Microbial phosphorus-cycling genes in soil under global change.

The ongoing climate change on the Tibetan Plateau, leading to warming and precipitation anomalies, modifies phosphorus (P) cycling in alpine meadow soils. However, the interactions and cascading effects of warming and precipitation changes on the key "extracellular" and "intracellular" P cycling genes (PCGs) of bacteria are largely unknown for these P-limited ecosystems. We used metagenomics to analyze the individual and combined effects of warming and altered precipitation on soil PCGs and P transformation in a manipulation experiment. Warming and increased precipitation raised Olsen-P (bioavailable P, AP) by 13% and 20%, respectively, mainly caused by augmented hydrolysis of organic P compounds (NaOH-Po). The decreased precipitation reduced soil AP by 5.3%. The richness and abundance of the PCGs' community in soils on the cold Tibetan plateau were more sensitive to warming than altered precipitation. The abundance of PCGs and P cycling processes decreased under the influence of individual climate change factors (i.e., warming and altered precipitation alone), except for the warming combined with increased precipitation. Pyruvate metabolism, phosphotransferase system, oxidative phosphorylation, and purine metabolism (all "intracellular" PCG) were closely correlated with P pools under climate change conditions. Specifically, warming recruited bacteria with the phoD and phoX genes, which encode enzymes responsible for phosphoester hydrolysis (extracellular P cycling), strongly accelerated organic P mineralization and so, directly impacted P bioavailability in alpine soil. The interactions between warming and altered precipitation profoundly influenced the PCGs' community and facilitated microbial adaptation to these environmental changes. Warming combined with increased precipitation compensated for the detrimental impacts of the individual climate change factors on PCGs. In conclusion, warming combined with rising precipitation has boosting effect on most P-related functions, leading to the acceleration of P cycling within microbial cells and extracellularly, including mineralization and more available P release for microorganisms and plants in alpine soils.

Microbial strategies for phosphorus acquisition in rice paddies under contrasting water regimes: Multiple source tracing by 32P and 33P.

Microbial acquisition and utilization of organic and mineral phosphorus (P) sources in paddy soils are strongly dependent on redox environment and remain the key to understand P turnover and allocation for cell compound synthesis. Using double 32/33P labeling, we traced the P from three sources in a P-limited paddy soil: ferric iron-bound phosphate (Fe-P), wheat straw P (Straw-P), and soil P (Soil-P) in microbial biomass P (MBP) and phospholipids (Phospholipid-P) of individual microbial groups depending on water regimes: (i) continuous flooding or (ii) alternate wetting and drying. 32/33P labeling combined with phospholipid fatty acid analysis allowed to trace P utilization by functional microbial groups. Microbial P nutrition was mainly covered by Soil-P, whereas microorganisms preferred to take up P from mineralized Straw-P than from Fe-P dissolution. The main Straw-P mobilizing agents were Actinobacteria under alternating wetting and drying and other Gram-positive bacteria under continuous flooding. Actinobacteria and arbuscular mycorrhiza increased P incorporation into cell membranes by 1.4-5.8 times under alternate wetting and drying compared to continuous flooding. The Fe-P contribution to MBP was 4-5 times larger in bulk than in rooted soil because (i) rice roots outcompeted microorganisms for P uptake from Fe-P and (ii) rhizodeposits stimulated microbial activity, e.g. phosphomonoesterase production and Straw-P mineralization. Higher phosphomonoesterase activities during slow soil drying compensated for the decreased reductive dissolution of Fe-P. Concluding, microbial P acquisition strategies depend on (i) Soil-P, especially organic P, availability, (ii) the activity of phosphomonoesterases produced by microorganisms and roots, and (iii) P sources - all of which depend on the redox conditions. Maximizing legacy P utilization in the soil as a function of the water regime is one potential way to reduce competition between roots and microbes for P in rice cultivation.

Multi-imaging platform for rhizosphere studies: Phosphorus and oxygen fluxes.

Rhizosphere is a soil volume of high spatio-temporal heterogeneity and intensive plant-soil-microbial interactions, for which visualization and process quantification is of highest scientific and applied relevance, but still very challenging. A novel methodology for quick assessment of two-dimensional distribution of available phosphorus (P) in rhizosphere was suggested, tested, and development up to the application platform. Available P was firstly trapped by an in-situ diffusive gradients in thin-films (DGT) sampler with precipitated zirconia as the binding gel, and subsequently, the loaded gel was analyzed with an optimized colorimetric imaging densitometry (CID). The imaging platform was established linking: i) DGT, ii) planar optode, and iii) soil zymography techniques to simultaneously determine available P, oxygen, and acid phosphatase in rhizosphere at sub-millimeter spatial scales. The DGT identified available P level in rice rhizosphere were spatially overlapping to the localized redox hotspots and phosphatase activity. The spatial relationship between available P and acid phosphatase activity was dependent on root development. The root radial oxygen loss (ROL) remained active during the experimental observations (2-3 days), while a flux of available P of 10 pg cm-2 s-1 was visualized within 2-3 mm of roots, confirming the correlative response of rice roots to oxygen secretion and P uptake. Summarizing, the established imaging platform is suitable to capture spatial heterogeneity and temporal dynamics of root activities, nutrient bioavailability, ROL and enzyme activities in rhizosphere.

Long-term nitrogen and phosphorus fertilization reveals that phosphorus limitation shapes the microbial community composition and functions in tropical montane forest soil.

Microorganisms govern soil nutrient cycling. It is therefore critical to understand their responses to human-induced increases in N and P inputs. We investigated microbial community composition, biomass, functional gene abundance, and enzyme activities in response to 10-year N and P addition in a primary tropical montane forest, and we explored the drivers behind these effects. Fungi were more sensitive to nutrient addition than bacteria, and the fungal community shift was mainly driven by P availability. N addition aggravated P limitation, to which microbes responded by increasing the abundance of P cycling functional genes and phosphatase activity. In contrast, P addition alleviated P deficiency, and thus P cycling functional gene abundance and phosphatase activity decreased. The shift of microbial community composition, changes in functional genes involved in P cycling, and phosphatase activity were mainly driven by P addition, which also induced the alteration of soil stoichiometry (C/P and N/P). Eliminating P deficiency through fertilization accelerated C cycling by increasing the activity of C degradation enzymes. The abundances of C and P functional genes were positively correlated, indicating the intensive coupling of C and P cycling in P-limited forest soil. In summary, a long-term fertilization experiment demonstrated that soil microorganisms could adapt to induced environmental changes in soil nutrient stoichiometry, not only through shifts of microbial community composition and functional gene abundances, but also through the regulation of enzyme production. The response of the microbial community to N and P imbalance and effects of the microbial community on soil nutrient cycling should be incorporated into the ecosystem biogeochemical model.

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.

Phosphorus addition decreases plant lignin but increases microbial necromass contribution to soil organic carbon in a subalpine forest.

Increasing phosphorus (P) inputs induced by anthropogenic activities have increased P availability in soils considerably, with dramatic effects on carbon (C) cycling and storage. However, the underlying mechanisms via which P drives plant and microbial regulation of soil organic C (SOC) formation and stabilization remain unclear, hampering the accurate projection of soil C sequestration under future global change scenarios. Taking the advantage of an 8-year field experiment with increasing P addition levels in a subalpine forest on the eastern Tibetan Plateau, we explored plant C inputs, soil microbial communities, plant and microbial biomarkers, as well as SOC physical and chemical fractions. We found that continuous P addition reduced fine root biomass, but did not affect total SOC content. P addition decreased plant lignin contribution to SOC, primarily from declined vanillyl-type phenols, which was coincided with a reduction in methoxyl/N-alkyl C by 2.1%-5.5%. Despite a decline in lignin decomposition due to suppressed oxidase activity by P addition, the content of lignin-derived compounds decreased because of low C input from fine roots. In contrast, P addition increased microbial (mainly fungal) necromass and its contribution to SOC due to the slower necromass decomposition under reduced N-acquisition enzyme activity. The larger microbial necromass contribution to SOC corresponded with a 9.1%-12.4% increase in carbonyl C abundance. Moreover, P addition had no influence on the slow-cycing mineral-associated organic C pool, and SOC chemical stability indicated by aliphaticity and recalcitrance indices. Overall, P addition in the subalpine forest over 8 years influenced SOC composition through divergent alterations of plant- and microbial-derived C contributions, but did not shape SOC physical and chemical stability. Such findings may aid in accurately forecasting SOC dynamics and their potential feedbacks to climate change with future scenarios of increasing soil P availability in Earth system models.

Shrubs magnify soil phosphorus depletion in Tibetan meadows: Conclusions from C:N:P stoichiometry and deep soil profiles.

Globally, the proliferation of shrubs within grasslands stimulates soil phosphorus (P) cycling and increases topsoil P storage beneath their canopies. However, little is known regarding the impact of shrub encroachment on subsoil P storage, and whether shrubs mediate changes in soil stoichiometry, like increasing P cycling. In grazed meadows on the Tibetan Plateau, soil and roots were sampled to 1 m depth in shrubby Hippophae rhamnoides ssp. sinensis groves and the surrounding grassy areas. Shrubs had higher P content than grasses, but lower C:P ratios in their leaves, litter, and roots. Similarly, shrubs had higher microbial P content than grasses, but lower microbial biomass C:P and N:P ratios in the soil. The larger microbial P stock in the 1 m of soil beneath shrubs responded to the larger root P stock there as well. Thus, both the plants and microbes acquired more P in shrubby areas than in grassy areas by accelerating P mineralization. The greater net production of available P in the topsoil and the synthesis of microbial P throughout the profile under shrubs increased the P solubility. Total P, inorganic P, and organic P stocks were lower under shrubs than under grasses in the top 1 m of soil. This decrease in soil P storage beneath shrubs is most likely attributable to P leaching due to higher P solubility, heavy rainfall, and larger soil gaps. Moreover, shrubs also had larger plant biomass P stock compared to grasses, and thus the depletion of P from the top 1 m of soil was further magnified via plant biomass removal. We concluded that shrubs increase P cycling to overcome the stoichiometric imbalance between their P requirement and the supply in the soil, and the fast P cycling under shrubs magnify P depletion within the rooted soil depth in alpine meadows.

Allocation of freshly assimilated carbon into primary and secondary metabolites after in situ ¹³C pulse labelling of Norway spruce (Picea abies).

Plants allocate carbon (C) to sink tissues depending on phenological, physiological or environmental factors. We still have little knowledge on C partitioning into various cellular compounds and metabolic pathways at various ecophysiological stages. We used compound-specific stable isotope analysis to investigate C partitioning of freshly assimilated C into tree compartments (needles, branches and stem) as well as into needle water-soluble organic C (WSOC), non-hydrolysable structural organic C (stOC) and individual chemical compound classes (amino acids, hemicellulose sugars, fatty acids and alkanes) of Norway spruce (Picea abies) following in situ (13)C pulse labelling 15 days after bud break. The (13)C allocation within the above-ground tree biomass demonstrated needles as a major C sink, accounting for 86% of the freshly assimilated C 6 h after labelling. In needles, the highest allocation occurred not only into the WSOC pool (44.1% of recovered needle (13)C) but also into stOC (33.9%). Needle growth, however, also caused high (13)C allocation into pathways not involved in the formation of structural compounds: (i) pathways in secondary metabolism, (ii) C-1 metabolism and (iii) amino acid synthesis from photorespiration. These pathways could be identified by a high (13)C enrichment of their key amino acids. In addition, (13)C was strongly allocated into the n-alkyl lipid fraction (0.3% of recovered (13)C), whereby (13)C allocation into cellular and cuticular exceeded that of epicuticular fatty acids. (13)C allocation decreased along the lipid transformation and translocation pathways: the allocation was highest for precursor fatty acids, lower for elongated fatty acids and lowest for the decarbonylated n-alkanes. The combination of (13)C pulse labelling with compound-specific (13)C analysis of key metabolites enabled tracing relevant C allocation pathways under field conditions. Besides the primary metabolism synthesizing structural cell compounds, a complex network of pathways consumed the assimilated (13)C and kept most of the assimilated C in the growing needles.

Microbial growth and carbon use efficiency in the rhizosphere and root-free soil.

Plant-microbial interactions alter C and N balance in the rhizosphere and affect the microbial carbon use efficiency (CUE)-the fundamental characteristic of microbial metabolism. Estimation of CUE in microbial hotspots with high dynamics of activity and changes of microbial physiological state from dormancy to activity is a challenge in soil microbiology. We analyzed respiratory activity, microbial DNA content and CUE by manipulation the C and nutrients availability in the soil under Beta vulgaris. All measurements were done in root-free and rhizosphere soil under steady-state conditions and during microbial growth induced by addition of glucose. Microorganisms in the rhizosphere and root-free soil differed in their CUE dynamics due to varying time delays between respiration burst and DNA increase. Constant CUE in an exponentially-growing microbial community in rhizosphere demonstrated the balanced growth. In contrast, the CUE in the root-free soil increased more than three times at the end of exponential growth and was 1.5 times higher than in the rhizosphere. Plants alter the dynamics of microbial CUE by balancing the catabolic and anabolic processes, which were decoupled in the root-free soil. The effects of N and C availability on CUE in rhizosphere and root-free soil are discussed.