土壤微生物生物量氮及其在氮素循环中作用

简述了土壤微生物生物量氮的含量及其影响因素,阐述了其在土壤氮素循环中的重要作用,着重讨论了其与可矿化氮、矿质氮、有机氮和固定态铵之间的关系,指出土壤微生物生物量氮与供氮因子间的关系在氮素循环研究中有非常重要的作用,可为调控土壤氮素的供应状况,减少氮素损失,提高氮肥利用率提供科学依据,并提出了需要深入研究的问题。     

菜园土壤无机氮解吸特性对硝态氮流失潜能的影响

根据土壤氮素解吸模型,通过盆栽试验研究解吸特征参数对土壤渗漏水硝态氮浓度的影响.结果表明：土壤氮素可解吸量Q、土壤溶液氮初始浓度Cli和C₁/比值与土壤渗漏水硝态氮浓度呈非线性关系,在较低氮解吸特征值时则呈线性关系,由此提出“双速率转折点”概念评价土壤硝态氮流失潜能.当耕层土壤氮素解吸特征值超过“双速率转折点”X₀时,硝态氮浓度的增加速率将以非线性形式迅速提高,反之将稳定在较低水平.     

旱地小麦不同栽培条件对土壤硝态氮残留的影响

在陕西渭北旱塬进行了2a田间试验,研究不同栽培模式、施氮量和小麦种植密度对旱地硝态氮残留的影响。结果表明,种植小麦2a后0~200 cm土壤剖面中残留硝态氮58.6~283.9 kg/hm2,数量可观,短期内在渭北旱塬深厚的土壤中不会对地下水造成威胁,但夏季休闲期间容易下迁至作物无法吸收的土壤深度。与常规无覆盖模式相比,地膜覆盖和垄沟种植显著提高了作物对氮素的吸收,但同时也增加了土壤0~200 cm的硝态氮残留,这与地膜覆盖导致有机氮矿化增加有关;秸秆覆盖对作物氮素吸收和硝态氮残留均没有明显影响。施氮量低于120 kg/hm2时,各种栽培模式土壤剖面残留硝态氮的分布差异较小,只有地膜覆盖和垄沟种植处理在土壤表层有少量硝态氮累积;施氮量为240 kg/hm2时,无覆盖和秸秆覆盖土壤60~120 cm深度都有明显累积峰,地膜覆盖和垄沟种植土壤残留硝态氮则在60 cm以上土层累积较多。小麦种植密度也影响了各种栽培模式土壤硝态氮及其分布特点。垄沟种植条件下,从土壤表层到200 cm的深层,垄上土壤残留硝态氮均显著高于沟内土壤;上层差异最大,随着土壤深度的增加其差异逐渐降低;随着施氮量的增加,这种差异显著增大;随小麦种植密度的增加则显著降低。随着施氮量增加,小麦吸氮量和土壤中残留硝态氮量均显著提高;施氮增加的残留硝态氮占施氮量的0.3%~44.6%。垄沟种植模式施氮增加的残留硝态氮最多,地膜覆盖处理次之,垄沟种植处理垄上土壤增加量远远高于沟内土壤。施氮量提高1倍,增加的残留硝态氮量平均提高了3倍多。提高小麦种植密度,施氮增加的残留硝态氮平均减小13.2 kg/hm2。由于种植密度增加显著提高了小麦对氮素的吸收,因此硝态氮残留有降低的趋势。其中,秸秆覆盖模式80~140 cm土层降低显著;地膜覆盖条件下高密与低密残留硝态氮的差异主要在深层;垄沟模式中,低密度种植硝态氮残留量在整个土壤剖面都高于高密度处理;而无覆盖条件下,残留硝态氮则随种植密度的提高呈增加趋势。     

土壤微生物量氮含量、矿化特性及其供氮作用

论述了土壤中微生物体氮的含量及其影响因素,土壤微生物量氮的矿化特性及其与土壤矿化氮间的关系,土壤微生物量氮含量与土壤供氮指标间的关系等。提出研究不同生态系统中土壤微生物量氮的含量及其变化规律,不同耕作栽培措施对土壤微生物量氮含量的影响。土壤微生物量在土壤氮素保持和释放中的作用,土壤微生物量氮的转化率与其供氮量间的关系;土壤微生物量氮与作物氮素吸收间的关系等,是土壤微生物量氮方面应重点研究的问题。     

冬小麦生育期农田尺度下土壤硝态氮淋失动态的数值模拟

在北京通州区永乐店田间试验的基础上 ,假设土壤由一系列不发生相互作用的一维土柱组成 ,根据实测的土壤有机质含量 ,假定土壤有机氮的矿化作用速率常数 (零级动力学 )和有机质含量成正比 ,运用 HYDRUS- 1D软件 ,分别就考虑和不考虑土壤有机氮的矿化速率的空间变异性这两种方案 ,对 2 0 0 0～ 2 0 0 1年冬小麦生长条件下农田尺度土壤氮素转化和硝态氮淋失规律进行了数值分析。两种方案的模拟结果表明 :考虑和不考虑土壤有机氮矿化速率的空间变异性对剖面 2 5 0 cm埋深处硝态氮淋失量的影响很小 ,其差异主要在于前者对土壤氮素的矿化量、固持及反硝化量、作物吸氮量的影响更大 ,其空间变异性高于不考虑矿化速率时的结果。剖面 2 5 0 cm埋深处平均的土壤水渗透量和累积硝态氮淋失量分别为 2 .2 5 mm、0 .0 0 984 m g/cm2 ,变异系数大于 1.4 6 ,属于强变异性。对模拟结果进行地统计学分析 ,结果表明 :剖面 2 5 0 cm埋深处的土壤水渗透量和硝态氮淋失量的半方差函数为纯块金形式 ,在空间上表现为相互独立。考虑有机氮矿化速率空间变异性时的土壤氮素净转化量、吸氮量均可用球状模型描述 ,其变程与土壤有机质含量的变程接近 ,约为 4 .7m;而不考虑有机氮矿化速率空间变异性时的土壤氮素净转化量用线性无基台值     

不同作物对旱地农田残留硝态氮的利用差异

在黄土高原南部娄土上,通过2a田间试验研究了小麦和苜蓿对土壤中不同累积量的残留硝态氮的利用差异。研究包括0—3 m土壤残留硝态氮累积量(设N1、N2、N3、N4、N5和N6共6个水平,残留硝态氮量依次增加)和作物种类(冬小麦和苜蓿)2个因素,分别采用冬小麦-夏休闲-冬小麦和苜蓿连作种植方式。结果表明,不施用氮肥条件下,冬小麦-休闲-冬小麦轮作周期与苜蓿连作2a内,土壤残留硝态氮的消长有明显差异。在第1季小麦生长期间,小麦的氮素携出量(63.9—130.3 kg/hm2)、氮素携出量占播前残留硝态氮量的比例(18%—27%)及氮素携出量占该生长季硝态氮减少量的比例(29%—62%)均显著高于同期的苜蓿处理。在第2个生长季内,苜蓿的氮素携出量是小麦当季氮素携出量的近6倍,但由于苜蓿固氮作用强烈,至第2生长季结束后,0—3 m土壤硝态氮量与苜蓿播前相比平均只减少了72.4 kg/hm2,而麦田0—3 m土壤硝态氮量与小麦播前相比减少了158.3 kg/hm2。在短期内如果通过种植作物消耗土壤剖面的残留硝态氮,冬小麦比苜蓿更有优势。第1季小麦氮素携出量与小麦播前0—2 m(r=0.920**)和0—3 m(r=0.857*)土层残留硝态氮量呈显著或极显著正相关,与0—1 m土层残留硝态氮量没有显著相关性;第1生长季苜蓿氮素携出量与播前0—1 m土壤硝态氮累积量呈显著正相关关系(r=0.846*),而与0—2 m和0—3 m土壤硝态氮累积量的相关性并不显著。小麦比苜蓿能利用更深土层中的硝态氮。随着播前0—3 m土壤残留硝态氮的增加,小麦和苜蓿地上部氮素携出量呈增加的趋势,硝态氮表观损失也显著增加。     

间作对马铃薯种植土壤硝化作用和硝态氮供应的影响

土壤硝态氮供应对满足作物氮素需求至关重要,但间作如何影响土壤硝态氮供应及其作用机制尚不清楚。本研究基于4个氮水平(N₀, 0 kg·hm^-2; N₁, 62.5 kg·hm^-2; N₂, 125 kg·hm^-2; N₃, 187.5 kg·hm^-2)的马铃薯单作、马铃薯与玉米间作小区试验,分析土壤硝态氮含量与强度、硝化势和氨氧化功能基因丰度的差异,探讨间作影响土壤硝态氮供应和氮调控的机理。结果表明: 土壤硝态氮含量和强度随施氮量增加而升高,但同一施氮水平下间作均低于单作。施氮提高了土壤硝化势,且单作的响应高于间作。土壤中氨氧化细菌(AOB)的amoA基因丰度大于氨氧化古菌(AOA),二者在间作时均随施氮量增加呈现先增加后降低的趋势;相同施氮量下,间作的AOA和AOB基因丰度(除N₂外)均低于单作。相关分析、回归分析和主成分分析显示,马铃薯间作后,土壤AOB、AOA的amoA基因丰度下降,硝化势减弱,导致土壤硝态氮含量和强度降低。因此,间作导致土壤硝态氮供应降低与土壤氮转化的微生物过程有关,间作条件下的马铃薯种植应注意保障土壤氮素供应。     

菜园土壤无机氮解吸特性对硝态氮流失潜能的影响

根据土壤氮素解吸模型,通过盆栽试验研究解吸特征参数对土壤渗漏水硝态氮浓度的影响.结果表明：土壤氮素可解吸量Q、土壤溶液氮初始浓度Cli和C₁/比值与土壤渗漏水硝态氮浓度呈非线性关系,在较低氮解吸特征值时则呈线性关系,由此提出“双速率转折点”概念评价土壤硝态氮流失潜能.当耕层土壤氮素解吸特征值超过“双速率转折点”X₀时,硝态氮浓度的增加速率将以非线性形式迅速提高,反之将稳定在较低水平.     

华北地区夏玉米土壤硝态氮的时空动态与残留

为了进一步明确华北地区冬小麦-夏玉米种植体系周年氮肥利用效率及其影响因素与机制,在试验区夏玉米生育期年均降雨量400mm左右,轻壤质底粘潮土中等土壤肥力条件下,通过设计不同施氮量(0、90、180、270kgN/hm2)处理,重点研究了夏玉米大田土壤硝态氮动态与残留积累情况。试验结果表明,夏玉米根系生物量最大值出现在吐丝期,最大根系分布深度约为1.2m。根干重密度(g/m3)随土壤深度增加而明显降低。根群主要分布在表土层,0～80cm土体根重比例达95%以上,1m以下根重比例不足1%。土壤硝态氮测定表明,从播种前到收获期,各施氮量处理(0、90、180、270kgN/hm2)2m土体土壤硝态氮平均含量均表现出“N”型曲线变化趋势。在玉米收获期,施氮处理(90～270kgN/hm2)2m土体均有明显的硝态氮残留积累,并且残留积累量随着施氮量增加而增大,施氮处理下层土壤(120～200cm)硝态氮残留积累量比不施氮处理高出50.4～95.4kgNO3-N/hm2。这说明,在玉米生育期降水影响下氮肥发生了淋溶,有部分氮肥已经向下移出玉米根区以外,积累在下层土壤中。这些残留积累在下层土壤中的硝态氮对于玉米来说很难被吸收利用,不仅降低了氮肥的利用率,也成为污染地下水的潜在隐患。分析表明,各施氮处理籽粒产量和植株吸氮量都显著大于不施氮处理,但施氮处理之间比较,籽粒产量和植株吸氮量并无显著差异。90kgN/hm2、180kgN/hm2和270kgN/hm2施氮处理下,氮肥表观利用率分别为11.52%、13.37%、9.93%。根据本研究结果,从小麦-玉米种植体系考虑,玉米根区以下残留积累氮素的回收利用是提高周年氮肥利用率的一个重要方面,值得进一步研究。     

长期施肥及撂荒对土壤氮素矿化特性及外源硝态氮转化的影响

以黄土高原南部17年长期定位试验不同处理土壤为研究对象,研究了不同肥料处理及撂荒条件下土壤氮素矿化特性、灭菌与不灭菌条件下不同肥力土壤对施入外源硝态氮转化的影响.结果表明：氮磷钾化肥和有机肥配施（MNPK）及长期撂荒处理显著提高了土壤有机质和全氮含量以及土壤氮素矿化量和矿化率;氮磷钾化肥（NPK）处理虽然提高了土壤无机氮含量,但对土壤有机质、全氮、土壤氮素矿化量和矿化率的影响相对较小.高温高压灭菌显著增加了土壤铵态氮含量,但对不同处理土壤硝态氮含量无明显影响;在灭菌土壤培养过程中,土壤铵态氮含量呈显著增加趋势.同一土壤类型,不论灭菌与否,培养过程中施入土壤的硝态氮含量保持相对稳定,说明在本研究培养条件下,生物因素和非生物因素对外源硝态氮在土壤中的转化无明显影响.     

Factors affecting nitrogen dynamics in a semiarid woodland (Dry Chaco,Argentina)

In an effort to elucidate the factors affecting soil N dynamics in the Dry Chaco ecosystem, soil respiration and microbial biomass N were measured for one year underneath 5 vegetation types: a leguminous tree (Prosopis flexuosa DC), a non-leguminous tree (Aspidosperma quebracho-blanco Schlecht.), a non leguminous shrub (Larrea spp.), the open interspaces, and a pure grassland. Ammonifier and nitrifier densities and N content in litter were also measured in some cases. Results were compared with previously reported N mineralization rates and soil fertility.During the dry season microbial biomass N and net N mineralization were low, while accretion of easily mineralizable C occurred (estimated through soil respiration rates in lab under controlled temperature and moisture). With the onset of rain, microbial biomass N and N mineralization increased markedly, resulting in a decrease in easily mineralizable C. Throughout the wet season N mineralization varied with soil moisture while microbial biomass N remained consistently high. Mean values of immobilized N in this ecosystem were high (20–140 mg kg^–1), of about the same order of magnitude as accumulated net N mineralization (50–150 mg kg^–1 yr^–1). Microbial decay in the dry season, considered as a source of easily mineralizable N, accounted for only 40% of gross N mineralization increase at the beginning of the wet season. Ammonifier densities correlated significantly with soil moisture and N mineralization, but nitrifiers did not.The highest values of total N, N mineralization, inorganic N, microbial biomass N, nitrifier densities, N content in litter, total organic C and easily mineralizable C were found under Prosopis and the lowest values under shrubs and the interspaces. The main differences between tree species were in N mineralization at the beginning of the wet season, in total and inorganic N pools, and in nitrifier densities; all of which were significantly lower under Aspidosperma than under Prosopis.N mineralization in the pure grassland was very low despite high values of total N and C sources. Although N immobilized in microbial biomass was similarly high under Aspidosperma, Prosopis and the pure grassland, net N mineralization rates were quite different.     

Grass species and soil type effects on microbial biomass and activity

We evaluated plant versus soil type controls on microbial biomass and activity by comparing microbial biomass C, soil respiration, denitrification potential, potential net N mineralization and nitrification in different soils supporting four grass species, and by growing a group of 10 different grass species on the same soil, in two experiments respectively. In the first experiment, none of the microbial variables showed significant variation with grass species while all variables showed significant variation with soil type, likely due to variation in soil texture. In the second experiment, there were few significant differences in microbial biomass C among the 10 grasses but there were significant relationships between variation in microbial biomass C and potential net N mineralization (negative), soil respiration (positive) and denitrification (positive). There was no relationship between microbial biomass C and either plant yield or plant N concentration. The results suggest that 1) soil type is a more important controller of microbial biomass and activity than grass species, 2) that different grass species can create significant, but small and infrequent, differences in microbial biomass and activity in soil, and 3) that plant-induced variation in microbial biomass and activity is caused by variation in labile C input to soil.     

Mechanisms of plant species impacts on ecosystem nitrogen cycling

Plant species are hypothesized to impact ecosystem nitrogen cycling in two distinctly different ways. First, differences in nitrogen use efficiency can lead to positive feedbacks on the rate of nitrogen cycling. Alternatively, plant species can also control the inputs and losses of nitrogen from ecosystems. Our current understanding of litter decomposition shows that most nitrogen present within litter is not released during decomposition but incorporated into soil organic matter. This nitrogen retention is caused by an increase in the relative nitrogen content in decomposing litter and a much lower carbon‐to‐nitrogen ratio of soil organic matter. The long time lag between plant litter formation and the actual release of nitrogen from the litter results in a bottleneck, which prevents feedbacks of plant quality differences on nitrogen cycling. Instead, rates of gross nitrogen mineralization, which are often an order of magnitude higher than net mineralization, indicate that nitrogen cycling within ecosystems is dominated by a microbial nitrogen loop. Nitrogen is released from the soil organic matter and incorporated into microbial biomass. Upon their death, the nitrogen is again incorporated into the soil organic matter. However, this microbial nitrogen loop is driven by plant‐supplied carbon and provides a strong negative feedback through nitrogen cycling on plant productivity. Evidence supporting this hypothesis is strong for temperate grassland ecosystems. For other terrestrial ecosystems, such as forests, tropical and boreal regions, the data are much more limited. Thus, current evidence does not support the view that differences in the efficiency of plant nitrogen use lead to positive feedbacks. In contrast, soil microbes are the dominant factor structuring ecosystem nitrogen cycling. Soil microbes derive nitrogen from the decomposition of soil organic matter, but this microbial activity is driven by recent plant carbon inputs. Changes in plant carbon inputs, resulting from plant species shifts, lead to a negative feedback through microbial nitrogen immobilization. In contrast, there is abundant evidence that plant species impact nitrogen inputs and losses, such as: atmospheric deposition, fire‐induced losses, nitrogen leaching, and nitrogen fixation, which is driven by carbon supply from plants to nitrogen fixers. Additionally, plants can influence the activity and composition of soil microbial communities, which has the potential to lead to differences in nitrification, denitrification and trace nitrogen gas losses. Plant species also impact herbivore behaviour and thereby have the potential to lead to animal‐facilitated movement of nitrogen between ecosystems. Thus, current evidence supports the view that plant species can have large impacts on ecosystem nitrogen cycling. However, species impacts are not caused by differences in plant quantity and quality, but by plant species impacts on nitrogen inputs and losses.     

Microbes drive global soil nitrogen mineralization and availability

Soil net nitrogen mineralization rate (N_min), which is critical for soil nitrogen availability and plant growth, is thought to be primarily controlled by climate and soil physical and/or chemical properties. However, the role of microbes on regulating soil N_min has not been evaluated on the global scale. By compiling 1565 observational data points of potential net N_min from 198 published studies across terrestrial ecosystems, we found that N_min significantly increased with soil microbial biomass, total nitrogen, and mean annual precipitation, but decreased with soil pH. The variation of N_min was ascribed predominantly to soil microbial biomass on global and biome scales. Mean annual precipitation, soil pH, and total soil nitrogen significantly influenced N_min through soil microbes. The structural equation models (SEM) showed that soil substrates were the main factors controlling N_min when microbial biomass was excluded. Microbe became the primary driver when it was included in SEM analysis. SEM with soil microbial biomass improved the N_min prediction by 19% in comparison with that devoid of soil microbial biomass. The changes in N_min contributed the most to global soil NH₄⁺‐N variations in contrast to climate and soil properties. This study reveals the complex interactions of climate, soil properties, and microbes on N_min and highlights the importance of soil microbial biomass in determining N_min and nitrogen availability across the globe. The findings necessitate accurate representation of microbes in Earth system models to better predict nitrogen cycle under global change.     

川西亚高山不同海拔森林土壤活性氮库及净氮矿化的季节动态

研究了川西理县毕棚沟不同海拔梯度(3600 m、3300 m和3000 m)森林群落土壤活性氮库及土壤净氮矿化速率的季节动态.结果表明: 研究区森林土壤活性氮库(铵态氮、硝态氮、微生物生物量氮和可溶性有机氮)及净氮矿化速率存在明显的季节变化,但不同形态土壤活性氮库的季节动态有一定差异.4个采样时期(非生长季与生长季初期、中期及末期)各海拔土壤硝态氮浓度(8.38～89.60 mg·kg^-1)均显著高于铵态氮浓度(0.44～8.43 mg·kg^-1).生长季初期各海拔梯度的土壤净氮矿化速率均表现为负值(-0.77～-0.56 mg·kg^-1·d^-1),而非生长季、生长季中期和末期均为正值.除硝态氮外,不同海拔的土壤铵态氮、微生物生物量氮和可溶性有机氮浓度的差异极显著,海拔对它们的影响与季节变化有关.该区土壤净氮矿化以硝化为主,且氮矿化过程不受海拔梯度的影响.冬季土壤净氮矿化明显(0.42～099 mg·kg^-1·d^-1),早春高的土壤无机氮可能为植物生长提供基础养分,也可能通过淋溶方式从系统中丢失.     

秸秆生物有机肥的施用对土壤供氮能力的影响

通过盆栽试验研究了秸秆生物有机肥的施用对土壤供氮能力的影响。结果表明,与原始秸秆相比,秸秆生物有以与尿素配合施用能明显提高土壤生物量碳,氮的含量,增幅约为3－4倍;土壤微生物量氮与土壤的供氮能力关系非常密切,前期发生强烈的微生物增殖过程,则中后期必会发生无机氮释放过程,这对作物的生长发育极为重要。同时提高了肥料氮利用率,揭示了秸秆生物有机肥配施无机氮肥改善土壤供氮能力的机理。     

中国森林生态系统氮循环特征与生产力间的相互关系

为了更好地了解森林生态系统净初级生产力(NPP)与氮循环之间的关系,本文对中国主要森林生态系统类型中净初级生产力(NPP)、枯落物氮、植被年氮积累量和土壤氮矿化速率之间的关系进行了研究分析.结果表明,我国森林生态系统净初级生产力与枯落物氮、植被年氮积累量和土壤氮矿化速率之间均存在比较显著的相关关系.其中相关性最显著的是净初级生产力与氮矿化速率之间的相关关系(R₂=0.7,n=37),其次是净初级生产力与植被年氮积累量之间的相关关系(R₂=0.60,n=37).     

Soil nitrogen dynamics as an indicator for longleaf pine restoration

Assessing the status of soil nutrients with their corresponding microbial communities provides important information about degraded soils during the restoration of coastal wet pine forests. Net nitrogen mineralization, nitrogen‐oxidizing bacteria (NOB), and soil microbial biomass were compared with patch‐derived volume along a 110‐year longleaf pine (Pinus palustris Mill.) chronosequence for identifying a trajectory and ecological benchmark during forest restoration. Net nitrogen mineralization rates decreased significantly in the maturing‐aged, pine patches, driven by a larger drop in net nitrification. Net nitrification and abundance of NOB were higher in young pine patches compared to soils from the maturing (86–110 years) pine patches. Gross nitrate fluxes followed the nonfungal portion of the soil microbial biomass along the chronosequence, declining in 64‐year‐old pine patches. Microbial biomass peaked in patches 17–34 years of age, but significantly declined in the older patches. Fungal biomass leveled off without decline. Ammonium was the major source of nitrogen within the maturing pine patches as well as the wetland patches, indicating that ammonium maintains longleaf pine during growth‐limiting conditions. Nitrate dominated during rapid tree growth, optimally in mesic conditions. The relative amounts of available ammonium to nitrate can be used to model nitrogen cycling in facultative‐wetland pine forests of the coastal United States as soils alternate between wet and mesic conditions. A key restoration benchmark occurred after 86 years of pine development when pine patch growth rates slowed, with lower numbers of NOB, when the nonfungal biomass leveled off, and net nitrification rates are at a minimum, during pine maturation.     

有机物料在维持土壤微生物体氮库中的作用

采用室内和田间培养试验,研究了有机物料矿化过程中土壤微生物体氮的变化,测定结果表明,有机物料对矿化过程和微生物体氮的影响,既与有机物料本身性质和组成有关,也与土壤肥力水平和施氮与否有关。加入Ｃ／Ｎ比高的有机物料后,微生物对矿质氮的净固定持续时间长,而加入Ｃ／Ｎ比小的则固定时间短;高肥力土壤上的固定时间比低肥力土壤短。不同有机物料对土壤微生物体氮的影响不同。从加绿豆茎叶、小麦茎叶、未腐解马粪、腐熟马粪、腐熟猪粪到厩肥,土壤微生物体氮依次减小,提供的有效能源物质丰富（如绿豆茎叶）或Ｃ／Ｎ比较高（如小麦茎叶）时影响效果突出。土壤肥力不同,有机物料对微生物体的影响效果不同,在低肥力土壤的效果突出,约为高肥力土壤的４倍,因此,在评价有机物料对土壤微生物体氮的影响时,既考虑有有机物料的性质和组成,也考虑土壤力水平、矿质氮含量和培养时期。