In situ simultaneous measuring method for the determination of key processes of soil organic carbon cycling: Soil microbial respiration using laser spectrometry.

RATIONALE: Soil microbial heterotrophic C-CO2 respiration is important for C cycling. Soil CO2 differentiation and quantification are vital for understanding soil C cycling and CO2 emission mitigation. Presently, soil microbial respiration (SR) quantification models are based on native soil organic matter (SOM) and require consistent monitoring of δ13C and CO2. METHODS: We present a new apparatus for achieving in situ soil static chamber incubation and simultaneous CO2 and δ13C monitoring by cavity ring-down spectroscopy (CRDS) coupled with a soil culture and gas introduction module (SCGIM) with multi-channel. After a meticulous five-point inter-calibration, the repeatability of CO2 and δ13C values by using CRDS-SCGIM were determined, and compared with those obtained using gas chromatography (GC) and isotope ratio mass spectrometry (IRMS), respectively. We examined the method regarding quantifying SR with various concentrations and enrichment of glucose and then applied it to investigate the responses of SR to the addition of different exogenous organic materials (glucose and rice residues) into paddy soils during a 21-day incubation. RESULTS: The CRDS-SCGIM CO2 and δ13C measurements were conducted with high precision (< 1.0 µmol/mol and 1‰, respectively). The optimal sampling interval and the amount added were not exceeded 4 h and 200 mg C/100 g dry soil in a 1 L incubation bottle, respectively; the 13C-enrichment of 3%-7% was appropriate. The total SR rates observed were 0.6-4.2 µL/h/g and the exogenous organic materials induced -49%-28% of priming effects in native SOM mineralisation. CONCLUSIONS: Our results show that CRDS-SCGIM is a method suitable for the quantification of soil microbial CO2 respiration, requiring less extensive lab resources than GC/IRMS.

Rapid, sensitive analysis method for determining the nitrogen stable isotope ratio of total free amino acids in soil.

RATIONALE: The amino acid-nitrogen (AA-N) isotope analysis of naturally abundant or isotope-labeled samples is indispensable for tracing nitrogen transfer in soil nitrogen biogeochemical cycling processes. Despite the usefulness of AA-N isotope analysis, the preparation methods are complex and time-consuming, and necessitate the use of toxic reagents. METHODS: We present an improved, rapid method for AA-N isotope analysis with high precision. At a high pH, AA-N was released and oxidized to N2 O using ClO- under vacuum. Additionally, purge-and-trap isotope ratio mass spectrometry was used to analyze N2 O. Moreover, we investigated the effect of various factors on the N2 O conversion process with glycine and applied the results to seven representative single-N AAs (alanine, serine, cysteine, aspartic acid, glutamic acid, leucine, and phenylalanine) and five poly-N AAs (lysine, arginine, histidine, tryptophan, and asparagine), as well as side-chain analogs, blank reagent, and other N forms. RESULTS: The concentration of ClO- and the pH were determined to be crucial factors for achieving desirable AA-N to N2 O conversion efficiencies. Glycine-N had the highest N2 O yield of 70%, with isotopic results consistent with those of the reference values at a high precision (within 0.5‰ for natural abundance and 0.01 atom% for 15 N-enrichment) at the nanomolar N level. Additionally, the α-NH2 AAs were labile, and the single-N AAs were more easily converted to N2 O than poly-N AAs. With the exception of γ-aminobutyric acid, the N2 O conversion efficiencies of the side-chain N analogs were very low (below 5%). This method was also applicable to the 15 N analysis of the total free AAs in complex soil samples without interference from analytical blanks and other forms of N. CONCLUSIONS: Our method is highly selective for the α-NH2 groups of an amino acid, and the oxidation of the side chain is difficult. In addition, the method is sensitive, rapid, and convenient, and does not require toxic reagents.

Visualization and quantification of carbon "rusty sink" by rice root iron plaque: Mechanisms, functions, and global implications.

Paddies contain 78% higher organic carbon (C) stocks than adjacent upland soils, and iron (Fe) plaque formation on rice roots is one of the mechanisms that traps C. The process sequence, extent and global relevance of this C stabilization mechanism under oxic/anoxic conditions remains unclear. We quantified and localized the contribution of Fe plaque to organic matter stabilization in a microoxic area (rice rhizosphere) and evaluated roles of this C trap for global C sequestration in paddy soils. Visualization and localization of pH by imaging with planar optodes, enzyme activities by zymography, and root exudation by 14 C imaging, as well as upscale modeling enabled linkage of three groups of rhizosphere processes that are responsible for C stabilization from the micro- (root) to the macro- (ecosystem) levels. The 14 C activity in soil (reflecting stabilization of rhizodeposits) with Fe2+ addition was 1.4-1.5 times higher than that in the control and phosphate addition soils. Perfect co-localization of the hotspots of ß-glucosidase activity (by zymography) with root exudation (14 C) showed that labile C and high enzyme activities were localized within Fe plaques. Fe2+ addition to soil and its microbial oxidation to Fe3+ by radial oxygen release from rice roots increased Fe plaque (Fe3+ ) formation by 1.7-2.5 times. The C amounts trapped by Fe plaque increased by 1.1 times after Fe2+ addition. Therefore, Fe plaque formed from amorphous and complex Fe (oxyhydr)oxides on the root surface act as a "rusty sink" for organic matter. Considering the area of coverage of paddy soils globally, upscaling by model revealed the radial oxygen loss from roots and bacterial Fe oxidation may trap up to 130 Mg C in Fe plaques per rice season. This represents an important annual surplus of new and stable C to the existing C pool under long-term rice cropping.

[Response of Extracellular Enzyme Activities to Substrate Availability in Paddy Soil with Long-term Fertilizer Management].

The availability of carbon (C), nitrogen (N), and other substrates in soil determines the growth and metabolism of microorganisms and affects the activity of extracellular enzymes. To study the activities of ß-1,4-glucosidase (BG) and ß-1,4-N-acetylglucosaminidase (NAG) in response to C and N availability, samples that underwent four treatments-non-fertilization (CK), chemical fertilizer (NPK), combination of organic manure and chemical fertilizer (OM), and mixture of straw and chemical fertilizer (ST)-were collected from long-term fertilization paddy soil and incubated for 0, 4, 8, and 12 months to obtain soil with different C and N availability gradients. The results showed that the dissolved organic carbon(DOC) content of OM and ST treatment samples was 2-3 times higher than that of CK and NPK treatment samples. With the increase of DOC and ammonium (NH4+-N) contents, the activities of BG and NAG and the contents of microbial biomass C (MBC) and N (MBN) showed no increase during incubation within each treatment. Fertilization treatments, incubation time, and their interaction are crucial factors varying the contents of DOC, NH4+-N, MBC, and MBN among different fertilization treatments (P<0.01). There was a positive correlation between MBC/MBN and DOC/NH4+-N of OM treatment (P<0.05) and a negative relationship between ln(BG)/ln(NAG) and DOC/NH4+-N of ST treatment (P<0.01), indicating that the availability of substrates played a key role in the potential activity of extracellular enzymes in paddy soil, and the carbon-nitrogen ratio of microbial biomass was controlled by the C/N stoichiometry of substrates in soil. The results have a certain guiding significance for further study on the variation of extracellular enzyme activity in paddy soil, regulating the balance of carbon and nitrogen, and improving the fertility of paddy soil.

[Allocation and Stabilization Responses of Rice Photosynthetic Carbon in the Plant-Soil System to Phosphorus Application].

This research studied the response of the input and allocation of photosynthetic carbon (C) to phosphorus (P) in paddy soils. Two treatments were conducted in this experiment:no P application (P0) and the application of 80 mg·kg-1 of P (P80). The rice cultivar was the indica Zhongzao 39. The 13C-CO2 continuous labeling technique was used to identify the photosynthetic C distribution of the rice. The results showed that the application of P80 significantly increased the photosynthates allocation in the rice aboveground, but reduced their allocation in the rhizosphere soil (P<0.05). At the jointing stage, P80 application increased the photosynthetic C content of the rice by 70%, but the root dry weight decreased 31%. Compared with P0, the total C content of the aboveground rice was increased 0.31 g·pot-1 by P80. The ratio of rice roots to shoots decreased with the P80 treatment. Moreover, P80 application led to an increase in the photosynthetic microbial biomass in the non-rhizosphere soil C (13C-MBC) of 0.03 mg·kg-1, but still decreased its allocation in the rhizosphere soil. The allocation of photosynthetic C to the particulate organic matter fraction (POC) and mineral fraction (MOC) in the non-rhizosphere soil showed no significant differences between P0 and P80. Additionally, the P80 fertilization treatment significantly lowered the content of POC in the rhizosphere soil. In summary, P application increased the allocation of photosynthetic C in the soil-rice system, but reduced the accumulation of photosynthetic C in the soil. This research provided a theoretical basis and data supporting the rational application of P fertilizer, and was also of great significance as a study of the transportation and allocation of photosynthetic C and its sequestration potential response to the application of P to the rice soil.

[Characterization of Soil Organic Carbon Mineralization Under Different Gradient Carbon Loading in Paddy Soil].

Available carbon is the most active part of the soil carbon pool. It is also the main carbon source of soil microbes and plays an important role in the processes of soil organic carbon mineralization and accumulation. However, the mechanisms are still not clear how soil organic carbon mineralization and its priming effect (PE) are affected by different input levels of readily available carbon, based on the growth requirements of microbes in paddy soil. In this study, an incubation experiment was conducted by adding different levels (0.5, 1, 3, and 5 times of MBC) of exogenous source organic carbon (13C-glucose) to the soil. The mineralization dynamics of labile organic carbon and its priming effect was investigated. The mineralization rate of glucose-C increased significantly with the increasing carbon loading level. The distribution of glucose-C into rapid and slow C pools was also exponentially correlated with the carbon loading (R2=0.99, P<0.05 and R2=0.99, P<0.05, respectively). Negative PE was observed at high carbon loading (3×MBC and 5×MBC); while positive PE was induced by low carbon loading (0.5×MBC and 1×MBC). The cumulative PE was 160.0 mg·kg-1 and 325.1 mg·kg-1, respectively, at the end of the incubation. Redundancy analysis showed that the main factors affecting the cumulative PE were MBC, MBN, and DOC at the initial glucose mineralization stage, while ß-glucosidase, chitinase, and ammonium nitrogen were the main factors at later stages. Therefore, the readily available carbon loading has an important effect on the organic carbon mineralization and PE in paddy soil. Higher carbon loading was good for the accumulation of organic carbon sequestration in paddy soil. This study is of great scientific significance for revealing the activity of organic carbon in paddy fields and for its contribution to the development of sustainable agriculture.

[Dynamics of Rice Photosynthesized Carbon Input and Its Response to Nitrogen Fertilization at the Jointing Stage:13 C-CO2 Pulse-labeling].

Photosynthesized carbon (C) is an important source of soil organic C in paddy fields, and its input and distribution are affected by rice growth and soil fertility. Fertilizer application plays an important role in rice growth. The 13C pulse-labeling method was used to quantify the dynamics and distribution of input photosynthesized C in the rice-(rhizosphere-and bulk-) soil system and its response to nitrogen fertilizer (N) application. The results suggested that N fertilization significantly increased the rice aboveground and the root biomass and decreased the rice biomass root/shoot ratio. The amount of assimilated 13C gradually decreased in the rice plants but gradually decreased over 0-6 days and increased over 6-26 days in the rhizosphere and bulk soil during rice growth. N fertilization significantly increased the amount of assimilated 13C in the rhizosphere soil by 9.5%-32.6% compared with the control. In comparison to the unfertilized treatment, the application of N fertilization resulted in higher photosynthetic13C in rice aboveground and in the root by 24.5%-134.7% and 9.1%-106%, respectively. With the N fertilized and unfertilized treatments, 85.5%-93.2% and 91.3%-95.7%, respectively, of input photosynthetic 13C was distributed in the rice plants. The results suggested that N fertilization significantly affected the distribution of photosynthesized C in the rice-soil system (P<0.01). After 26 days of pulse labeling, the distribution of photosynthetic 13C into rice aboveground was increased by 13.4%, while the distribution into the rhizosphere and bulk soil were decreased by 21.9% and 52.2%, respectively, in the N fertilized treatments compared with the unfertilized treatments. Therefore, the N application increased the distribution of photosynthesized carbon in the soil-rice system but decreased the accumulation in the rhizosphere and bulk soil. The findings of this study provided a theoretical basis for our understanding of the dynamic of photosynthetic C in the plant-soil system and the assimilation of the soil organic matter pool in the paddy soil ecosystem.

[Effect of CO2 Doubling and Different Plant Growth Stages on Rice Carbon, Nitrogen, and Phosphorus and Their Stoichiometric Ratios].

The variation characteristics of ecological stoichiometric ratios can reflect the nature of plant adaptation to environmental changes. The C, N, and P contetns, and their stoichiometric ratios in different organs of rice were studied using a CO2 continuous labeling system, by simulating the increase of atmospheric CO2 concentration (800×10-6). The results showed that CO2 doubling promoted the growth of rice organs and increased the root/shoot ratio. CO2 doubling reduced the shoot TN content in different growth periods, increased the C/N ratio in the rice root, shoot, and grain, decreased the N use efficiency, and improved the P use efficiency. Multiple comparison and Venn diagram analyses showed that CO2 concentration only has a significant impact on the TN content in the rice shoot; it contributed little to the variation in rice nutrient content and their stoichiometric ratios, indicating that CO2 doubling had no effect on these. Under the condition of elevated atmospheric CO2 concentrations, the C, N, and P contents and their stoichiometirc ratios, in rice organs had good homeostasis, and the stoichiometric change during growth periods was consistent with "the Growth Rate Theory". In farmland management, appropriate nitrogen fertilizers can alleviate the nutrient balance pressure caused by the increase in CO2 concentration.

Influence of land use on bacterial and archaeal diversity and community structures in three natural ecosystems and one agricultural soil.

Studying shifts in microbial communities under different land use can help in determining the impact of land use on microbial diversity. In this study, we analyzed four different land-use types to determine their bacterial and archaeal diversity and abundance. Three natural ecosystems, that is, wetland (WL), grassland (GL), and forest (FR) soils, and one agricultural soil, that is, tea plantation (TP) soil, were investigated to determine how land use shapes bacterial and archaeal diversity. For this purpose, molecular analyses, such as quantitative polymerase chain reaction (Q-PCR), 16S rRNA gene sequencing, and terminal restriction fragment length polymorphism (T-RFLP), were used. Soil physicochemical properties were determined, and statistical analyses were performed to identify the key factors affecting microbial diversity in these soils. Phylogenetic affiliations determined using the Ribosomal Database Project (RDP) database and T-RFLP revealed that the soils had differing bacterial diversity. WL soil was rich in only Proteobacteria, whereas GR soil was rich in Proteobacteria, followed by Actinobacteria. FR soil had higher abundance of Chloroflexi species than these soils. TP soil was rich in Actinobacteria, followed by Chloroflexi, Acidobacteria, Proteobacteria, and Firmicutes. The archaeal diversity of GL and FR soils was similar in that most of their sequences were closely related to Nitrososphaerales (Thaumarchaeota phylum). In contrast, WL soil, followed by TP soil, had greater archaeal diversity than other soils. Eight different archaeal classes were found in WL soil, and Pacearchaeota class was the richest one. The abundance of bacterial and archaeal 16S rRNA gene copies in WL and GL soils was significantly higher than that in FR and TP soils. Redundancy analysis showed that bacterial diversity was influenced by abiotic factors, e.g., total organic carbon and pH, whereas total nitrogen, pH, and cation exchange capacity (CEC) significantly affected archaeal community composition. Pearson correlation analysis showed that bacterial and archaeal 16S rRNA gene abundance had the highest correlation with clay content (r > 0.905, P < 0.01), followed by total-P, CEC, pH, and silt (%). These results will lead to more comprehensive understanding of how land use affects microbial distribution.

[Characteristic of Abundances and Diversity of Carbon Dioxide Fixation Microbes in Paddy Soils].

To get a better understanding of the microbial autotrophic carbon sequestration potential of paddy fields and its mechanisms, soil incubation experiment was conducted for four representative paddy soils. The molecular biological methods[quantitative PCR (qPCR), clone library and terminal-restriction fragment length polymorphism (T-RFLP) technique] based on cbbL and cbbM genes encoding the key enzymes[ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO)] of Calvin cycle were used to determine the abundance and diversity of autotrophic microbes. The results showed that, after 45 days of incubation, carbon dioxide fixation autotrophic microbial abundances were generally increased compared with those before incubation, and cbbL gene abundances were approximately three magnitudes higher than those of cbbM. Dominant microbial populations varied among the four paddy soils, and most of these OTUs were distantly related to known sequences, only part of them could be grouped into Proteobacteria and Actinobacteria. RDA analysis results showed that soil organic carbon (SOC), cation exchange capacity (CEC), pH, clay, silk and sand content had significant effects on the CO2 fixation microbial community. Consequently, the results of this study provide significant reference to understand the role of microorganisms in carbon cycle process. The results are helpful for providing a scientific basis for scientific management of paddy soil fertility and low carbon agriculture construction.

[Characteristics and Influencing Factors of Biologically-based Phosphorus Fractions in the Farmland Soil].

A suitable fractionation method of phosphorus (P) is a key to effective assessment of soil P componential features. Here a new biologically-based P (BBP) method was used to evaluate the P fractions in the upland and paddy soils across large-scale area in China. The soil P was divided into four components:① soluble or rhizosphere-intercepted (CaCl2-P), ② organic acid activated and inorganic weakly bound (Citrate-P), ③ enzyme mineralization of organic P (Enzyme-P), ④ potential activation of inorganic P (HCl-P). Then, the relationships between biologically-based P fractions and standard Olsen-P were investigated, and driving factors of P fractions were identified. The results showed that P content was in order of HCl-P>Citrate-P>Enzyme-P>CaCl2-P. All P components of upland soil displayed higher levels than those of paddy soil. Moreover, the P components were highly positively correlated with the Olsen-P, suggesting that each P component contributed to soil P availability. However, it was found that Olsen-P was most highly correlated with CaCl2-P and Enzyme-P (R2=0.359; R2=0.386) in upland soil, while Olsen-P was most highly with Citrate-P (R2=0.788) in paddy soil. This result indicated that available P of upland soil was mainly from organic P mineralization and soluble P, and available P in paddy soil was mainly from inorganic P activation. Redundancy analysis (RDA) showed that the P components were mainly affected by soil pH and silt content, which suggested that it could enhance the P availability via regulating soil pH in the agricultural activities.

Soil Carbon-Fixation Rates and Associated Bacterial Diversity and Abundance in Three Natural Ecosystems.

CO2 assimilation by autotrophic microbes is an important process in soil carbon cycling, and our understanding of the community composition of autotrophs in natural soils and their role in carbon sequestration of these soils is still limited. Here, we investigated the autotrophic C incorporation in soils from three natural ecosystems, i.e., wetland (WL), grassland (GR), and forest (FO) based on the incorporation of labeled C into the microbial biomass. Microbial assimilation of 14C (14C-MBC) differed among the soils from three ecosystems, accounting for 14.2-20.2% of 14C-labeled soil organic carbon (14C-SOC). We observed a positive correlation between the cbbL (ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) large-subunit gene) abundance, 14C-SOC level, and 14C-MBC concentration confirming the role of autotrophic bacteria in soil carbon sequestration. Distinct cbbL-bearing bacterial communities were present in each soil type; form IA and form IC RubisCO-bearing bacteria were most abundant in WL, followed by GR soils, with sequences from FO soils exclusively derived from the form IC clade. Phylogenetically, the diversity of CO2-fixing autotrophs and CO oxidizers differed significantly with soil type, whereas cbbL-bearing bacterial communities were similar when assessed using coxL. We demonstrate that local edaphic factors such as pH and salinity affect the C-fixation rate as well as cbbL and coxL gene abundance and diversity. Such insights into the effect of soil type on the autotrophic bacterial capacity and subsequent carbon cycling of natural ecosystems will provide information to enhance the sustainable management of these important natural ecosystems.

Effect of simulated tillage on microbial autotrophic CO2 fixation in paddy and upland soils.

Tillage is a common agricultural practice affecting soil structure and biogeochemistry. To evaluate how tillage affects soil microbial CO2 fixation, we incubated and continuously labelled samples from two paddy soils and two upland soils subjected to simulated conventional tillage (CT) and no-tillage (NT) treatments. Results showed that CO2 fixation ((14)C-SOC) in CT soils was significantly higher than in NT soils. We also observed a significant, soil type- and depth-dependent effect of tillage on the incorporation rates of labelled C to the labile carbon pool. Concentrations of labelled C in the carbon pool significantly decreased with soil depth, irrespective of tillage. Additionally, quantitative PCR assays revealed that for most soils, total bacteria and cbbL-carrying bacteria were less abundant in CT versus NT treatments, and tended to decrease in abundance with increasing depth. However, specific CO2 fixation activity was significantly higher in CT than in NT soils, suggesting that the abundance of cbbL-containing bacteria may not always reflect their functional activity. This study highlights the positive effect of tillage on soil microbial CO2 fixation, and the results can be readily applied to the development of sustainable agricultural management.

[Effects of Soil Texture on Autotrophic CO2 Fixation Bacterial Communities and Their CO2 Assimilation Contents].

Autotrophic bacteria can assimilate atmospheric carbon dioxide (CO2) and convert CO2 into organic carbon. The CO2 fixation by autotrophic bacteria is important for the improvement of carbon sequestration in agricultural soils. However, the effect of soil texture on autotrophic CO2 fixation bacteria and their CO2 fixation capacity is still unknown. Here, two paddy soils with different textures (loamy clay soil and sand clay loam soil) were incubated with continuous 14C-CO2 in a glass chamber. The two soils were developed from the same parent. At the end of 110 days incubation, the 14C-CO2 incorporated in soil organic carbon (14C-SOC), microbial biomass carbon (14C-MBC) and dissolved organic carbon (14C-DOC) were measured to explore the effects of soil texture on the autotrophic bacterial CO2 fixation rates. The effect of soil texture on the composition and diversity of autotrophic CO2 fixation bacterial community was investigated using cloning and sequencing of the cbbL gene, which encodes ribulose-1,5-biphosphate carboxylase/oxygenase (RubisCO) in the Calvin cycle. The results showed that the average contents of 14C-SOC, 14C-MBC and 14C-DOC were 133.81, 40.16 and 8.10 mg·kg-1 in loamy clay soil, respectively, which were significantly higher than their corresponding contents in sand clay loam soil (P<0.05). This suggested that soil texture not only affected the amounts of autotrophic bacteria CO2 fixation but also had an effect on the transformation of microbial assimilated 14C in soil. The cbbL gene libraries of two soils were significantly different as revealed by libshuff analyses (P<0.05). Phylogenetic analysis showed that cbbL sequences from the loamy clay soil were closely affiliated with known cultures such as Rhodoblastus acidophilus, Blastochloris viridis, Thauera humireducens, Mehylibium sp.and Variovorax sp., whereas these sequences belonging to the sand clay loam soil were related to branching lineages originating from Rhizobiales and Actinomycetales.Rarefaction curve, clone library coverage and diversity index analysis based on bacterial cbbL clone libraries indicated that the loamy clay soil had higher cbbL gene diversity compared to the sand clay loam soil. These results suggested that soil texture had a pronounced effect on the composition and diversity of autotrophic CO2 fixation bacterial communities. The higher clay content, nutrient availability and cation exchange capacity may stimulate the growth and activity of autotrophic bacteria, and result in the higher amounts of 14C in loamy clay soil. These data broaden the understanding and knowledge of mechanisms of microbial carbon fixation and their influencing factors in agricultural soils.

[Effects of long-term fertilization on bacterial and archaeal diversity and community structure within subtropical red paddy soils].

Paddy soils not only function as an important sink for "missing carbon" but also play an important role in the production of greenhouse gases such as N2O and CH4. Dynamic changes in greenhouse gases in the atmosphere are closely related to microbially mediated carbon and nitrogen transformation processes occurring in soil. Using soil samples collected from a long-term fertilization experimental site in Taojiang County, subtropical China (established in 1986), we determined the effects of long-term (>25 years) non-fertilization (CK), chemical fertilization (NPK), and NPK combined with rice straw residues (NPKS) on soil bacterial and archaeal community structures. The 16S rRNA genotypes from the three differently treated soils were divided into 9 bacterial phylotypes, mainly including Proteobacteria, Acidobacteria, Chloroflexi, and archaea of Crenarchaeota and Euryarchaeota. The relative abundance of Proteobacteria, Acidobacteria and Crenarchaeota increased in the soils under NPK and NPKS treatments, with the increase being greater in the latter treatment. LUBSHUFF statistical analyses also demonstrated that there was significant difference among the microbial community compositions in CK-, NPK- and NPKS-treated soils. The abundance of bacterial and archaeal 16S rRNA genes ranged from 0.58 x 10(10) to 1.06 x 10(10) copies · g(-1) dry soil and from 1.16 x 10(6) to 1.72 x 10(6) copies · g(-1) dry soil, respectively. Application of fertilizers increased the bacterial and archaeal abundance and diversity in the treated soils, with NPKS > NPK. Long-term chemical and organic applications significantly affected the abundance, diversity and composition of bacterial and archaeal communities in paddy ecosystems.

Cropping systems modulate the rate and magnitude of soil microbial autotrophic CO2 fixation in soil.

The effect of different cropping systems on CO2 fixation by soil microorganisms was studied by comparing soils from three exemplary cropping systems after 10 years of agricultural practice. Studied cropping systems included: continuous cropping of paddy rice (rice-rice), rotation of paddy rice and rapeseed (rice-rapeseed), and rotated cropping of rapeseed and corn (rapeseed-corn). Soils from different cropping systems were incubated with continuous (14)C-CO2 labeling for 110 days. The CO2-fixing bacterial communities were investigated by analyzing the cbbL gene encoding ribulose-1,5-bisphosphate carboxylase oxygenase (RubisCO). Abundance, diversity and activity of cbbL-carrying bacteria were analyzed by quantitative PCR, cbbL clone libraries and enzyme assays. After 110 days incubation, substantial amounts of (14)C-CO2 were incorporated into soil organic carbon ((14)C-SOC) and microbial biomass carbon ((14)C-MBC). Rice-rice rotated soil showed stronger incorporation rates when looking at (14)C-SOC and (14)C-MBC contents. These differences in incorporation rates were also reflected by determined RubisCO activities. (14)C-MBC, cbbL gene abundances and RubisCO activity were found to correlate significantly with (14)C-SOC, indicating cbbL-carrying bacteria to be key players for CO2 fixation in these soils. The analysis of clone libraries revealed distinct cbbL-carrying bacterial communities for the individual soils analyzed. Most of the identified operational taxonomic units (OTU) were related to Nitrobacter hamburgensis, Methylibium petroleiphilum, Rhodoblastus acidophilus, Bradyrhizobium, Cupriavidus metallidurans, Rubrivivax, Burkholderia, Stappia, and Thiobacillus thiophilus. OTUs related to Rubrivivax gelatinosus were specific for rice-rice soil. OTUs linked to Methylibium petroleiphilum were exclusively found in rice-rapeseed soil. Observed differences could be linked to differences in soil parameters such as SOC. We conclude that the long-term application of cropping systems alters underlying soil parameters, which in turn selects for distinct autotrophic communities.

Abundance and Diversity of CO2-Assimilating Bacteria and Algae Within Red Agricultural Soils Are Modulated by Changing Management Practice.

Elucidating the biodiversity of CO(2)-assimilating bacterial and algal communities in soils is important for obtaining a mechanistic view of terrestrial carbon sinks operating at global scales. "Red" acidic soils (Orthic Acrisols) cover large geographic areas and are subject to a range of management practices, which may alter the balance between carbon dioxide production and assimilation through changes in microbial CO(2)-assimilating populations. Here, we determined the abundance and diversity of CO(2)-assimilating bacteria and algae in acidic soils using quantitative PCR and terminal restriction fragment length polymorphism (T-RFLP) of the cbbL gene, which encodes the key CO(2) assimilation enzyme (ribulose-1,5-bisphosphate carboxylase/oxygenase) in the Calvin cycle. Within the framework of a long-term experiment (Taoyuan Agro-ecosystem, subtropical China), paddy rice fields were converted in 1995 to four alternative land management regimes: natural forest (NF), paddy rice (PR), maize crops (CL), and tea plantations (TP). In 2012 (17 years after land use transformation), we collected and analyzed the soils from fields under the original and converted land management regimes. Our results indicated that fields under the PR soil management system harbored the greatest abundance of cbbL copies (4.33 × 10(8) copies g(-1) soil). More than a decade after converting PR soils to natural, rotation, and perennial management systems, a decline in both the diversity and abundance of cbbL-harboring bacteria and algae was recorded. The lowest abundance of bacteria (0.98 × 10(8) copies g(-1) soil) and algae (0.23 × 10(6) copies g(-1) soil) was observed for TP soils. When converting PR soil management to alternative management systems (i.e., NF, CL, and TP), soil edaphic factors (soil organic carbon and total nitrogen content) were the major determinants of bacterial autotrophic cbbL gene diversity. In contrast, soil phosphorus concentration was the major regulator of algal cbbL community composition. Our results provide new insights into the diversity, abundance, and modulation of organisms responsible for microbial autotrophic CO(2) fixation in red acidic soils subjected to changing management regimes.

[Effects of Different Plantation Type on the Abundance and Diversity of Soil Microbes in Subtropical Red Soils].

Soil microbe plays an important role in carbon cycling, however, the effect of land use on soil microbe remain unclear. In present study, soil samples were collected from a long-term field experiment (Pantang Agroecosystem) in subtropical China (established in 1989), including paddy-rice (PR), upland-crop (UC), and paddy rice-upland crop rotation (PU) on soil bacterial (bacteria and Archaea) community structures. The effects of long-term different land uses were determined using terminal restriction fragment length polymorphism (T-RFLP) and quantitative PCR (RT-PCR) of the 16S rRNA gene. The abundance of soil microbial 16S rRNA genes ranged from 2.5 x 10(9)-1.5 x 10(10) copies x g(-1) dry soil. Compared with the PR, UP and UC led to a significant reduction in 16S rRNA genes abundance (P < 0.05). The soil microbial communities were dominated by bacteria such as Proteobacteria (76 and 90 and 327 bp; relative abundance of 47% - 53%) and Chloroflexi (65 bp; relative abundance of 10% - 12%). RDA statistical analyses demonstrated that there were significant differences in the microbial community composition in PR, UC, and PU treated soils. Soil organic carbon and total nitrogen content were the most highly statistically significant factors which positively influenced the soil microbial population. Taken together, our findings prove the long-term different land uses significantly influence the microbial diversity and community structure. The rice planting is an effective way of sustainable utilization of subtropical red soil, and it is more advantageous to the accumulation of soil organic matter, soil fertility and microbial diversity.

[Characteristics of the mineralization and transformation of autotrophic microbes-assimilated carbon in upland and paddy soils].

In this study, the mineralization and decomposition of autotrophic microbe assimilated carbon (new carbon) and native organic carbon in three upland and three paddy soils in subtropical China were measured using the 14C-labelled tracer technique. The results showed that, during the 100-d incubation, the mineralization of the 'new carbon' displayed three stages: a rise in the first 10 days, a slowdown from 11-d to 50-d, and a stabilization stage after 50 d. The mineralization ratio of the 'new carbon' ranged between 8.0% and 26.9% and the mineralization rate ranged from 0.01 to 0.22 microg 14C x g(-1) x d(-1) (0.01-0.22 microg 14C x g(-1) x d(-1) in paddy soils and 0.01-0.08 microg 14C x g(-1) x d(-1) in upland soils). However, the mineralization ratio and rate for native SOC were 1.55%-5.74% and 1.3-25.66 microg C x g(-1) x d(-1), respectively. In the soil active C pools, the 14C-dissolved organic carbon (DOC) first rose by as much as 0.3 mg x kg(-1) in the early stages of incubation (0-10 d), decreased rapidly by 0.42 mg x kg(-1) from 10-30 d, and then declined gradually. The fluctuation of the 14C-microbial biomass carbon (MBC) differed from that of the 14C DOC. At the beginning stage of the incubation (0-10 d), the 14C-MBC decreased rapidly, and then rapidly increased from 10 to 30 d, and the rate of increase reduced and was gradually stabilized after 40 d. The 14C-DOC/DOC renewal rate in the paddy soil was significantly higher than in the upland soil while the 14C-MBC/MBC renewal rate in the upland soil was significantly greater than in the paddy soil.

[Input and distribution of rice photosynthesized carbon in the tillering stage under different nitrogen application following continuous 13C labeling].

The input of rice-photosynthesized carbon (C) into soil plays an important role in soil C cycling. A 13C-labelled microcosm experiment was carried out to quantify the input of photosynthesized C into soil C pools in a rice-soil system during the tillering stage. Growing rice (Oryza sativa L. ) was continuously fed with 13C-labeled CO, ( C-CO, ) in a closed chamber without nitrogen (NO), or at different rates of N supply (N10,N20, N30, N40 or N60). The results showed that there were significant differences in rice shoot (1.58 g plot-1 to 4.35 g plot-1) and root (1.05 g plot-1 to 2.44 g plot-1 ) biomass among the N treatments after labeling for 18 days. The amounts of 13C in shoots and roots ranged from 44.0 g plot-1 to 157.6 g.plot-1 and 8.3 g.plot-1 to 49.4 g.plot-1, respectively, and generally followed the order of N60 > N40 > N20 > N10 > NO. The contents of rice-planted 13C-SOC, 13C-DOC and 13C-MBC in soil carbon pool were much higher than those of CK (without rice and N supply). The amount of 13C-SOC ranged from 11.1 g plot - to 23.7 gplot-1 , depending on the rate of N addition, accounting for 10.2% -18. 1% of the net assimilation. The amounts of 13C-DOC and 13C-MBC ranged from 4. 82-14.51 microg kg-1 and 526. 1-1 478.8 microg kg-1 , both depending on the N application rate. In addition, at 18-day of labeling, the 13C-SOC, 13C-DOC and 13C-MBC concentration was positively correlated with the rice biomass. Therefore, our results suggest that paddy soils can probably sequester more C from the atmosphere if more photosynthesized C enters the soils and N application can stimulate C rhizodeposition during the tillering stage.