氮胁迫对水曲柳幼苗养分吸收、利用和生物量分配的影响

树木受到环境胁迫时发生形态和生理上的改变,以便获取对其生长发育限制最严重的资源．在东北林区凋落物因受温度影响分解速度和有机物矿质化过程缓慢,森林树木常受到氮营养胁迫．通过温室栽培试验,对氮胁迫下水曲柳幼苗生长的生理学和形态学指标进行了研究．结果表明,在氮胁迫下,水曲柳幼苗的净氮吸收速率和净磷吸收速率都会减少,但在生长前期氮利用效率和磷利用效率高于氮供给充足(8mmol·L^-1)和过量(16mmol·L^-1)时的氮利用效率和磷利用效率．当氮供给浓度不足时,叶重比减少,而特定叶面积和根重比增加．相对生长速率随氮供给浓度增加而增加．在氮胁迫下净同化速率下降,导致总生物量下降．在幼苗生长前期,水曲柳幼苗处于氮胁迫时根／茎比显著大于氮供给充足或过量时的根／茎比。而在生长后期。根／茎比没有显著差别．     

Calibration of quantitative real-time TaqMan PCR by correlation with hyphal biomass and ITS copies in mycelia of Piloderma croceum

DNA-based quantification methods such as real-time TaqMan PCR allow a rapid and highly sensitive species-specific quantification of isolated fungal DNA material, but most quantification systems are only able to measure relative amounts of biomass or biomass changes during different treatments. In this experiment, an already established DNA quantification system for the ectomycorrhizal fungus Piloderma croceum, based on the ITS region of ribosomal DNA, was calibrated to absolute biomass to obtain a direct correlation between mycelial biomass and isolated ITS copies. Thin layers of sterile mycelia were cultured on slides. The mycelial biomass was calculated from measurements of the total hyphal length using image analysis, followed by determination of the mycelial volume, and multiplied by the specific weight of hyphae obtained from literature data. Using the very same mycelium, the number of ITS copies was quantified by TaqMan PCR. The mean value of 1047 (+/- 185) copies per mm hypha results in possible data for a direct conversion: one billion (10 (9)) ITS copies corresponded to 0.79 mg hyphal dry weight. For the ribosomal ITS multi-copy genes, the number of ITS copies could be calculated to approx. 152 (+/- 26) copies per dikaryotic cell. These conversion data now allow determination of the mycelial biomass of Piloderma croceum using real-time TaqMan PCR, a prerequisite for competition experiments with Piloderma croceum.     

Biomass, morphology and nutrient contents of fine roots in four Norway spruce stands

Fine root systems may respond to soil chemical conditions, but contrasting results have been obtained from field studies in non-manipulated forests with distinct soil chemical properties. We investigated biomass, necromass, live/dead ratios, morphology and nutrient concentrations of fine roots (<2 mm) in four mature Norway spruce (Picea abies [L.] Karst.) stands of south-east Germany, encompassing variations in soil chemical properties and climate. All stands were established on acidic soils (pH (CaCl₂) range 2.8–3.8 in the humus layer), two of the four stands had molar ratios in soil solution below 1 and one of the four stands had received a liming treatment 22 years before the study. Soil cores down to 40 cm mineral soil depth were taken in autumn and separated into four fractions: humus layer, 0–10 cm, 10–20 cm and 20–40 cm. We found no indications of negative effects of N availability on fine root properties despite large variations in inorganic N seepage fluxes (4–34 kg N ha⁻¹ yr⁻¹), suggesting that the variation in N deposition between 17 and 26 kg N ha⁻¹ yr⁻¹ does not affect the fine root system of Norway spruce. Fine root biomass was largest in the humus layer and increased with the amount of organic matter stored in the humus layer, indicating that the vertical pattern of fine roots is largely affected by the thickness of this horizon. Only two stands showed significant differences in fine root biomass of the mineral soil which can be explained by differences in soil chemical conditions. The stand with the lowest total biomass had the lowest Ca/Al ratio of 0.1 in seepage, however, Al, Ca, Mg and K concentrations of fine roots were not different among the stands. The Ca/Al ratio in seepage might be a less reliable stress parameter because another stand also had Ca/Al ratios in seepage far below the critical value of 1.0 without any signs of fine root damages. Large differences in the live/dead ratio were positively correlated with the Mn concentration of live fine roots from the mineral soil. This relationship was attributed to faster decay of dead fine roots because Mn is known as an essential element of lignin degrading enzymes. It is questionable if the live/dead ratio can be used as a vitality parameter of fine roots since both longevity of fine roots and decay of root litter may affect this parameter. Morphological properties were different in the humus layer of one stand that was limed in 1983, indicating that a single lime dose of 3–4 Mg ha⁻¹ has a long-lasting effect on fine root architecture of Norway spruce. Almost no differences were found in morphological properties in the mineral soil among the stands, but vertical patterns were apparently different. Two stands with high base saturation in the subsoil showed a vertical decrease in specific root length and specific root tip density whereas the other two stands showed an opposite pattern or no effect. Our results suggest that proliferation of fine roots increased with decreasing base saturation in the subsoil of Norway spruce stands.     

氮肥对不同无机碳含量土壤二氧化碳释放的影响

施用氮肥会导致土壤pH值降低,其对不同无机碳含量的土壤二氧化碳(CO₂)释放的影响如何,尚不清楚.采用室内密闭培养试验研究了氮肥及其配施硝化抑制剂(DCD)对不同无机碳含量土壤pH、矿质态氮和CO₂释放的影响.结果表明:与不施氮肥相比,施用氮肥不同程度地降低了水稻土、砂姜黑土、塿土3类供试土壤的pH,提高了土壤碳累积释放量,49 d培养结束时,土壤碳累积释放量分别提高了39.4%、23.4%和71.8%;氮肥配施DCD后显著抑制了土壤硝化作用的进行,至培养结束时,3类供试土壤pH值均显著高于仅施氮肥处理,水稻土、砂姜黑土CO₂平均释放量与仅施氮肥相比无显著性差异,塿土CO₂平均释放量比纯施氮肥平均降低了12.5%.土壤无机碳能有效缓冲由氮肥施入而导致的土壤酸化,氮肥施入后石灰性土壤CO₂释放不仅来源于土壤有机碳的矿化,可能还有一部分来源于无机碳的溶解释放.我国不同地区间土壤无机碳含量各有差异,长期大量氮肥投入下土壤酸化和无机碳库消耗问题应引起足够的重视.     

Ammonium,nitrate and dissolved organic nitrogen in seepage water as affected by compost amendment to European beech,Norway spruce,and Scots pine forests

Fertilization of nutrient-depleted and degraded forest soils may be required to sustain utilization of forests. In some European countries, the application of composts may now be an alternative to the application of inorganic fertilizers because commercial compost production has increased and compost quality has been improved. There is, however, concern that compost amendments may cause increased leaching of nitrogen, trace metals and toxic organic compounds to groundwater. The objective of this study was to assess the risk of ammonium (NH₄ ⁺), nitrate (NO₃ ^–) and dissolved organic nitrogen (DON) leaching following a single compost application to silty and sandy soils in mature beech (Fagus sylvatica L.), pine (Pinus silvestris L.) and spruce (Picea abies Karst.) forests at Solling and Unterlüß in Lower Saxony, Germany. Mature compost from separately collected organic household waste was applied to the soil surface at a rate of 6.3 kg m^–2 in the summer of 1997 and changes in NH₄ ⁺, NO₃ ^– and DON concentrations in throughfall and soil water at 10 and 100 cm soil depths were determined for 32 months. The spruce forests had the highest N inputs by throughfall water and the highest N outputs in both the control and compost plots compared with the pine and beech forests. Overall, the differences in total N outputs at 100 cm soil depth between the control and compost plots ranged between 0.3 and 11.2 g N m^–2 for the entire 32-month period. The major leaching of these amounts occurred during the first 17 months after compost amendments, but there was no significant difference in total N outputs (–0.2 to 1.8 g N m^–2) between the control and compost plots during the remaining 15 months. Most of the mineral soils acted as a significant sink for NO₃ ^– and DON as shown by a reduction of their outputs from 10 to 100 cm depth. Based on these results, we conclude that application of mature compost with high inorganic N contents could diminish the groundwater quality in the first months after the amendments. A partial, moderate application of mature compost with low inorganic N content to nutrient depleted forest soils can minimize the risk of NO₃ ^– leaching.     

Influence of Norway spruce seedlings on the nutrient availability in mineral soil and forest floor material

A greenhouse experiment was performed to evaluate the effect of Norway spruce (Picea abies (L.) Karst.) seedlings on net nutrient availability in five different growing media containing F- or H-layer and mineral soil originating from a haplic podzol in northern Sweden. The initial total amounts of eight nutrient elements (N, K, P, Ca, Mg, Mn, Fe, Zn) and exchangeable amounts of the same elements were analyzed in pots with or without spruce seedlings. In the planted pots seedling nutrient uptake was also estimated. After 26 weeks, higher net nutrient availability with seedlings was found in 25 out of the 40 (62%) growing media and nutrient element combinations. A positive seedling effect on net nutrient availability might be explained by rhizodeposition stimulating the soil microorganism activity and accelerating the weathering of minerals or by seedling roots promoting the nutrient providing processes through changes in soil chemical and physical properties. Nitrogen availability was primarily affected by what part of the forest floor the growing medium contained although the positive response to seedling presence was apparent. The positive net availability response of P, Ca, Mg, Mn, Fe and Zn to seedling presence was on the other hand relatively strong. In the case of P, K, and Zn the growing medium composition (if the F- and H-layer was pure or mixed with mineral soil) was also an important factor for the estimated net availability. Pure F-and H-layer provided greater P- and K-availability while the availability of Zn increased when mineral soil was added. The influence of growing plants ought to be considered when soil samples are used for assessing the nutrient availability.     

Effects of growth medium, nutrients, water, and aeration on mycorrhization and biomass allocation of greenhouse-grown interior Douglas-fir seedlings

Commercial nursery practices usually fail to promote mycorrhization of interior Douglas-fir [Pseudotsuga menziesii (Mirb.) Franco var. glauca (Beissn.) Franco] seedlings in British Columbia, which may account for their poor performance following planting in the field. We tested the effects of four nursery cultivation factors (nitrogen fertilization, phosphorus fertilization, watering, and soil aeration) and field soil addition on mycorrhization, survival, growth, and biomass allocation of interior Douglas-fir seedlings in a series of greenhouse experiments. Where field soil was added to the growing medium, mycorrhization and root/shoot ratios were maximized at lower levels of mineral nutrient application and aeration. Where field soil was not added, mycorrhization was negligible across all fertilization and aeration treatments, but root/shoot ratio was maximized at lower levels of mineral nutrients and the highest level of aeration. Regardless of whether field soil was added, intermediate levels of soil water resulted in the best mycorrhizal colonization and root/shoot ratios. However, field soil addition reduced seedling mortality at the two lowest water levels. A cluster analysis placed ectomycorrhizal morphotypes into three groups (Mycelium radicis-atrovirens Melin, Wilcoxina, and mixed) based on their treatment response, with all but two morphotypes in the mixed group whose abundance was maximized under conditions common to advanced seedling establishment. For maximal mycorrhization and root development of interior Douglas-fir seedlings, nurseries should minimize addition of nitrogen and phosphorus nutrients, maximize aeration, provide water at moderate rates, and, where possible, add small amounts of field soil to the growing medium.     

Monitoring uptake and contents of Mg,Ca and K in Norway spruce as influenced by pH and Al,using microprobe analysis and stable isotope labelling

In a model system using intact spruce trees (Picea abies [L.] Karst.) we followed the path of magnesium, calcium and potassium during uptake into the root and during long-range transport into the shoot, by multiple stable isotope labelling. The roots of two- and three-year-old spruce trees originating from soil culture were removed from the soil and, in part or in toto, exposed to labelling solutions containing the stable isotopes ²⁵Mg or ²⁶Mg, ⁴¹K and ⁴²Ca or ⁴⁴Ca. Optical-emission-spectroscopy (ICP-OES) of plant fractions and labelling solutions was combined with the quantitative analysis of stable isotope ratios in sections of shock frozen, cryosubstituted material using the laser-microprobe-mass-analyser (LAMMA). This combination allowed us to distinguish, both in bulk samples and on the cellular level between (i) the fraction of elements originally present in the plant before the start of the labelling, (ii) the material taken up from the labelling solution into the plant and (iii) any material released by the plant into the labelling solution.In single-root labelling experiments, roots of three-year-old spruce trees, grown in nursery soil, were exposed to various pH conditions. The exchange of Mg and Ca with the labelling solution was nearly 100% in the cell walls of the mycorrhized finest roots. This exchange was only slightly affected by a step down to pH 3.5. The absolute Mg and Ca content in the cell walls was moderately reduced by incubation at pH 3.5 and strongly reduced in the presence of Al at this pH. After a pH 3.5 and 2 mM Al treatment we found Al in the xylem cell walls and the cortex cell lumina at elevated concentrations. To analyse the combined effect of high Al and high proton concentrations on the long-range transport, we used a split-root system. The root mass of an intact two-year-old spruce tree, grown in mineral soil, was divided into even parts and both halves incubated in solutions with two sets of different stable isotopes of Mg and Ca (side A: no Al, ²⁵Mg and ⁴²Ca; side B: +Al, ²⁶Mg and ⁴⁴Ca) and ⁴¹K on both sides. We observed a large uptake of Mg, Ca and K into the plant and a pronounced release. The net uptake of all three elements was lower from the Al-doted solution. In cross-sections of the apical shoot we found after seven-day labelling period about 60–70% of the Mg and Ca and 30% of the K content in the xylem cell walls originating from both labelling solutions. The clear majority of the Mg and Ca label originated from the Al-doted side.     

施用控释氮肥对稻田土壤微生物生物量碳、氮的影响

借助农业部望城红壤水稻土生态环境野外观测试验站的控释氮肥试验,研究了施用控释氮肥对水稻不同生育期间稻田土壤微生物生物量碳、氮动态变化的影响。试验共设5个处理:①CK,(不施氮肥);②Urea(施用尿素);③CRNF(施用与处理②等氮量的控释氮肥);④70%CRNF(施用控释氮肥,用氮量为处理②的70%);⑤50%CRNF+M(施用控释氮肥和猪粪,总氮量为处理②的70%,其中控释氮肥用量为处理②的50%,猪粪含氮量为处理②的20%)。结果表明,施肥后10 d,施氮处理土壤微生物生物量碳和氮均达最高,随生育进程推进逐渐下降,成熟期有一定的回升;施肥初期,施用等氮量的控释氮肥处理(CRNF)土壤微生物量碳、氮含量较尿素处理(Urea)分别增加5.4%和22.5%,而水稻生育中后期,控释氮肥处理(CRNF)土壤微生物量碳、氮含量较尿素处理(Urea)下降幅度大,该处理向地上部提供氮素营养较尿素处理高;施氮量较高的CRNF处理,土壤微生物生物量碳低于控释氮肥节氮处理(70%CRNF),但在大多数取样时期,土壤微生物量氮高于控释氮肥节氮处理(70%CRNF);控释氮肥配施有机肥的节氮处理较其他单施化肥处理显著增加土壤微生物生物量碳、氮含量。控释氮肥与有机肥配施,不仅能节约氮肥用量,而且能明显地提高土壤微生物生物量碳、氮的含量。     

Effect of exposure to fluoride,nitrogen compounds and SO2 on the numbers of spruce shoot aphids on Norway spruce seedlings

Summary Development of spruce shoot aphid (Cinara pilicornis Hartig) populations was monitored in natural and artificial infestations of Norway spruce (Picea abies Karst.) seedlings, exposed to air pollutants in an experimental field. The pollutants, applied both singly and in mixtures, were gaseous sulphur dioxide, NaF (30 mg l^-1 F) and Ca(NO₃)₂ or (NH₄)₂SO₄ in aqueous solutions (200 mg l^-1 N). Aphid numbers on 10 seedlings in each treatment and two control plots were counted at 2-week intervals. At the beginning of the experiment aphid numbers did not differ between treatments. Aphid populations peaked in late June and early July. All the pollutants and their combinations significantly increased the numbers of aphids per seedling. Four apterous females were transferred to spruce seedlings which were growing in containers in the same plots. After 4–5 weeks aphid numbers were significantly higher in the fluoride treatment and in the combined treatment of fluoride, nitrogen and SO₂. The pollution treatments did not have a significant effect on shoot growth. Concentrations of F and S in needles were higher in treatments involving these pollutants. There were no significant differences in the concentrations of free amino acids in shoot stems between control and fluoride treatment. However, the relatively low concentration of arginine in the F treatment at the end of the growing season might indicate disturbances in the nitrogen metabolism of spruce seedlings.     

Effect of initial fertilisation on biomass and nutrient contentof Norway spruce and Douglas-fir plantations at the same site

Norway spruce (NS) and Douglas-fir (DF) are among the main species used for production forestry in France. In low-elevation mountains and under-acidic conditions, they often occupy the same ecological situations. It is therefore of paramount interest to have a good understanding of how the two species behave under similar conditions and how they react to site improvement by fertilisation. The study stands are part of an experimental stand located in the estate forest of Breuil-Chenue in the Morvan (east central part of France). Its aim is to compare the impact of change in species on ecosystem functions. Destructive sampling of 10 trees per stand, distributed over the whole spectrum of inventoried classes of circumference at breast height (c _1.30), was carried out within four stands, e.g., fertilised and control (non-fertilised) NS; fertilised and control (non-fertilised) DF. Allometric relationships between c _1.30 and biomass or nutrient content per tree compartment were calculated. These equations were applied to the stand inventory for quantifying stand biomass and nutrient content on a hectare basis. The standard deviations of results were estimated using Monte-Carlo simulations. Specific emphasis was given to explain the origin of differences observed between species and treatments, i.e., changes in carbon allocation leading to specific allometric relationships, changes in stand structure (tree size distributions) and changes in stand density due to mortality.DF was more productive than NS (+28% for total tree biomass, +50% for ligneous biomass and +53% for stem wood). Both NS and DF were affected by fertilisation but in the case of NS, effects on the crown_c _1.30 relationship and on average tree growth were predominant while in the case of DF, the stem_c _1.30 relationship and stand density were affected by changes in soil fertility. The general fertilisation effect was an increment of 40% of ligneous dry matter for DF and only 22% for NS. In both cases, the amount of wood biomass produced per unit of leaf biomass (on a tree basis and, to a lesser extent, on a per hectare basis) was greater in fertilised plots. However, in the case of NS, the same amount of wood biomass was produced from a smaller quantity of leaves while in the case of DF, the same amount of leaves produced more wood biomass.The amount of nutrients in total ligneous biomass was higher for N, P and K, but lower for Ca and Mg, in DF than in NS. A high variability was observed between nutrient content of the different compartments, e.g., DF < NS for needles (except Mg), DF < NS for K, Ca and Mg for stem wood and DF > NS for N and P of stem wood. Fertilisation did not considerably change the hierarchy. On the basis of this study, all the indexes concerning stand production, wood density, nutrient use efficiency and response to fertilisation gave a net advantage to DF. This information is highly relevant for both ecological and practical purposes.     

Effects of mycorrhizal colonization on biomass production and nitrogen fixation of black locust (Robinia pseudoacacia) seedlings grown under elevated atmospheric carbon dioxide

Interactive effects of elevated atmospheric CO₂ and arbuscular mycorrhizal (AM) fungi on biomass production and N₂ fixation were investigated using black locust ( Robinia pseudoacacia ). Seedlings were grown in growth chambers maintained at either 350 μmol mol⁻¹ or 710 μmol mol⁻¹ CO₂. Seedlings were inoculated with Rhizobium spp. and were grown with or without AM fungi. The ¹⁵N isotope dilution method was used to determine N source partitioning between N₂ fixation and inorganic fertilizer uptake. Elevated atmospheric CO₂ significantly increased the percentage of fine roots that were colonized by AM fungi. Mycorrhizal seedlings grown under elevated CO₂ had the greatest overall plant biomass production, nodulation, N and P content, and root N absorption. Additionally, elevated CO₂ levels enhanced nodule and root mass production, as well as N₂ fixation rates, of non- mycorrhizal seedlings. However, the relative response of biomass production to CO₂ enrichment was greater in non-mycorrhizal seedlings than in mycorrhizal seedlings. This study provides strong evidence that arbuscular mycorrhizal fungi play an important role in the extent to which plant nutrition of symbiotic N₂-fixing tree species is affected by enriched atmospheric CO₂.     

Effect of wood ash application on the biomass distribution and physiological state of Norway spruce seedlings on sandy soils

The effect of wood ash applied as a fertiliser (0.25, 0.5 and 1 kg m^–2) to nutrient poor sandy soil on 4-year-old Norway spruce (Picea abies) growing in small-scale sample plots was studied. Analyses carried out with roots, stems, needles and shoots of different age showed that an increase in the pH level and K and Ca concentrations and a decrease in N and P concentrations in the soil was accompanied by a disbalance of nutrients in tree compartments. Stimulation of pigment synthesis in Norway spruce needles was observed, and no disbalances occurred in the pigment system (Chl a/Chl b, TChl/Car) guaranteeing normal process of photosynthesis. Biomass responses to wood ash application depended on the age of needles and shoots, being only slightly notable in current year organs. Inhibition of height growth of seedlings, but stimulation of root biomass was established.     

生境变化对栓皮栎幼苗营养元素含量的影响

以栓皮栎在我国天然分布的南界、中部和北界为研究样点,将不同纬度的栓皮栎实生幼苗移栽到同一地点,于生长旺季同时在种源地和移栽地进行取样和测定,探讨生境变化对栓皮栎幼苗生长旺季营养元素含量特征的影响.结果表明:生境变化对栓皮栎幼苗各器官N含量和茎P含量的影响显著,而对各器官K含量和叶片、根P含量影响较小;在种源地,栓皮栎各器官的N含量与纬度间呈极显著的正相关,茎、根的全P含量随纬度升高而降低,叶片N/P的变化不明显;移栽后,北界栓皮栎幼苗各器官的N、P含量显著降低,不同纬度来源对栓皮栎幼苗N、P含量的影响更为明显;各纬度栓皮栎叶片的N/P均有不同程度的升高.不同纬度栓皮栎幼苗不同器官的营养元素含量存在差异,其对生境变化的响应也不相同.     

喀斯特地区不同土地利用方式对土壤有机碳、全氮以及微生物生物量碳和氮的影响

以广西环江大才为代表,选择亚热带典型喀斯特峰林谷地样区,通过对样区土壤进行密集采样和测定分析,研究了土地利用方式对土壤有机碳（O_C）和全氮（TN）含量及土壤微生物生物量碳（B_C）和氮（B_N）含量的影响.结果表明,3种土地利用方式下,土壤有机碳含量在稻田和林地中基本相同,而旱地显著低于稻田和林地.土壤全氮含量为稻田显著高于林地,而林地显著高于旱地.土壤微生物生物量碳含量为稻田显著高于林地,林地显著高于旱地.土壤微生物生物量氮含量在稻田和林地中基本相同,而旱地显著低于稻田和林地.旱地土壤pH值显著低于稻田和林地土壤.3种土地利用方式下,土壤微生物生物量碳与土壤有机碳、土壤微生物生物量氮与全氮含量之间均呈显著的正相关关系.土壤微生物生物量碳和氮含量可以作为评价喀斯特地区土壤质量和肥力的指标之一,对土地利用方式响应较为敏感.     

The nitrogen mineralization rate of legume residues in soil as influenced by their polyphenol,lignin, and nitrogen contents

A 12-week greenhouse experiment was conducted to determine the effect of the polyphenol, lignin and N contents of six legumes on their N mineralization rate in soil and to compare estimates of legume-N release by the difference and ¹⁵N-recovery methods. Mature tops of alfalfa (Medicago sativa L.), round leaf cassia (Cassia rotundifolia Pers., var. Wynn), leucaena (Leucaena leucocephala Lam., deWit), Fitzroy stylo (Stylosanthes scabra Vog., var Fitzroy), snail medic (Medicago scutellata L.), and vigna (Vigna trilobata L., var verde) were incorporated in soil at the rate of 100 mg legume N kg^-1 soil. The medic and vigna were labeled with ¹⁵N. Sorghum-sudan hybrid (Sorghum bicolor, L. Moench) was used as the test crop. A non-amended treatment was used as a control. Net N mineralization after 12 weeks ranged from 11% of added N with cassia to 47% of added N for alfalfa. With the two legumes that contained less than 20 g kg^-1 of N, stylo and cassia, there was net N immobilization for the first 6 weeks of the experiment. The legume (lignin + polyphenol):N ratio was significantly correlated with N mineralization at all sampling dates at the 0.05 level and at the 0.01 level at 6 weeks (r²=0.866). Legume N, lignin, or polyphenol concentrations or the lignin:N ratio were not significantly correlated with N mineralization at any time. The polyphenol:N ratio was only significantly correlated with N mineralization after 9 weeks (r²=0.692). The (lignin + polyphenol):N ratio appears to be a good predictor of N mineralization rates of incorporated legumes, but the method for analyzing plant polyphenol needs to be standardized. Estimates of legume-N mineralization by the difference and ¹⁵N recovery methods were significantly different at all sampling dates for both ¹⁵N-labeled legumes. After 12 weeks, estimates of legume-N mineralization averaged 20% more with the difference method than with the ¹⁵N recovery method. This finding suggests that estimates of legume N available to subsequent crops should not be based solely on results from ¹⁵N recovery experiments.