Advance in Approaches of Determining Form and Mineralization Rate of Soil Organic Phosphorus
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摘要: 大多数土壤的有机磷（Po）占全磷30% ~ 65%，矿化后可被植物直接利用，成为生态系统生物有效磷重要来源。准确测定Po形态组成及矿化速率是科学利用土壤Po库的基础。本文目的是对目前常见土壤Po形态组成和矿化速率测定技术的优缺点、适用范围进行总结分析，为研究人员选择合适测定技术提供依据。搜集并分析近20年来中国知网和Web of Science数据库中92篇关于Po形态组成与矿化速率方法研究论文。介绍了3种常用Po形态测定技术（连续分级法、酶水解法和核磁共振法）和近年来兴起的3种Po矿化速率测定技术（同位素稀释技术、连续观测－差减法和模型法）的基本原理，分析了几种技术适用范围、主要优缺点和互补性。尽管目前仍无针对土壤Po形态组成和矿化速率“最佳”测定方法，但是，研究人员可根据待测土壤理化性质和实验目的选择“合适”的测定技术。提出未来应在酶水解技术标准化、13C和31P核磁共振技术联用、发展包含植物因素Po矿化速率测定技术、联用形态和矿化速率测定技术等方面开展深入研究。
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关键词:
- 土壤有效磷 /
- 有机磷矿化 /
- 植酸磷 /
- Hedley连续分级法 /
- 31P核磁共振波谱法 /
- 生物化学矿化 /
- 磷酸酶
Abstract: As an important component of total soil phosphorus (P), mineralized organic P (Po) is a major source of replenishing bioavailable P pool in many terrestrial ecosystems. There are many forms of Po with different bioavailability in soil. These forms of Po also differ in their ability of resisting the hydrolysis catalyzed by phosphatases. It is therefore necessary to accurately determine the forms and mineralization rates of soil Po to understand the biogeochemical cycling of Po and to use the legacy Po in soil. The aims were to clarify the advantages, disadvantages, scopes of application of several most commonly used approaches of determining the forms and mineralization rate of soil Po. The selected 92 papers in the databases of CNKI and the Web of Science in recent 20 years with a topic of the forms and mineralization rate of soil Po were discussed. Three approaches of determining the forms of soil Po were reviewed, including sequential fractionation procedure, enzyme addition assays and 31P nuclear magnetic resonance spectroscopy. There is no a ‘best’ approach of determining the forms and mineralization rate of soil Po. However, scientists can select a ‘proper’ one according to the physical-chemical properties of targeted soils and experimental goals. It is recommended to carry out more researches on the following fields: developing a standard procedure of enzyme addition assays, jointly utilizing 13C and 31P nuclear magnetic resonance spectroscopy to identify more unsolved Po forms, modifying methods of determining mineralization rate of soil Po to reflect the effects of plants, and combinedly utilizing approaches of determining the forms and mineralization rate to understand the mechanisms of transformation and mineralization of soil Po. -
表 1 土壤有机P形态测定主要方法优缺点比较
Table 1. Comparison of advantages and disadvantages of main methods for determining soil organic P forms
方法
Method原理
Principle优点
Advantage缺点
Disadvantage适用范围
Application scope连续分级法 溶P能力不同的浸提剂对P形态进行区分，依次采用中性、弱酸性、中酸性、强酸性提取剂对土壤无机P不同形态进行提取，与全P的差值即为Po含量。 可获得不同形态Po总量；能有效反映土壤Po对有效P库的补充潜力；适用性强、费用低廉。 仅“操作性”地划分P形态；无法获取Po的化学形态；步骤繁琐、耗时较长。 适用于各种土壤；可用于评估Po的有效性。 酶水解法 利用磷酸酶对磷酸酯类化合物催化后可得到正磷酸盐，结合磷酸酶底物的具体特征以及无机P的具体数量，从而推断样品中Po的含量与形态组成。 具有较为明确的生物化学机理；能很好地反映土壤Po的生物有效性和水解特性；能表征土壤Po在酶作用下释放有效P的潜力；专一性强、高效、可批量测定。 只能对Po形态进行大类划分（如类简单单酯磷、类双酯磷）；酶易失活、且用量多、价格高、成本偏高。 适合于不稳定态Po的测定；可用于评估Po的有效性；结合土壤理化性质，有助于制定调控Po形态转化的措施。 液相31P NMR 31P原子核的自旋量子数为半整数，在外加磁场条件下可以产生核磁共振现象。不同形态Po的原子核因屏蔽效应不同而出现不同的化学位移值，因此可通过分析波谱图上的化学位移值来辨识具有不同空间结构的有机或无机P化合物。 图谱丰富、能全面地从分子水平上表征土壤Po的形态组成；能同时检测多种Po；操作简单、需样量少、样品检测灵敏度高、精确度高。 碱性浸提液会导致部分Po降解，从而低估土壤中双酯磷的含量；耗时长，成本高；不适宜大规模常规分析。 适用于土壤样品中的各种P形态的定性和定量鉴定；较适宜的Po浓度范围：大于30 μg g–1干土。 固相31P NMR 同液相31P NMR。 不需要复杂的预处理和提取，可直接测定。 Po浓度较低土壤测定耗时长；灵敏度和光谱信噪比低。 顺磁物质含量较低的土壤；适用的Po浓度范围：大于100 μg g–1干土。 
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表 2 土壤有机P矿化速率测定方法优缺点比较
Table 2. Comparison of advantages and disadvantages of determination methods for soil organic P mineralization rate
方法
Method优点
Advantage缺点
Disadvantage适用范围
Application scope同位素稀释法 成本低、耗时短；效率高、制样简单；灵敏度高、准确率高。 结果受培养条件和时间的影响较大；受有效P浓度和微生物量P影响会降低准确性；具有放射性损害、实验步骤繁琐。 适用于大多数土壤类型、基于IEK的同位素方法不适用于原位研究。 连续监测－差减法 操作简单、成本低廉。 实验时间较长；适用土壤类型较少。 适用于有机层及一些吸附能力极弱的土壤。可用于原位监测。 模型法 应用范围广；评价标准统一，具有客观性。 不同环境条件下的土壤须开展前期实验来建立同位素稀释法所测矿化速率与替代指标之间的定量关系。 适用于大多数土壤类型；可用于原位研究。
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