relative abundance of one fragment (217 bp in Hae III digests) in soils spiked with 2,4-D (both unlabeled and labeled samples) compared to the unamended soils. DNA extracts from labeled and unlabeled 2,4-D amended soils were subject to ultracentrifugation, fractionation of centrifuged samples, followed by TRFLP on each fraction. TRFLP profiles from ultracentrifugation fractions illustrated that the same fragment experienced an increase in buoyant density (BD) in samples spiked with ¹³C-labeled 2,4-D. This increase in DNA BD indicates the organisms represented by this fragment were responsible for uptake and degradation of the herbicide. 16S rRNA sequencing of the heavy, ¹³C-enriched fraction suggests the organisms belong to the β subdivision of Proteobacteria. Herein, SIP facilitated the identification of unique organisms degrading 2,4-D in soil without the need for isolation and provided more direct evidence for a functional role of these organisms than would have been possible with the molecular-based methods alone.     

Microbial immobilisation of C rhizodeposits in rhizosphere and root-free soil under continuous C labelling of oats

A greenhouse experiment was conducted by growing oats (Avenasativa L.) in a continuously ¹³CO₂ labeled atmosphere. The allocation of ¹³C-labeled photosynthates in plants, microbial biomass in rhizosphere and root-free soil, pools of soil organic C, and CO₂ emissions were examined over the plant's life cycle. To isolate rhizosphere from root-free soil, plant seedlings were placed into bags made of nylon monofilament screen tissue (16 μm mesh) filled with soil. Two peaks of ¹³C in rhizosphere pools of microbial biomass and dissolved organic carbon (DOC), as well as in CO₂ emissions at the earing and ripeness stages were revealed. These ¹³C maxima corresponded to: (i) the end of rapid root growth and (ii) beginning of root decomposition, respectively. The δ¹³C values of microbial biomass were higher than those of DOC and of soil organic matter (SOM). The microbial biomass C accounted for up to 56 and 39% of ¹³C recovered in the rhizosphere and root-free soil, respectively. Between 4 and 28% of ¹³C assimilated was recovered in the root-free soil. Depending on the phenological stage, the contribution of root-derived C to total CO₂ emission from soil varied from 61 to 92% of total CO₂ evolved, including 4-23% attributed to rhizomicrobial respiration. While 81-91% of C substrates used for microbial growth in the root-free soil and rhizosphere came from SOM, the remaining 9-19% of C substrates utilized by the microbial biomass was attributable to rhizodeposition. The use of continuous isotopic labelling and physical separation of root-free and rhizosphere soil, combined with natural ¹³C abundance were effective in gaining new insight on soil and rhizosphere C-cycling.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.     

Microbial response to exudates in the rhizosphere of young beech trees (Fagus sylvatica L.) after dormancy

Plants act as an important link between atmosphere and soil: CO2 is transformed into carbohydrates by photosynthesis. These assimilates are distributed within the plant and translocated via roots into the rhizosphere and soil microorganisms. In this study, 3 year old European beech trees (Fagus sylvatica L.) were exposed after the chilling period to an enriched ¹³C–CO₂ atmosphere (δ¹³C = 60‰ – 80‰) at the time point when leaves development started. Temporal dynamics of assimilated carbon distribution in different plant parts, as well as into dissolved organic carbon and microbial communities in the rhizosphere and bulk soil have been investigated for a 20 days period. Photosynthetically fixed carbon could be traced into plant tissue, dissolved organic carbon and total microbial biomass, where it was utilized by different microbial communities. Due to carbon allocation into the rhizosphere, nutrient stress decreased; exudates were preferentially used by Gram-negative bacteria and (mycorrhizal) fungi, resulting in an enhanced growth. Other microorganisms, like Gram-positive bacteria and mainly micro eucaryotes benefited from the exudates via food web development. Overall our results indicate a fast turnover of exudates and the development of initial food web structures. Additionally a transport of assimilated carbon into bulk soil by (mycrorhizal) fungi was observed.     

Isotopic detection of recent photosynthate carbon flow into grassland rhizosphere fauna

In this study, we measured the incorporation of recent photosynthate-C inputs into active rhizosphere fauna (earthworms, enchytraeids, mites and collembolans) in an upland grassland soil under natural environmental conditions. This was achieved by means of a ¹³CO₂ pulse-chase experiment made during the growing season, followed by a 20-day dynamic sampling of soil fauna for ¹²C/¹³C analysis by IRMS. The effect of post-¹³C labelling defoliation (cutting) on fauna ¹²C/¹³C ratios was also examined.Results showed that earthworms made up over 93% of the extracted fauna biomass, while mites, collembolans and enchytraeids together accounted for less than 7%. All fauna groups showed evidence of tracer ¹³C in their tissues within a week of ¹³CO₂ pulse labelling in both control and cut treatments. Cutting significantly increased the amount of tracer ¹³C entering the organisms (P=0.0002). Similarly, the fauna group also had a significant effect (P=0.0001). Time did not have any effect on fauna ¹³C content between groups as differences were only significant at the last sampling occasion. The interaction time×animal group, however, had a significant effect (P=0.0054).Collembolans accounted for most of the tracer ¹³C measured within the fauna biomass, i.e. mean±standard deviation of 44.78±12.75% and 44.29±14.69% of fauna ¹³C in control and cut treatments, respectively. Mites and earthworms contained between 22.13% and 28.45%, and enchytraeids less than 6% of the tracer ¹³C. We conclude that, during the growing season, there was a rapid incorporation of recent photosynthate-C into rhizosphere mesofauna. This carbon transfer was most significantly increased by defoliation in mites and collembolans (P<0.01). These results provide evidence that soil foodweb carbon dynamics are not solely underpinned by detrital decomposition but are also affected by short-term plant rhizodeposition patterns.     

Separation of soil respiration into CO2 emission sources using 13C natural abundance in a deciduous broad-leaved forest in Japan

Abstract

In determining the soil and ecosystem carbon balance, it is necessary to distinguish between autotrophic respiration and heterotrophic respiration. We attempted to measure the contribution of CO₂ emissions from plant roots (R_RHI), from soil organic matter (R_SOM), and from litter (R_L) to CO₂ emissions from the forest floor (soil respiration; R_S) in a deciduous forest of oak (Quercus serrata Thunb.) and hornbeams (Carpinus laxiflora Sieb. et Zucc. Bl., Carpinus tschonoskii Maxim. and Carpinus japonica Bl.) on Andosols in Japan, using a ¹³C natural abundance technique. The ¹³C natural abundances of roots (δ_RHI), litter (δ_L) and SOM (δ_SOM) in the surface soil were ?28.9, ?30.1 and ?24.3‰, respectively. This means that the differences between δ_SOM and δ_RHI are large enough to calculate the contributions of R_RHI, R_SOM and R_L to R_S based on the mass balance of the CO₂ isotope ratios. R_RHI and R_SOM had close relationships with soil temperature, and R_L was influenced by soil temperature and moisture. In summer, under high soil temperatures, R_RHI and R_SOM were the predominant sources of R_S and the proportion of R_RHI to R_SOM to R_L was 51:44:5. In winter, R_L was predominant and the proportion of R_RHI to R_SOM to R_L was 20:11:69. The estimated annual emissions of R_RHI, R_SOM and R_L were 1.45, 2.10 and 1.30 Mg C ha^?1, respectively; thus, the proportion of R_RHI to R_SOM to R_L was 30:43:27 on a whole-year basis.     

Elevated CO2 alters the structure of the bacterial community assimilating plant‐derived carbon in the rhizosphere of soya bean

Elevated CO₂ (eCO₂) increases rhizodeposits, which in turn alters the soil microbial community. However, it is not really known how the microbial community metabolizes plant‐derived carbon (C) in the rhizosphere under eCO₂, especially in agricultural soils. This study used a ¹³CO₂ labelling technique combined with DNA‐stable isotope probing (SIP) to fractionate the ¹³C‐DNA and ¹²C‐DNA from the rhizosphere of soya bean plants (Glycine max (L.) Merr. cv. Suinong 14) grown for 54 days under ambient CO₂ (aCO₂) (390 ppm) or eCO₂ (550 ppm). The DNA fractions were then subjected to Illumina Miseq sequencing. The results showed that eCO₂ decreased the richness and diversity of the ¹³C‐assimilating bacterial community compared to aCO₂ (p < 0.05). Elevated CO₂ decreased the abundances of genera, including Pseudarthrobacter, Gaiellales_uncultured, Microlunatus, Gemmatimonas, Gemmatimonadaceae_uncultured, Ramlibacter, Massilia, Luteimonas, Acidobacteriaceae_uncultured, Bryobacter and Candidatus_Solibacter. These genera were probably fast‐growing bacteria and sensitive to labile C. In contrast, eCO₂ stimulated the growth of genera Novosphingobium, Acidimicrobiales_uncultured, Bacillus, Flavisolibacter and Schlesneria, which were able to assimilate complex C compounds. Moreover, the increased population of Novosphingobium under eCO₂ might have accelerated electron flow from the oxidation of organic C. Correspondingly, eCO₂ did not affect the concentration of the dissolved organic C but increased the plant‐derived ¹³C in the rhizosphere. These results indicated that an eCO₂‐induced increase in non‐labile C in rhizodeposits contributed to the increase in population size of a number of the plant‐C‐metabolizing genera that might become the mechanism for the turnover of fresh C in the rhizosphere, modifying the soil C cycle under eCO₂ environments.     

用13C示踪研究CO2浓度倍增对枸杞光合产物积累的影响

CO2作为光合作用的底物,其浓度的高低直接影响植物光合作用能力的大小,为探究近年来大气CO2浓度升高对宁夏枸杞光合产物的影响,应用碳同位素示踪技术与开顶气室法,测定分析2种CO2浓度(倍增浓度(720±20)μmol/mol;对照(360±20)μmol/mol)处理下宁夏枸杞苗木各器官中光合产物积累与分配的差异。结果表明:CO2浓度倍增处理下,宁夏枸杞叶片净光合速率、胞间CO2浓度、气孔导度、水分利用效率均明显高于对照;在速生期,叶片蒸腾速率较对照显著降低,在生长后期没有差异。CO2浓度倍增处理下,未进行标记的宁夏枸杞各个器官的13C自然丰度值相对于对照均有不同程度的下降。13C标记90 d时,对宁夏枸杞各器官在24 h、48 h和7 d的δ13C值进行测定,并与未标记之前的δ13C值比较发现宁夏枸杞叶片δ13C值在24 h最大,根和茎δ13C值在标记结束后48 h最大,之后开始下降,根部δ13C值下降的幅度较小,光合产物总体呈现的是叶向茎,再向根部转移的规律。在CO2浓度倍增处理90和120 d时,宁夏枸杞δ13C值在枸杞根、茎和叶中均有不同程度的升高。处理90 d时δ13C值较对照增加的百分比分别为茎(65.53%)根(27.39%)叶(18.05%),120 d时为果实(145.04%)叶(143.56%)根(49.96%)茎(43.26%)。因此大气CO2浓度倍增,增强了宁夏枸杞的光合能力,增加了光合产物在各器官中的积累,在速生期光合产物在茎中的增加比例最大,而生长后期则在果实和叶中的增加比例较大。     

The effect of diet on isotopic turnover in Collembola examined using the stable carbon isotopic compositions of lipids

Combined compound-specific stable carbon isotopic methods and fatty acid abundance determinations have been used to examine feeding preferences and C allocation in organisms where direct observation of feeding is difficult. In order to examine the effect of differing diets on the δ¹³C values of fatty acids and sterols of Collembola, the diets of two collembolan species, Folsomia candida and Proisotoma minuta, were switched from a yeast diet to one of four isotopically distinct diets, and the δ¹³C values of the lipids monitored over the next 39 d. Cholesterol remained the only sterol detected in both collembolan species, despite the diets containing widely differing sterol compositions. The δ¹³C values of collembolan lipids recorded after long term feeding were often different to those of the same components in the diet, indicating that fractionation or partitioning occurs during digestion, assimilation and biosynthesis within the Collembola, thereby shifting consumer lipid δ¹³C values away from those of the corresponding dietary components. The rates of change of δ¹³C values differed among compounds, with half-lives ranging between 29 min and 14 d. Some of these differences appear to be related to the abundance of dietary components, such that fatty acids present in high abundance in the diet (e.g. 18:2(n−6)) were rapidly assimilated in high proportions into collembolan lipids, leading to a rapid change in δ¹³C values. Similarly, isotopic turnover in the 16:1(n−7) fatty acid, present in the newly presented diets in only low abundances, was significantly correlated to the rate of removal of this component from the consumer fatty acid pool. The rates of change of δ¹³C values in P. minuta lipids did not vary significantly with diet, whilst the rates of change of δ¹³C values of lipids in F. candida were affected by the diets the Collembola consumed. Results of an experiment providing F. candida and P. minuta with two diets of different quality demonstrated that F. candida responded to the high quality diet with increased growth and fecundity, whilst P. minuta responded with increased fecundity only. Thus, the abilities of the two species to respond to diets of varying quality, amongst other factors, is concluded to lead to differences in the rates of change of δ¹³C values reflecting differences in lipid turnover.     

Three-source partitioning of CO2 efflux from soil planted with maize by C natural abundance fails due to inactive microbial biomass

A theoretical approach to the partitioning of carbon dioxide (CO₂) efflux from soil with a C₃ vegetation history planted with maize (Zea mays), a C₄ plant, into three sources, root respiration (RR), rhizomicrobial respiration (RMR), and microbial soil organic matter (SOM) decomposition (SOMD), was examined. The δ¹³C values of SOM, roots, microbial biomass, and total CO₂ efflux were measured during a 40-day growing period. A three-source isotopic mass balance based on the measured δ¹³C values and on assumptions made in other studies showed that RR, RMR, and SOMD amounted to 91%, 4%, and 5%, respectively. Two assumptions were thoroughly examined in a sensitivity analysis: the absence of ¹³C fractionation and the conformity of δ¹³C of microbial CO₂ and that of microbial biomass. This approach strongly overestimated RR and underestimated RMR and microbial SOMD. CO₂ efflux from unplanted soil was enriched in ¹³C by 2.0‰ compared to microbial biomass. The consideration of this ¹³C fractionation in the mass balance equation changed the proportions of RR and RMR by only 4% and did not affect SOMD. A calculated δ¹³C value of microbial CO₂ by a mass balance equation including active and inactive parts of microbial biomass was used to adjust a hypothetical below-ground CO₂ partitioning to the measured and literature data. The active microbial biomass in the rhizosphere amounted to 37% to achieve an appropriate ratio between RR and RMR compared to measured data. Therefore, the three-source partitioning approach failed due to a low active portion of microbial biomass, which is the main microbial CO₂ source controlling the δ¹³C value of total microbial biomass. Since fumigation-extraction reflects total microbial biomass, its δ¹³C value was unsuitable to predict δ¹³C of released microbial CO₂ after a C₃-C₄ vegetation change. The second adjustment to the CO₂ partitioning results in the literature showed that at least 71% of the active microbial biomass utilizing maize rhizodeposits would be necessary to achieve that proportion between RR and RMR observed by other approaches based on ¹⁴C labelling. The method for partitioning total below-ground CO₂ efflux into three sources using a natural ¹³C labelling technique failed due to the small proportion of active microbial biomass in the rhizosphere. This small active fraction led to a discrepancy between δ¹³C values of microbial biomass and of microbially respired CO₂.     

Stable isotope analysis reveals whether soil-living elaterid larvae move between agricultural crops

Tracking the movement of soil-living herbivores is difficult, albeit important for understanding their spatial ecology as well as for pest management. In this study the movement of Agriotes obscurus larvae between plots harbouring isotopically different plants was examined. Neither between maize and wheat nor between maize and grassland movement could be detected. These data suggest that Agriotes larvae rarely disperse between crops as long as local food supply is sufficient. Moreover, the current approach provides a new means to study the dispersal of soil invertebrates in situ.     

Stable carbon isotope ratios of water-extractable organic carbon affected by application of rice straw and rice straw compost during a long-term rice experiment in Yamagata,Japan

ABSTRACT

Hot-water- and water-extractable organic matter were obtained from soil samples collected from a rice paddy 31 years after the start of a long-term rice experiment in Yamagata, Japan. Specifically, hot-water-extractable organic carbon and nitrogen (HWEOC and HWEON) were obtained by extraction at 80°C for 16 h, and water-extractable organic carbon and nitrogen (WEOC and WEON) were obtained by extraction at room temperature. The soil samples were collected from surface (0–15 cm) and subsurface (15–25 cm) layers of five plots that had been treated with inorganic fertilizers alone or with inorganic fertilizers plus organic matter, as follows: PK, NPK, NPK plus rice straw (RS), NPK plus rice straw compost (CM1), and NPK plus a high dose of rice straw compost (CM3). The soil/water ratio was 1:10 for both extraction temperatures. We found that the organic carbon and total nitrogen contents of the bulk soils were highly correlated with the extractable organic carbon and nitrogen contents regardless of extraction temperature, and the extractable organic carbon and nitrogen contents were higher in the plots that were treated with inorganic fertilizers plus organic matter than in the PK and NPK plots. The HWEOC and WEOC δ¹³C values ranged from ?28.2% to ?26.4% and were similar to the values for the applied rice straw and rice straw compost. There were no correlations between the HWEOC or WEOC δ¹³C values and the amounts of HWEOC or WEOC. The δ¹³C values of the bulk soils ranged from ?25.7% to ?23.2% and were lower for the RS and CM plots than for the PK and NPK plots. These results indicate that HWEOC and WEOC originated mainly from rice plants and the applied organic matter rather than from the indigenous soil organic matter. The significant positive correlations between the amounts of HWEOC and HWEON and the amount of available nitrogen (P < 0.001) imply that extractable organic matter can be used as an index for soil fertility in this long-term experiment. We concluded that the applied organic matter decomposed more rapidly than the indigenous soil organic matter and affected WEOC δ¹³C values and amounts.     

Trophic shift of stable isotopes and fatty acids in Collembola on bacterial diets

Fatty acid (FA) analysis is a promising tool to study trophic relationships in soil food webs. We determined FA biomarkers to trace bacterial food sources (Bacillus megaterium, Pseudomonas putida, Enterobacter aerogenes) of Collembola (Heteromurus nitidus, Protaphorura fimata, Folsomia candida). In addition, δ¹⁵N, δ¹³C, C/N ratio, body weight and NLFA/PLFA ratio (neutral lipid/phospholipid fatty acids) of Collembola were assessed. These measures indicated that P. putida ranked first, B. megaterium second and E. aerogenes third in food quality. FAs specific for bacteria were found in the NLFAs of the Collembola reflecting the respective bacterial diet. Biomarker FAs for gram-positive bacteria were methyl branched i14:0, i15:0, a15:0 and i17:0. Consumption of gram-negative bacteria was reflected by the cyclic form cy17:0 (E. aerogenes, P. putida) and by 16:1ω5 (P. putida).     

土壤呼吸及其13C同位素测定方法的对比研究

为比较不同方法在土壤呼吸及其_¹³C同位素测定中的差异,我们应用几种常见的方法测定了不同有机质含量的水稻土壤在一定时间内的CO₂排放量及_¹³C-CO₂丰度,以期准确评估土壤呼吸及碳排放,并为相关研究提供参考。本实验采用了气相色谱仪法(GC-TCD)、稳定同位素比值质谱仪气体进样法(Gasbench-IRMS)、甲酚红显色法(MicroResp)、碱液吸收法四种方法测定土壤呼吸速率;Gasbench-IRMS法和碱液吸收法两种方式检测土壤呼吸的_¹³CO₂含量。结果表明,(1)两种仪器法(GC、IRMS)测定土壤呼吸速率的数值结果相近(基础呼吸)或趋势一致(诱导呼吸),且重复性好（标准差分别为0.011、0.010 mg C /kg/h）,准确度高;MicroResp法的测定结果与仪器测量值较为相近,但分辨率较低;碱液吸收法的测定结果较真实值偏高(当土壤有机质含量低时)或偏低(当土壤有机质含量高时)。(2)在测定CO₂中的_¹³C含量上,Gasbench-IRMS直接测定的结果误差小(δ_¹³C值的标准偏差为0.137‰),接近实际值,可以准确地反应出土壤微生物呼吸时对底物的利用状况。综上,仪器法较化学分析法(MicroResp、碱液吸收)更能准确测定土壤呼吸及其_¹³C同位素。     

Response of microbial communities to water stress in irrigated and drought-prone tallgrass prairie soils

To better understand how water stress and availability affect the structure of microbial communities in soil, I measured the change in phospholipid fatty acids (PLFA) and the incorporation of ¹³C-labeled glucose into the PLFA following exposure to water stress. Overlaid on the laboratory water stress treatment, samples were collected from drought-prone and irrigated (11 years) tallgrass prairie soil (0-10 cm depth). In the laboratory, soils were either incubated at −250 kPa or dried steadily over a 3-d period to −45 MPa. On the fourth day, the dried samples were brought up to −250 kPa and then all samples received 250 μg of glucose-C (+4000 δ¹³C-PDB) solution that brought them to −33 kPa matric water potential. Samples were then extracted for PLFA following 6 and 24 h of incubation (25 °C). Non-metric multidimensional scaling (NMS) techniques and multi-response permutation procedure (MRPP) showed that the largest effect on the mol% distribution of PLFA was related to the field scale water addition experiment. In response to irrigation, the PLFA 16:1ω5, 18:1+, and 18:2ω6,9 showed increases, and a15:0, a17:0, and cy19:0 showed decreases in their respective mol%. Effects related to the induction of laboratory water stress were predominantly associated with a decrease in the mol% distribution of the putative fungal biomarker (18:2ω6,9) with little to no change in the mol% distribution of the bacterial biomarkers. Interestingly, the flow of C to the microbial community was not strongly related to any single PLFA, and differences were rather subtle, but multivariate MRPP detected change to the community structure related to the laboratory water stress treatment but not related to the 11 years of field irrigation. Our results suggest that both the total and the actively metabolizing bacterial community in soil were generally resistant to the effects of water stress brought by rewetting of dry soil. However, more research is needed to understand the nature of the fungal response to drying and rewetting in soil.