Microbial iron reduction compensates for phosphorus limitation in paddy soils.

Limitation of rice growth by low phosphorus (P) availability is a widespread problem in tropical and subtropical soils because of the high content of iron (Fe) (oxyhydr)oxides. Ferric iron-bound P (Fe(III)-P) can serve as a P source in paddies after Fe(III) reduction to Fe(II) and corresponding H2PO4- release. However, the relevance of reductive dissolution of Fe(III)-P for plant and microbial P uptake is still an open question. To quantify this, 32P-labeled ferrihydrite (30.8 mg P kg-1) was added to paddy soil mesocosms with rice to trace the P uptake by microorganisms and plants after Fe(III) reduction. Nearly 2% of 32P was recovered in rice plants, contributing 12% of the total P content in rice shoots and roots after 33 days. In contrast, 32P recovery in microbial biomass decreased from 0.5% to 0.08% of 32P between 10 and 33 days after rice transplantation. Microbial biomass carbon (MBC) and dissolved organic C content decreased from day 10 to 33 by 8-54% and 68-77%, respectively, suggesting that the microbial-mediated Fe(III) reduction was C-limited. The much faster decrease of MBC in rooted (by 54%) vs. bulk soil (8-36%) reflects very fast microbial turnover in the rice rhizosphere (high C and oxygen inputs) resulting in the mineralization of the microbial necromass. In conclusion, Fe(III)-P can serve as small but a relevant P source for rice production and could partly compensate plant P demand. Therefore, the P fertilization strategies should consider the P mobilization from Fe (oxyhydr)oxides in flooded paddy soils during rice growth. An increase in C availability for microorganisms in the rhizosphere intensifies P mobilization, which is especially critical at early stages of rice growth.

Flooding and straw returning regulates the partitioning of soil phosphorus fractions and phoD-harboring bacterial community in paddy soils.

Flooding and straw returning are effective agricultural practices in promoting phosphorus (P) availability in paddy soils. However, little is known about the effects of these practices and their interaction on the soil P pools and functional microbes responsible for soil P mobilization. Our 4-year paddy field experiment aimed to analyze the responses of soil P fractions and phoD-harboring bacterial communities in a double-rice cropping system to intermittent flooding (IF) and continuous flooding (CF), in plots with (+ S) and without (-S) straw return. Compared to IF, CF significantly increased soil citrate-P and marginally decreased the HCl-P fractions, suggesting that the stable inorganic P pools are transferred to labile inorganic P at lower redox potentials. Compared to the -S treatments, + S treatments significantly increased the labile organic fractions (enzyme-P). Correspondingly, a decreased soil total organic P concentration was observed in + S treatment. Additionally, + S treatment significantly increased the activity of acid phosphomonoesterase and alkaline phosphomonoesterase and the abundance of phoD-harboring bacteria. These results indicated that straw promoted organic P minimization to release orthophosphate. The diversity of the phoD-harboring bacteria and complexity of the co-occurrence network decreased under the CF + S treatment; however, all keystone species of the phoD-harboring bacteria were retained in this oxygen-deficient environment. This study highlights that irrigation regimes mediate the processes of inorganic P mobilization, while straw returns regulate the processes of organic P mineralization. Additionally, flooding could be a more effective agricultural practice than straw returning to promote soil P availability in paddy soils. KEY POINTS: •Soil P pools and phoD-harboring bacteria communities were assessed. •Straw return mainly affects the mineralization of organic P. •Continuous flooding mainly affects the mobilization of inorganic P.

Phosphorus fractions and oxygen isotope composition of inorganic phosphate in typical agricultural soils.

Phosphorus (P), despite being an essential nutrient element for plants growth in agricultural ecosystem, the low utilization rate of soil P and the environmental problems caused by soil P losses are serious. Therefore, scoping knowledge of the possible sources and utilization extent of soil P by microorganisms is very helpful for better understanding of promoting P utilization for sustainable agriculture. Oxygen isotope of phosphate technology is an effective tool to trace the sources of P. In this study, P contents and oxygen isotope composition of inorganic phosphate (δ18OP) of different pools (H2O-P, NaHCO3-P, NaOH-P, and HCl-P) in typical agricultural soil from Northeast China and Central China were analyzed and quantified. The results showed that fertilizer and land use were important factors influencing the contents of H2O-Pt and NaHCO3-Pt and the soil TP contents from different types of soils were greatly affected by soil weathering degree. The δ18OP of different P pools indicated that the difference in utilization extent of different P fractions by microorganisms and the δ18OP values of different P fractions could be due to accumulation of multiple factors. The results will provide effective information for further study on sources and effective utilization of different P fractions in 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.

[Effects of Long-term Fertilization on Enzyme Activities in Profile of Paddy Soil Profiles].

The enzyme activity, which is closely related to soil material cycling (mineralization, transformation, etc.), can reflect soil quality and nutrient status. In order to explore the effect of long-term fertilization on the enzyme activity in paddy soil profile (0-40 cm), soils with organic fertilizer and inorganic fertilizer, and non-fertilized soils were selected, and the carbon and nitrogen contents, and the activities of ß-1,4-glucosidase (BG), and ß-1,4-N-acetylglucosaminidase (NAG) in 10cm depths of soil were analyzed. The results showed that the activities of BG and NAG in the soils treated with inorganic fertilizer and organic fertilizer increased by 0.73-47.87 nmol·(g·h)-1 and 1.33-128.81 nmol·(g·h)-1, and 0.19-9.72 nmol·(g·h)-1 and 0.92-57.66 nmol·(g·h)-1, respectively, compared to those for non-fertilized soil. Soil enzyme activity decreased with increasing soil depth. Soil enzyme activity in soil from 0-20 cm was significantly higher than that of soil from 20-40 cm. Soil enzyme activities were significantly affected by long term fertilization at different soil depths. RDA analysis showed that soil carbon and nitrogen contents had significant positive relationships with the activities of BG and NAG in the 0-20 cm soil profiles, however, negative relationships were observed in the 20-40 cm soil profiles. The long-term application of organic fertilizer significantly increased soil biomass and enzyme activity, both of which decreased with the increase in soil depth. Long-term fertilization could increase soil nutrient contents, microbial biomass, and extracellular enzyme activities, which has important theoretical significance for optimizing farmland fertilizer management and improving soil productivity.

[Effects of Long-term Fertilization Regimes on Microbial Biomass, Community Structure and Activity in a Paddy Soil].

Four paddy soils were collected in Ningxiang County, Hunan province. These used with different long-term fertilization regimes, including a control without fertilizer (CK), chemical fertilization with nitrogen, phosphate, and kalium (NPK), straw fertilization combined with NPK (ST), and manure fertilization combinedwith NPK (OM). Phospholipid fatty acid (PLFA) technology and MicrorespTM method were used to study the effect of long-term fertilization on soil microorganism abundance, community structure, and activity. Results showed that the abundance of bacteria, fungi, gram-negative (G-) bacteria, and gram-positive (G+) bacteria in the soil from the OM treatment was generally higher than for the other treatments; these levels were lower in the ST and NPK treatments and lowest in the CK treatment. The principal components analysis (PCA) of PLFA showed that the community structure of microorganisms in NPK, ST, and OM treatments was altered in comparison with that in CK, especially in the case of the ST and OM treatments. MicroRespTM results revealed that compared to the CK treatment (1.28 µg·h-1), soil microorganisms in the OM treatment had the highest average utilization rate of multiple carbon sources (1.81 µg·h-1), followed by ST (1.19 µg·h-1), CK (1.28 µg·h-1), and NPK (0.95 µg·h-1). Furthermore, different long-term fertilization regimes resulted in distinct carbon source preferences for the soil microorganisms and revealed a significant alteration in the microbial community. Conclusively, long-term fertilizer with straw or manure changes the microbial community and is a benefit for improving the biomass and activity of microorganism in rice paddy soils.

Effects of long-term fertilization on phoD-harboring bacterial community in Karst soils.

Phosphorus (P) acquisition by plants from soil organic P mainly relies on microorganisms. Examining the community of functional microbes that encode phosphatases (e.g. PhoD) under different fertilization managements may provide valuable information for promoting soil organic P availability. Here, we investigated how the abundance and community diversity of phoD-harboring bacteria responded to long-term fertilization in Karst soils. Six fertilization treatments were designed as follows: non-fertilized control (CK), inorganic fertilization only (NPK), and inorganic fertilization combined with low- and high amounts of straw (LSNPK and HSNPK), or cattle manure (LMNPK and HMNPK). We found that soil available phosphorus (AP) content and the activity of alkaline phosphatase (ALP) were significantly higher in all combined inorganic/organic fertilization treatments, while the abundance of the phoD gene was only higher in the HMPNK treatment, compared to NPK. The combination of inorganic/organic fertilizations had no effect on the diversity of phoD genes compared to NPK alone, but the phoD gene richness was greater in these treatments as compared to the control. Only organic fertilization combinations with high amounts of organic matter (both HSNPK and HMNPK) significantly affected the phoD community structure. A structure equation model demonstrated that soil organic carbon (SOC), rather than P, greatly affected the phoD community structure, suggesting that organic P mineralization in soils is decoupled from C mineralization. Our results suggested that optimized combinations of inorganic/organic fertilizations could promote P availability via regulating soil phoD-harboring bacteria community diversity and ALP activity.

[Effect of Phosphorus Addition on the Abundance of Autotrophic CO2-Fixation Microorganisms in Rhizospheric Soil from a Phosphorus-Limited Paddy Field].

A rice pot experiment was conducted to investigate the effect of phosphorus addition on the abundance of autotrophic CO2-fixation microorganisms using phosphorus-limited paddy soil from the Changsha Observation and Research Station for the Agricultural Environment. Rice seedlings were transplanted in the paddy soil with or without phosphorus addition, corresponding to P-treated-pot (P) or control pot (CK), respectively. Rhizosphere soils were collected from the P and CK treatments during the tillering and shooting stages. The physical and chemical soil properties were measured and the abundance of autotrophic CO2-fixation microorganisms was quantified with a real-time PCR technique based on four functional genes (cbbL, cbbM, accA, and aclB) involved in three CO2-fixation pathways (CBB cycle, rTCA cycle, and 3-hydroxypropionate/4-hydroxybutyrate cycle). The results show that phosphorus addition improves the concentrations of DOC and Olsen-P and the pH value, whereas negative effects on the MBC and NH4+-N concentrations are revealed during the tillering stage. The effect of phosphorus addition on the NO3--N concentration in the tillering and shooting stages differs. Phosphorus addition significantly increases the abundances of the cbbL, cbbM, accA, and aclB genes, which are 156%, 99%, 110%, and 193% higher than those of the CK treatment in the tillering stage. However, this positive effect is not notable for the cbbL, accA, and aclB genes during the shooting stage. Redundancy analysis (RDA) shows that Olsen-P is the environmental factor that most significantly affects the abundance of autotrophic CO2-fixation microorganisms.

[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.

Irrigation management and phosphorus addition alter the abundance of carbon dioxide-fixing autotrophs in phosphorus-limited paddy soil.

In this study, we assessed the interactive effects of phosphorus (P) application and irrigation methods on the abundances of marker genes (cbbL, cbbM, accA and aclB) of CO2-fixing autotrophs. We conducted rice-microcosm experiments using a P-limited paddy soil, with and without the addition of P fertiliser (P-treated-pot (P) versus control pot (CK)), and using two irrigation methods, namely alternate wetting and drying (AWD) and continuous flooding (CF). The abundances of bacterial 16S rRNA, archaeal 16S rRNA, cbbL, cbbM, accA and aclB genes in the rhizosphere soil (RS) and bulk soil (BS) were quantified. The application of P significantly altered the soil properties and stimulated the abundances of Bacteria, Archaea and CO2-fixation genes under CF treatment, but negatively influenced the abundances of Bacteria and marker genes of CO2-fixing autotrophs in BS soils under AWD treatment. The response of CO2-fixing autotrophs to P fertiliser depended on the irrigation management method. The redundancy analysis revealed that 54% of the variation in the functional marker gene abundances could be explained by the irrigation method, P fertiliser and the Olsen-P content; however, the rhizosphere effect did not have any significant influence. P fertiliser application under CF was more beneficial in improving the abundance of CO2-fixing autotrophs compared to the AWD treatment; thus, it is an ideal irrigation management method to increase soil carbon fixation.

[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.

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 selective microbial inhibitors on the microbial transformation of phosphorous in aggregates of highly weathered red soil with rice straw amendment].

In order to further understand the mechanisms of microbial immobilization of phosphorous (P) in highly weathered red soil with organic amendment, an incubation test was conducted to investigate the roles of microbial functional groups in the transformation of P in 0.2-2 mm soil aggregates. Throughout the 90-day incubation period, amendment with rice straw induced a substantial increase in the amounts of microbial biomass C and P, Olsen-P, and organic P in the aggregates. Comparing with rice straw amendment alone, the amendment with rice straw plus fungal inhibitor actidione decreased the amount of microbial biomass C in the aggregates by 10.5%-31.8% in the first 30 days. Such a decrement was significantly larger than that (6.8%-11.6%) in the treatment amended with rice straw plus bacterial inhibitors tetracycline and streptomycin sulphate (P<0.01). After the first 30 days, the microbial biomass C remained constant. In the first 20 days, the amount of microbial biomass P in the aggregates was 10.0%-28.8% higher in the treatment amended with bacterial inhibitors than in the treatment amended with fungal inhibitor (P<0.01). All the results suggested that that both the fungal and the bacterial groups were involved in the microbial immobilization of P in the soil aggregates, and the fungal group played a relatively larger role.