Plant and microbial uptake of nitrogen and phosphorus affected by drought using 15N and 32P tracers

Competition for nutrients between plants and microbes is an important determinant for plant growth, biodiversity and carbon cycling. Perturbations such as drought affect the availability of nitrogen (N) and phosphorus (P), and may cause shifts in uptake of N and P between plants and microbes. Competitiveness for these nutrients may depend on how flexible plants and microbes are in taking up N and P. We used a novel dual isotope labelling technique (¹⁵N and ³²P) to assess short-term uptake of N and P by plants and microbes affected by drought in two different plant–soil systems. Mesocosms were extracted from two grassland sites differing in soil nutrient availability and plant species. Half of the mesocosms were subjected to drought one week prior to injection of ¹⁵N (as KNO₃) and ³²P (as H₃PO₄) tracers. Uptake rates of ${{\text{NO}}_3}^{-}$ and P in plants and microbes were estimated based on average source pool enrichment during the labelling period and on plant and microbial recovery of ¹⁵N and ³²P measured after 4 days of labelling. Overall competition for N and P was reduced with drought as less ${{\text{NO}}_3}^{-}$ and P was taken up in plants and microbes. However, plant uptake of ${{\text{NO}}_3}^{-}$ was more sensitive to drought than microbial ${{\text{NO}}_3}^{-}$ uptake, while microbial P uptake was more sensitive than plant P uptake. These different sensitivities to drought by plants and microbes may decouple the N and P cycle with increased drought conditions.     

毒死蜱对棉花根际土细菌群落多样性和结构的影响

通过室内盆栽试验模拟自然环境条件,采用高效液相色谱(HPLC)和末端限制性片段长度多态性(T-RFLP)技术,研究了土壤使用推荐剂量(5 mg·kg~(-1))及推荐剂量的2倍、3倍和4倍(10 mg·kg~(-1)、15 mg·kg~(-1)、20 mg·kg~(-1))毒死蜱对棉花根际土壤细菌群落多样性和结构的影响,以不施用毒死蜱的土壤为对照。结果表明,5 mg·kg~(-1)、10 mg·kg~(-1)、15 mg·kg~(-1)和20 mg·kg~(-1)毒死蜱在土壤中的半衰期分别为10.04 d、11.36 d、11.55 d和12.16 d,60 d时基本完全降解。毒死蜱处理60 d后,棉花生物量显著降低;毒死蜱浓度越高,棉花生物量越低。无毒死蜱条件下不同取样时间根际细菌多样性无显著差异,毒死蜱处理组前30 d细菌多样性均显著降低,60 d时毒死蜱处理组细菌多样性恢复到正常水平。研究发现毒死蜱浓度越高对细菌多样性抑制作用越显著,恢复越缓慢。主成分分析结果发现,第10 d、30 d和60 d毒死蜱处理组与对照组细菌群落结构差异显著,其中60 d时20 mg·kg~(-1)毒死蜱处理组差异最显著,即使土壤中毒死蜱完全降解,根际细菌群落结构仍不会恢复到正常水平。60 d时,被毒死蜱抑制的细菌有硝化刺菌属(Nitrospina sp.)和Cellulophaga sp.等,被激活的有芽孢杆菌属(Bacillus sp.)和链霉菌属(Streptomyces sp.)等。可见,毒死蜱的引入,重新构建了土壤细菌群落结构,显著影响棉花生长,对棉花根际土壤微生态环境冲击较大,应对其生态安全性予以重视。     

Measuring microbial uptake of nitrogen in nutrient-amended sandy soils—A mass-balance based approach

Soil microbial biomass N is commonly determined through fumigation-extraction (FE), and a conversion factor (K_EN) is necessary to convert extractable N to actual soil biomass N. Estimation of K_EN has been constrained by various uncertainties including potential microbial immobilisation. We developed a mass-balance approach to quantify changes in microbial N storage during nutrient-amended incubation, in which microbial uptake is determined as the residual in a ‘mass-balance’ based on soil-water N before and after amended incubation. The approach was applied to three sandy soils of southwestern Australia, to determine microbial N immobilisation during 5-day incubation in response to supply of 2.323 mg C g⁻¹, 100 μg N g⁻¹ and 20 μg P g⁻¹. The net N immobilisation was estimated to be 95-114 μg N g⁻¹ in the three soils, equivalent to 82.7-85.1% of soil-water N following the amendment. Such estimation for microbial uptake does not depend on fumigation and K_EN conversion, but for comparison purposes we estimated ‘nominal’ K_EN values (0.11-0.14) for the three soils, which were comparable to previously reported K_EN from soils receiving C and N amendment. The accuracy of our approach depends on the mass-balance equation and the integrated measurement errors of the multiple N pools, and was assessed practically through recoveries of added-N when microbial uptake can be minimised. Near-satisfactory recoveries were achieved under such conditions. Our mass-balance approach provides information not only about changes in the microbial biomass nitrogen storage, but also major N-pools and their fluxes in regulating soil N concentrations under substrate and nutrient amended conditions.     

Comparison of the difference method and 15N technique for studying the fate of nitrogen from plant residues in soil

We compared the dynamics of net mineralization of nitrogen (N) derived from white clover material (Ndfc) as measured by the difference and the ¹⁵N methods in a pot experiment with a sandy loam (15°C and pF 2.4) planted with Italian ryegrass. On day 22, mineralized Ndfc (soil mineral N plus plant N uptake) was 5.8% and 1.3% of added N for the ¹⁵N and the difference methods, respectively. The discrepancy was reduced on day 43. On day 64, the relationship was reversed, and on day 98 the values given by the two methods were 22.8% and 29.5%, respectively. The results obtained by the two methods were linearly correlated (r = 0.987) and, on average, did not differ significantly. Nevertheless, the different temporal patterns led to appreciably different parameter values as estimated by fitting of a reparameterized Richards model. On day 22, clover amendment reduced mineralized N derived from soil (Ndfs) by 3.4 mg N pot^–1. The reason for this was that the clover amendment led to a reduction in plant growth and uptake of Ndfs, most likely because of allelopathy, while mineral Ndfs did not increase correspondingly. Clover-induced Ndfs in the microbial biomass of 5.1 mg N pot^–1 suggested that the mineral Ndfs not taken up by plants had been reimmobilized. Towards the end of the experiment, clover-induced Ndfs in the biomass declined to 1.5 mg N pot^–1, while mineralized Ndfs due to clover amendment increased to 5.1 mg N pot^–1. The results strongly suggested that this increase was caused by a real stimulation of humus N mineralization by clover amendment rather than by isotope displacement or pool substitution. Received: 5 May 1997     

Plant biomass influences rhizosphere priming effects on soil organic matter decomposition in two differently managed soils

We used a continuous labeling method of naturally ¹³C-depleted CO₂ in a growth chamber to test for rhizosphere effects on soil organic matter (SOM) decomposition. Two C3 plant species, soybean (Glycine max) and sunflower (Helianthus annus), were grown in two previously differently managed soils, an organically farmed soil and a soil from an annual grassland. We maintained a constant atmospheric CO₂ concentration at 400±5 ppm and δ¹³C signature at −24.4‰ by regulating the flow of naturally ¹³C-depleted CO₂ and CO₂-free air into the growth chamber, which allowed us to separate new plant-derived CO₂-C from original soil-derived CO₂-C in soil respiration. Rhizosphere priming effects on SOM decomposition, i.e., differences in soil-derived CO₂-C between planted and non-planted treatments, were significantly different between the two soils, but not between the two plant species. Soil-derived CO₂-C efflux in the organically farmed soil increased up to 61% compared to the no-plant control, while the annual grassland soil showed a negligible increase (up to 5% increase), despite an overall larger efflux of soil-derived CO₂-C and total soil C content. Differences in rhizosphere priming effects on SOM decomposition between the two soils could be largely explained by differences in plant biomass, and in particular leaf biomass, explaining 49% and 74% of the variation in primed soil C among soils and plant species, respectively. Nitrogen uptake rates by soybean and sunflower was relatively high compared to soil C respiration and associated N mineralization, while inorganic N pools were significantly depleted in the organic farm soil by the end of the experiment. Despite relatively large increases in SOM decomposition caused by rhizosphere effects in the organic farm soil, the fast-growing soybean and sunflower plants gained little extra N from the increase in SOM decomposition caused by rhizosphere effects. We conclude that rhizosphere priming effects of annual plants on SOM decomposition are largely driven by plant biomass, especially in soils of high fertility that can sustain high plant productivity.     

The influence of two agricultural biostimulants on nitrogen transformations, microbial activity, and plant growth in soil microcosms

The influence of two experimental soil treatments, Z93 and W91, on nitrogen transformations, microbial activity and plant growth was investigated in soil microcosms. These compounds are commercially marketed fermentation products (Agspectrum) that are sold to be added to field soils in small amounts to promote nitrogen and other nutrient uptake by crops in USA. In laboratory microcosm experiments, soils were amended with finely ground alfalfa-leaves or wheat straw, or left unamended, in an attempt to alter patterns of soil nitrogen mineralization and immobilization. Soils were treated in the microcosms with Z93 and W91 at rates equivalent to the recommended field application rates, that range from 0.2 to 1.1 l ha⁻¹, (0.005-0.03 μl g⁻¹ soil). We measured their effects on soil microbial activity (substrate-induced respiration (SIR), dehydrogenase activity (DHA) and acid phosphatase activity (PHOS)), soil nitrogen pools (microbial biomass N, mineral N, dissolved organic N), and transformations (net N mineralization and nitrification, ¹⁵N dilution of the mineral N pool, and accumulation of mineral N on ion-exchange resins), and on wheat plant germination and growth (shoot and root biomass, shoot length, N uptake and ¹⁵N enrichment of shoot tissues), for up to 56 days after treatment. To follow the movement of nitrogen from inorganic fertilizer into plant biomass we used a ¹⁵N isotopic tracer. Most of the soil and plant responses to treatment with Z93 or W91 differed according to the type of organic amendment that was used. Soil treatment with either Z93 or W91 influenced phosphatase activity strongly but did not have much effect on SIR or DHA. Both chemicals altered the rates of decomposition and mineralization of organic materials in the soil, which was evidenced by significant increases in the rates of the decomposition of buried wheat straw, and by the acceleration of net, rates of N mineralization, relative to those of the controls. Soil nitrate availability increased at the end of the experiment in response to both chemical treatments. In alfalfa-amended soils, the final plant biomass was decreased significantly by treatment with W91. Increased plant growth and N-use efficiency in straw-amended soil, resulting from treatments with Z93 or W91, was linked to increased rates of N mineralization from indigenous soil organic materials. This supports the marketing of these compounds as promoters of N uptake at these low dosage inputs.     

氯化钠不同浓度对夏玉米生长和吸氮的影响

采用珍珠岩为基质的温室盆栽试验,研究不同盐分(NaCl)水平对夏玉米生长和吸氮的影响。试验设灌溉水中NaCl浓度为0、10、20、40和60mmol.L5个处理。结果表明,在出苗后的25～101d(7月25日～10月10日),当NaCl浓度达到60mmol.L时,明显降低了夏玉米的叶面积、株高、干物质量和根长;盐分对植株体内各器官的氮含量影响不大。随着NaCl含量的增加,植株的总氮累积量将减少,且植株中累积氮量随时间呈二次函数增长。     

Soil microbial metabolism and nutrient status in a Mediterranean area as affected by plant cover

The Mediterranean area of Southern Italy is characterized by different natural plant covers that mainly reflect different successional stages (i.e. low maquis, high maquis, Quercus ilex wood) and managed areas with introduced plant species (such as Pinus species). Soil properties could be affected by plant cover types as well as by plant species. Our objective was to determine the relationships of plant cover types and plant species with the chemical and biological characteristics of the soil. In four neighbouring areas with different plant cover types (low maquis, pure high maquis, high maquis with pines and pinewood, with pines planted by foresters in both cases), soil samples were collected under different plant species in order to evaluate the effect of plant cover types and plant species on soil properties. Soil samples were analyzed for nutrient content, microbial biomass, soil potential respiration and enzymatic activity (phosphatase, arylsulphatase, β-glucosidase and hydrolase activities) as well as for pH, water holding capacity (WHC) and cation exchange capacity (CEC). Application of cluster analysis and principal component analysis to the data revealed that the plant cover type was the key factor influencing soil properties more than plant species. In fact, the largest differences were observed between pure high maquis soils and all other soils, with pure high maquis soils generally showing the highest values of WHC, CEC, nutrient content, organic and microbial C, soil respiration, phosphatase, arylsulphatase and β-glucosidase activities. The significantly lower values of these variables in the low maquis relative to the pure high maquis probably reflect the effect of ecological succession on soil. The high maquis with pine, differing from the pure high maquis only for the presence or absence of pine, showed values of soil physical, chemical and biological characteristics similar to those found in the low maquis, thus suggesting that the presence of pine retards soil development.     

氮钾水互作对玉米苗期植株生长及钾素吸收的影响

采用盆栽试验方法探讨了氮钾水互作对玉米苗期植株生长及养分吸收的影响。结果表明,水分适宜能明显增加玉米植株株高和干物重,水分适宜条件下玉米株高和干物重较水分亏缺条件的分别增加7.8%和13.8%。增施氮肥能显著增加玉米植株株高和干物重,水分适宜条件下中氮水平的玉米株高和干物重分别较低氮水平的增加10.4%和8.7%,而水分亏缺条件下株高和干物重均随施氮水平的增加而明显增加;水分亏缺条件下,中高量施钾能显著增加玉米植株干物重。水分适宜条件下增施氮肥能明显促进玉米对钾素的吸收,在水分适宜和亏缺条件下,不同氮水平的玉米钾素吸收均随施钾水平的增加而显著增加。     

Effect of microbial nitrogen immobilization during the growth period on the availability of nitrogen fertilizer for winter cereals

 Pot and field experiments were conducted to determine microbial immobilization of N fertilizer during growth periods of winter wheat and winter barley. In a pot experiment with winter wheat, Ca(¹⁵NO₃)₂ was applied at tillering [Zadok's growth stage (GS) 25)], stem elongation (GS 31) and ear emergence (GS 49). Rates of 100 mg N pot^–1, 200 mg N pot^–1 or 300 mg N pot^–1 were applied at each N application date. At crop maturity, ¹⁵N-labelled fertilizer N immobilization was highest at the highest N rate (3×300 mg N pot^–1). For each N-rate treatment about 50% of the total immobilized fertilizer N was immobilized from the first N dressing, and 30% and 20% of the total ¹⁵N immobilized was derived from the second and third applications, respectively. In field trials with winter wheat (three sites) and winter barley (one site) N was applied at the same growth stages as for the pot trial. N was also applied to fallow plots, but only at GS 25. N which was not recovered (neither in crops nor in soil mineral N pools) was considered to represent net immobilized N. A clear effect of N rate (51–255 kg N ha^–1) on net N immobilization was not found. The highest net N immobilization was found for the period between GS 25 (March) and GS 31 (late April) which amounted to 54–97% of the total net N immobilized at harvest (July/August). At GS 31, non-recovered N was found to be of similar magnitude for cropped and fallow plots, indicating that C from roots did not affect net N immobilization. Microbial biomass N (N_mic) was determined for cropped plots at GS 31. Although N_mic tended to be higher in fertilized than in unfertilized plots, fertilizer-induced increases in N_mic and net N immobilization were poorly correlated. It can be concluded that microbial immobilization of fertilizer N is particularly high after the first N application when crop growth and N uptake are low. Received: 6 July 1999     

Additive and interactive effects of functionally dissimilar soil organisms on a grassland plant community

The productivity and diversity of plant communities are affected by soil organisms such as arbuscular mycorrhizal fungi (AMF), root herbivores and decomposers. However, it is unknown how interactions between such functionally dissimilar soil organisms affect plant communities and whether the combined effects are additive or interactive. In a greenhouse experiment we investigated the individual and combined effects of AMF (five Glomus species), root herbivores (wireworms and nematodes) and decomposers (collembolans and enchytraeids) on the productivity and nutrient content of a model grassland plant community as well as on soil microbial biomass and community structure. The effects of the soil organisms on productivity (total plant biomass), total root biomass, grass and forb biomass, and nutrient uptake of the plant community were additive. AMF decreased, decomposers increased and root herbivores had no effect on productivity, but in combination the additive effects canceled each other out. AMF reduced total root biomass by 18%, but decomposers increased it by 25%, leading to no net effect on total root biomass in the combined treatments. Total shoot biomass was reduced by 14% by root herbivores and affected by an interaction between AMF and decomposers where decomposers had a positive impact on shoot growth only in presence of AMF. AMF increased the shoot biomass of forbs, but reduced the shoot biomass of grasses, while root herbivores only reduced the shoot biomass of grasses. Interactive effects of the soil organisms were detected on the shoot biomasses of Lotus corniculatus, Plantago lanceolata, and Agrostis capillaris. The C/N ratio of the plant community was affected by AMF.In soil, AMF promoted abundances of bacterial, actinomycete, saprophytic and AMF fatty acid markers. Decomposers alone decreased bacterial and actinomycete fatty acids abundances but when decomposers were interacting with herbivores those abundances were increased. Our results suggests that at higher resolutions, i.e. on the levels of individual plant species and the microbial community, interactive effects are common but do not affect the overall productivity and nutrient uptake of a grassland plant community, which is mainly affected by additive effects of functionally dissimilar soil organisms.     

双季稻田冬闲期土壤细菌数量及结构对施氮的响应

[目的]研究不同施氮水平对湖南双季稻区冬闲期土壤细菌结构与数量的影响,为双季稻区水稻可持续生产提供理论依据。[方法]依托湖南省双季稻区连续8年的定位试验,选取3个氮肥水平处理:CK(不施氮肥)、N1(施N 100 kg/hm^2)、N2(施N 200 kg/hm^2),取稻田冬闲期5—20 cm耕层土样,采用高通量测序和荧光定量PCR方法测定了土壤细菌数量与结构。[结果]与CK相比,N1、N2处理显著增加了双季稻产量,提升了冬闲期土壤总氮、全碳含量,降低了土壤pH、硝态氮及碳氮比(P<0.05)。N1和N2处理的细菌16s rDNA基因拷贝数分别为CK的48.25和40.31倍。施氮显著增加土壤总细菌丰度,土壤细菌丰度与土壤全氮、碳氮比呈显著正相关,与土壤全碳呈显著负相关。施氮改变了冬闲期稻田土壤细菌的多样性及群落结构。与CK相比,N1处理提高了稻田土壤微生物多样性;N2处理显著增加稻田土壤细菌丰富度,但显著降低稻田土壤细菌多样性。此外,3个处理土壤菌群相对丰度最高的是Proteobacteria(变形菌门),达40.16%~51.16%。N2处理的变形菌门相对丰度显著高于CK与N1,N1处理的变形菌门相对丰度显著低于CK。3个处理属水平细菌相对丰度较高的是Anaerolineaceae_uncultured(厌氧绳菌属)和Nitrospira(硝化螺旋菌属),其相对丰度分别为8.6%~14.56%和8.16%~11.46%。Spearman相关性分析显示,11个优势门菌群数量均与土壤化学性质存在显著相关性。稻田11个优势菌群的数量与土壤化学性质显著相关。[结论]湖南双季稻区施氮降低冬闲期稻田土壤pH和C/N比,低施氮水平可增加稻田微生物多样性,高施氮量虽然增加稻田细菌丰富度,但降低了微生物多样性。     

Production of root-derived material and associated microbial growth in soil at different nutrient levels

Summary Maize plants were grown for 42 days in a sandy soil at two different mineral nutrient levels, in an atmosphere containing ¹⁴CO₂. The ¹⁴C and total carbon contents of shoots, roots, soil and soil microbial biomass were measured 28, 35 and 42 days after germination. Relative growth rates of shoots and roots decreased after 35 days at the lower nutrient level, but were relatively constant at the higher nutrient level. In the former treatment, 2% of the total ¹⁴C fixed was retained as a residue in soil at all harvests while at the higher nutrient level up to 4% was retained after 42 days. Incorporation of ¹⁴C into the soil microbial biomass was close to its maximum after 35 days at the lower nutrient level, but continued to increase at the higher level. Generally a good agreement existed between microbial biomass, ¹⁴C contents and numbers of fluorescent pseudomonads in the rhizosphere. Numbers of fluorescent pseudomonads in the rhizosphere were maximal after 35 days at the lower nutrient level and continued to increase at the higher nutrient level. The proportions of the residual ¹⁴C in soil, incorporated in the soil microbial biomass, were 28% to 41% at the lower nutrient level and 20%6 – 30% at the higher nutrient level. From the lower nutrient soil 18%6 – 52%6 of the residual soil ¹⁴C could be extracted with 0.5 N K₂SO₄, versus 14%6 – 16% from the higher nutrient soil.Microbial growth in the rhizosphere seemed directly affected by the depletion of mineral nutrients while plant growth and the related production of root-derived materials continued.     

Azoarcus sp. strain KH32C affects rice plant growth and the root-associated soil bacterial community in low nitrogen input paddy fields

ABSTRACT

Excessive input of nitrogen fertilizer causes nitrogen pollution in aquatic environments. Utilizing microbial inoculants seems to be effective in decreasing the extent of nitrogen fertilizer application. Genome sequencing analysis of Azoarcus sp. strain KH32C, which was isolated from a paddy-upland rotation paddy field in Japan, showed that the strain may interact with plants. In this study, we examined the effect of inoculation of rice seeds with strain KH32C as a bacterial inoculant in paddy fields without nitrogen fertilization. Rice of three cultivars inoculated with strain KH32C were cultivated in paddy fields with different soil carbon and nitrogen levels. We investigated the growth rate of rice plants and the elemental composition of brown rice. In addition, the bacterial community structures in the rice root-associated soil were examined using molecular genetic analysis. KH32C inoculation resulted in an increase in the rice plant growth rate in the early growth phase with cultivar Nipponbare. Elemental composition analysis showed that the zinc concentration in brown rice of Nipponbare was increased by KH32C inoculation. KH32C inoculation affected the bacterial community in Nipponbare root-associated soil. The community of potential plant growth-promoting bacteria, the majority of which were in the class Alphaproteobacteria, had relatively high abundance in the early growth stage after KH32C inoculation. Our results indicated that strain KH32C can be utilized as a bacterial inoculant with the effect of promoting rice growth and enhancing the zinc content of rice grains in low nitrogen input paddy fields.     

Water limitation and plant inter-specific competition reduce rhizosphere-induced C decomposition and plant N uptake

Plants can affect soil organic matter decomposition and mineralization through litter inputs, but also more directly through root-microbial interactions (rhizosphere effects). Depending on resource availability and plant species identity, these rhizosphere effects can be positive or negative. To date, studies of rhizosphere effects have been limited to plant species grown individually. It is unclear how belowground resources and inter-specific interactions among plants may influence rhizosphere effects on soil C decomposition and plant N uptake. In this study, we tested the simple and interactive effects of plant diversity and water availability on rhizosphere-mediated soil C decomposition and plant N uptake. The study was conducted in the greenhouse with five semi-arid grassland species (monocultures and mixtures of all five species) and two water levels (15 and 20% gravimetric soil moisture content). We hypothesized that microbial decomposition and N release would be less in the low compared to high water treatment and less in mixtures compared to monocultures. Rhizosphere effects on soil C decomposition were both positive and negative among the five species when grown in monoculture, although negative effects prevailed by the end of the experiment. When grown in mixture, rhizosphere effects reduced soil C decomposition and plant N uptake compared to monocultures, but only at the low-water level. Our results suggest that when water availability is low, plant species complementarity and selection effects on water and N use can decrease soil C decomposition through rhizosphere effects. Although complementarity and selection effects can increase plant N uptake efficiency, plant N uptake in the mixtures was still lower than expected, most likely because rhizosphere effects reduced N supply in the mixtures more than in the monocultures. Our results indicate that rhizosphere effects on C and N cycling depend on water availability and inter-specific plant interactions. Negative rhizosphere effects on soil C decomposition and N supply in mixtures relative to monocultures of the component species could ultimately increase soil C storage and possibly influence how plant communities in semi-arid grasslands respond to global climate change.