Modeling soil metabolic processes using isotopologue pairs of position-specific C-labeled glucose and pyruvate

Most organic carbon (C) in soils eventually turns into CO₂ after passing through microbial metabolic pathways, while providing cells with energy and biosynthetic precursors. Therefore, detailed insight into these metabolic processes may help elucidate mechanisms of soil C cycling processes. Here, we describe a modeling approach to quantify the C flux through metabolic pathways by adding 1-¹³C and 2,3-¹³C pyruvate and 1-¹³C and U-¹³C glucose as metabolic tracers to intact soil microbial communities. The model calculates, assuming steady-state conditions and glucose as the only substrate, the reaction rates through glycolysis, Krebs cycle, pentose phosphate pathway, anaplerotic activity through pyruvate carboxylase, and various biosynthesis reactions. The model assumes a known and constant microbial proportional precursor demand, estimated from literature data. The model is parameterized with experimentally determined ratios of ¹³CO₂ production from pyruvate and glucose isotopologue pairs. Model sensitivity analysis shows that metabolic flux patterns are especially responsive to changes in experimentally determined ¹³CO₂ ratios from pyruvate and glucose. Calculated fluxes are far less sensitive to assumptions concerning microbial chemical and community composition. The calculated metabolic flux pattern for a young volcanic soil indicates significant pentose phosphate pathway activity in excess of pentose precursor demand and significant anaplerotic activity. These C flux patterns can be used to calculate C use efficiency, energy production and consumption for growth and maintenance purposes, substrate consumption, nitrogen demand, oxygen consumption, and microbial C isotope composition. The metabolic labeling and modeling methods may improve our ability to study the biochemistry and ecophysiology of intact and undisturbed soil microbial communities.     

Short term soil priming effects and the mineralisation of biochar following its incorporation to soils of different pH

The aim of this work was to determine the magnitude of the priming effect, i.e. short-term changes in the rate (negative or positive) of mineralisation of native soil organic carbon (C), following addition of biochars. The biochars were made from Miscanthus giganteus, a C4 plant, naturally enriched with ¹³C. The biochars were produced at 350 °C (biochar350) and 700 °C (biochar700) and applied with and without ryegrass as a substrate to a clay-loam soil at pH 3.7 and 7.6. A secondary aim was to determine the effect of ryegrass addition on the mineralisation of the two biochars.After 87 days, biochar350 addition caused priming effects equivalent to 250 and 319 μg CO₂-C g⁻¹ soil, in the low and high pH soil, respectively. The largest priming effects occurred at the start of the incubations. The size of the priming effect was decreased at higher biochar pyrolysis temperatures, which may be a way of controlling priming effects following biochar incorporation to soil, if desired. The priming effect was probably induced by the water soluble components of the biochar. At 87 days of incubation, 0.14% and 0.18% of biochar700 and 0.61% and 0.84% of biochar350 were mineralized in the low and high pH soil, respectively. Ryegrass addition gave an increased biochar350 mineralisation of 33% and 40%, and increased biochar700 at 137% and 70%, in the low and high pH soils, respectively. Certainly, on the basis of our results, if biochar is used to sequester carbon a priming effect may occur, increasing CO₂-C evolved from soil and decreasing soil organic C. However, this will be more than compensated for by the increased soil C caused by biochar incorporation. A similar conclusion holds for accelerated mineralisation of biochar due to incorporation of fresh labile substrates. We consider that our results are the first to unequivocally demonstrate the initiation, progress and termination of a true positive priming effect by biochar on native soil organic C.     

Interactive effects of warming, soil humidity and plant diversity on litter decomposition and microbial activity

Human activity has induced a multitude of global changes that are likely to affect the functioning of ecosystems. Although these changes act in concert, studies on interactive effects are scarce. Here, we conducted a laboratory microcosm experiment to explore the impacts of temperature (9, 12 and 15 °C), changes in soil humidity (moist, dry) and plant diversity (1, 4, 16 species) on soil microbial activity and litter decomposition.We found that changes in litter decomposition did not mirror impacts on microbial measures indicating that the duration of the experiment (22 weeks) may not have been sufficient to determine the full magnitude of global change effects. However and notably, changes in temperature, humidity and plant litter diversity/composition affected in a non-additive way the microbial parameters investigated. For instance, microbial metabolic efficiency increased with plant diversity in the high moisture treatment but remained unaffected in low moisture treatment suggesting that climate changes may mask beneficial effects of biodiversity on ecosystem functioning. Moreover, litter decomposition was unaffected by plant litter diversity/composition but increased with increasing temperature in the high moisture treatment, and decreased with increasing temperature in the low moisture treatment.We conclude that it is inevitable to perform complex experiments considering multiple global change agents in order to realistically predict future changes in ecosystem functioning. Non-additive interactions highlight the context-dependency of impacts of single global change agents.     

小麦–玉米轮作体系长期施肥对塿土微生物量碳、氮及酶活性的影响

以20年塿土小麦玉米轮作体系长期肥料定位试验为平台,探讨不同施肥模式下土壤化学肥力要素、微生物量碳氮及酶活性的响应。试验包括不施肥（CK）、单施氮肥（N）、氮磷（NP）、磷钾（PK）、氮磷钾（NPK）、NPK+秸秆（SNPK）以及不同量有机肥+NPK（M1NPK、M2NPK）等8种施肥模式。结果表明,与CK相比,长期施用NP提高土壤有机碳含量达34.0%、全氮34.0%、全磷58.5%、速效磷608.9%、微生物量碳23.3%、微生物量氮54.0%、蔗糖酶53.9%、脲酶132.6%、碱性磷酸酶29.9%以及脱氢酶40.9%。长期施用NPK与NP效果相似,钾素效果甚微。作物秸秆还田配合氮磷钾化肥与氮磷钾相比没有明显影响土壤有机碳、氮和磷水平,但是显著提高微生物量碳的含量（29.5%）、碱性磷酸酶（23.0%）和脱氢酶（26.9%）的活性。有机肥配合氮磷钾与其它施肥处理相比,显著提升土壤化学肥力要素、微生物量碳氮和酶活性,特别是引起了磷素的大量富集（速效磷含量大于150 mg/kg）。因此,塿土不施有机物情况下,氮磷配合可以提高土壤化学和生物肥力,作物秸秆还田配合氮磷钾化肥的培肥效果优于氮磷钾化肥配合,而合理的有机无机肥配合是塿土提升化学肥力和保证生物健康的最佳施肥模式。     

控释肥残膜对小麦各生育期土壤微生物和酶活性的影响

为了探明控释肥树脂残膜对土壤环境可能造成的影响,采用小麦池栽试验研究了控释肥树脂残膜对土壤有关微生物数量和酶活性的影响。结果表明,控释肥树脂残膜使土壤细菌和放线菌数量分别相对增加了19.01%～62.87%和17.03%～132.39%,土壤脲酶、转化酶、中性磷酸酶活性分别提高了17.39%～85.71%、31.77%～158.40%、35.14%～189.47%,但对过氧化氢酶活性无明显影响。施肥处理（施肥不施残膜和施肥施残膜处理）显著增加了土壤转化酶和中性磷酸酶活性;但对土壤细菌和放线菌数量、过氧化氢酶和脲酶活性没有显著影响。控释肥树脂残膜施用量在90～360g/m2 范围内,对土壤细菌和放线菌数量以及有关土壤酶活性没有产生不良影响。     

小叶锦鸡儿固沙群落土壤微生物生物活性的季节动态

为探讨科尔沁沙地小叶锦鸡儿固沙群落土壤微生物生物量和酶活性的季节动态规律,选取6,11,24年生人工群落和天然群落为对象,研究了不同土层深度微生物生物量(C、N、P)和脲酶、蛋白酶、蔗糖酶、磷酸单酯酶、脱氢酶和多酚氧化酶的活性随季节的变化特征。土壤按5层取样:0-10cm,10-20cm,20-30cm,30-40cm,40-50cm。结果表明,6～24年生小叶锦鸡人工固沙群落及天然群落土壤微生物生物量表现出明显的季节性变化,微生物生物量C和N为夏季>秋季>春季;微生物生物量P为夏季>秋季;6种土壤酶的活性也随季节发生显著变化,夏季显著高于春季和秋季。土壤微生物生物量和6种酶的活性随群落发育年限的增长而升高,但人工群落土壤的生物活性始终低于天然群落。6种土壤酶活性和微生物生物量最高值均出现在土壤表层(0-10cm),随着土层的加深生物活性逐渐降低。     

不同恢复方式下退化岩溶山区土壤微生物特性

为了探讨恢复方式对退化岩溶山区土壤生化特性的影响,对贵州南部岩溶山区退化地、不同恢复方式和对照原始林下土壤养分、微生物活性及群落结构进行了比较研究。结果表明:与退化地相比,不同恢复方式下土壤养分和微生物特性指标都有不同程度的提高;相对于对照原始林,不同恢复方式下大部分土壤养分和微生物功能活性指标的恢复程度分别为40%~75%和27%~60%,暗示着退化岩溶山区土壤微生物功能活性恢复滞后于土壤养分恢复;除土壤有效磷和蔗糖酶活性外,人工恢复方式(花椒和人还)的大部分土壤养分含量和土壤微生物活性恢复程度分别为对照的34%~47%和17%~47%,均低于自然恢复方式(还草和自还)的土壤养分和微生物活性恢复程度(对照的52%~78%和31%~61%),意味着自然恢复方式提高土壤质量效果优于人工恢复方式;不同恢复方式下植被状况影响着退化岩溶土壤细菌的分子群落结构,但不同恢复方式间土壤细菌种的丰富度和多样性差异均不明显。     

低分子量根系分泌物对土壤微生物活性及团聚体稳定性的影响

以黑土为供试土壤,采用化学试剂模拟根系分泌物进行室内培养试验,研究低分子量根系分泌物及主要组分对土壤微生物活性及团聚体稳定性的影响.结果显示,与对照相比,低分子量根系分泌物及主要组分短期内均显著提高了土壤微生物的活性,促进了>250μm大团聚体,尤其是>2 000μm团聚体的形成,显著增强了土壤团聚体的稳定性.低分子量根系分泌物中葡萄糖组分有利于土壤中细菌的生长,从而在细菌活动的驱动下,促进了微团聚体的形成;苹果酸和谷氨酸组分则有利于真菌的生长和繁殖,在真菌菌丝网络的缠绕和牯结作用下,促使微团聚体进一步胶结形成大团聚体.因此,低分子量根系分泌物中各主要组分共同参与并影响土壤的团聚化过程,改善根际微区土壤的结构,以利于植物根系的生长.     

磷灰石等改良剂对铜污染土壤的修复效果研究——对铜形态分布、土壤酶活性和微生物数量的影响

通过田间小区试验,研究了磷灰石、石灰、木炭、猪粪、铁粉对Cu污染土壤中Cu形态分布、土壤酶活性和微生物数量的影响。结果表明:改良剂的处理减少了离子交换态Cu的百分含量,增加了碳酸盐、铁锰氧化物、有机结合态Cu的百分含量,但对残留态Cu百分含量影响不大。磷灰石、石灰、木炭显著提高了土壤过氧化氢酶、酸性磷酸酶活性和土壤pH,但对脲酶活性影响较小。改良剂不同程度地增加了土壤细菌和真菌数量,且细菌和真菌数量均与土壤pH呈正相关关系,特别是真菌,与pH的相关系数达0.90。pH是影响Cu化学形态、土壤酶活性和微生物数量的主要因素,磷灰石、石灰和木炭显著提高了土壤pH,降低Cu的活性,增加了土壤酶活性和微生物数量,对Cu污染土壤具有较好的修复效果。     

Effects of inoculants on the fermentation characteristics and in vitro digestibility of reed canary grass (Phalaris arundinacea L.) silage on the Qinghai‐Tibetan Plateau

To effectively use local available grass resources to cover the winter feed shortage on the Qinghai‐Tibetan Plateau, direct‐cut and wilted reed canary grass (RCG) silages were prepared by using a rolled‐bale system, and their ensiling characteristics and in vitro digestibility were studied. Silages were treated without (control) or with inoculants including LP (Lactobacillus plantarum), LPLB (L. plantarum, L. buchneri), and LPLBc (L. plantarum, L. buchneri, and cellulase), and were stored at ambient temperature (5.7–14.6°C) for 90 days. Compared with control, the inoculated silages increased (p < .05) lactic acid and acetic acid contents, and reduced (p < .05) final pH value and ammonia‐N ratio of total N. The highest WSC content (41.2 g/kg DM) occurred for LPLB‐inoculated silage, whereas LPLBc‐treated silage displayed the lowest contents of NDF (522.9 g/kg DM) and ADF (275.5 g/kg DM). In addition, LPLBc‐inoculated silage had the highest in vitro gas production (51.0 ml/g DM), in vitro DM digestibility (619.3 g/kg DM), and metabolic energy (9.6 kJ/kg DM). These results confirmed that treatments with inoculants at ensiling could improve silage fermentation and in vitro digestibility of RCG, and this could be a potential winter feed for animals on the Qinghai‐Tibetan Plateau.