Effects of incubation temperatures on soil extracellular enzyme activity across different soil pH conditions

doi:10.1093/jpe/rtag102

J Plant Ecol ›› Advance articles DOI:10.1093/jpe/rtag102

Effects of incubation temperatures on soil extracellular enzyme activity across different soil pH conditions

Panpan Liu^1,2, Jiacong Zhou², Daryl L. Moorhead³, Megharaj Mallavarapu^4,5, Andrew J. Margenot^6,7, Taiki Mori^8,9, Xuyang Wang^10,11,12, Nicolas Fanin¹³, Raul Ochoa-Hueso^14,15, Dong Wang¹⁶, Zhao Jin², Ji Liu^2,17, Guopeng Liang^18,19, Yongxing Cui^20,21, Yang Yang², Jinzhao Liu², Yangyang Li², Xiao Chen²², Xiaojun Shi^1*, Ji Chen^2,23,24*

1 College of Resources and Environment, Southwest University, Chongqing 400716, China;
2 State Key Laboratory of Loess Science, Institute of Earth Environment, Chinese Academy of Sciences, Xi'an, China;
3 Department of Environmental Sciences, University of Toledo, 2801 W Bancroft St, Toledo, OH, 43606 USA;
4 Global Centre for Environmental Remediation (GCER), School of Environmental and Life Sciences, College of Engineering, Science and Environment, University of Newcastle, ATC Building, University Drive, Callaghan, NSW, 2308, Australia;
5 Cooperative Research Centre for Contamination Assessment and Remediation of the Environment (crcCARE), University of Newcastle, ATC Building, University Drive, Callaghan, NSW, 2308, Australia;
6 Department of Crop Sciences, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA;
7 Agroecosystem Sustainability Center, Institute for Sustainability, Energy, and Environment, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA;
8 Kyushu Research Center, Forestry and Forest Products Research Institute, FFPRI, Kurokami 4-11-16, Kumamoto 860-0862, Japan;
9 Department of Forest Site Environment, Forestry and Forest Products Research Institute, FFPRI, Matsunosato 1, Tsukuba, Ibaraki, 305-8687, Japan;
10 State Key Laboratory of Ecological Safety and Sustainable Development in Arid Lands, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Lanzhou 730000, China;
11 University of Chinese Academy of Sciences, Beijing 100049, China;
12 Naiman Desertification Research Station, Northwest Institute of Eco-Environment and Resources, Chinese Academy of Sciences, Tongliao 028300, China;
13 INRAE, Bordeaux Sciences Agro, ISPA, 33140, Villenave d'Ornon, France;
14 Department of Biology, IVAGRO, University of Cádiz, Cádiz, Spain;
15 Department of Terrestrial Ecology, Netherlands Institute of Ecology (NIOO- KNAW), Wageningen, The Netherlands;
16 International Joint Research Laboratory of Global Change Ecology, School of Life Sciences, Henan University, Kaifeng 475004, China;
17 Hubei Province Key Laboratory for Geographical Process Analysis and Simulation, Central China Normal University, Wuhan, China;
18 Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT, USA;
19 Institute for Global Change Biology, University of Michigan, Ann Arbor, MI, USA;
20 Sino-French Institute for Earth System Science, College of Urban and Environmental Sciences, Peking University, Beijing, China;
21 Institute of Biology, Freie Universität Berlin, Berlin 14195, Germany;
22 State Key Laboratory of Atmospheric Boundary Layer Physics and Atmospheric Chemistry;
(LAPC), Institute of Atmospheric Physics, Chinese Academy of Sciences, No. 40, Huayanli, Chaoyang District, Beijing, 100029 China;
23 Institute of Global Environmental Change, Department of Earth and Environmental Science, School of Human Settlements and Civil Engineering, Xi'an Jiaotong University, Xi'an, China;
24 Guanzhong Plain Ecological Environment Change and Comprehensive Treatment National Observation and Research Station, Xi'an, China
^*Corresponding author: Xiaojun Shi, shixj@swu.edu.cn.
Ji Chen, chenji@ieecas.cn.

Received:2026-01-12 Revised:2026-04-09 Accepted:2026-04-30 Published:2026-05-28
Supported by:
This work was supported by the National Natural Science Foundation of China (Grant No. 32471685, 32301405) and the Shaanxi Province Natural Science Foundation for Distinguished Young Scholar (Grant No. 2024JC-JCQN-32).

不同pH条件下培养温度对土壤胞外酶活性的影响

Abstract

Abstract: Soil extracellular enzymes are essential regulators of biogeochemical cycles, yet the conditions under which their activities are assayed, particularly incubation temperature, remain poorly standardized. This variation among studies can introduce substantial bias and complicate cross- study comparisons. Here, we examined hydrolase and oxidase activities across different soil pH conditions, with samples separately incubated at 10, 20, and 30 °C in acidic, neutral, and alkaline forest soils. Our results showed that hydrolase activities increased with increasing incubation temperature, though the pattern differed among soil pH conditions. In acidic soil, hydrolase activities were lower at 10 °C compared to 20 and 30 °C, with little difference between 20 and 30 °C. In contrast, neutral and alkaline soils showed a continuous increase in hydrolase activities from 10 to 30 °C. When data from all soils were combined, oxidase activities displayed no overall temperature effect. However, distinct pH-specific trends emerged: in acidic and neutral soils, oxidase activities increased with incubation temperature, whereas in alkaline soils it declined at higher temperatures. Across all incubation temperatures, soil pH was the primary environmental driver of both hydrolase and oxidase activities, whereas incubation temperature itself exerted no significant influence. This strong soil pH dependence masked the temperature effect, highlighting that pre-existing soil conditions fundamentally influenced the enzymatic thermal response. We propose a decision-making framework that selects incubation temperatures based on soil pH and enzyme class to improve the accuracy and cross-study comparability of measurements of soil extracellular enzyme activities.

Key words: Incubation temperatures, Hydrolases, Oxidases, Soil pH, Forest ecosystem, Decision framework

摘要：
土壤胞外酶是生物地球化学循环的关键驱动因子，然而其活性测定条件（尤其是培养温度）目前仍缺乏统一的标准。方法学上的差异不仅会引入较大的实验误差，也会给不同研究间的比较带来了困难。为此，本研究选取酸性、中性和碱性森林土壤为研究对象，分别在10、 20和30 ℃三个温度下进行培养，测定并比较了不同培养温度下土壤水解酶和氧化酶的活性。结果表明，土壤水解酶活性整体随培养温度的升高而增加，但其响应模式因土壤pH的不同而异：在酸性土壤中， 10 ℃下的水解酶活性显著低于20 ℃和30 ℃，而20 ℃与30 ℃之间差异很小；相比之下，在中性和碱性土壤中，水解酶活性在10~30 ℃范围内呈持续上升趋势。综合所有土壤样品的数据，培养温度对氧化酶活性无整体显著影响；然而，氧化酶活性对温度的响应有明显的pH特异性：在酸性和中性土壤中，氧化酶活性随培养温度的升高而增加，但在碱性土壤中，高温反而降低了其活性。在所有培养温度下，土壤pH始终是决定水解酶和氧化酶活性的主要环境影响因子，而培养温度本身的直接影响并不显著。这种强烈的pH依赖性掩盖了温度效应，表明土壤的本底条件从根本上制约着酶活性的温度响应特征。基于上述结果，本研究提出了一个依据土壤pH和酶类别来选择培养温度的决策框架，旨在提高土壤胞外酶活性测定的准确性以及不同研究间的可比性。

关键词: 培养温度, 水解酶, 氧化酶, 土壤pH, 森林生态系统, 决策框架

Panpan Liu, Jiacong Zhou, Daryl L. Moorhead, Megharaj Mallavarapu, Andrew J. Margenot, Taiki Mori, Xuyang Wang, Nicolas Fanin, Raul Ochoa-Hueso, Dong Wang, Zhao Jin, Ji Liu, Guopeng Liang, Yongxing Cui, Yang Yang, Jinzhao Liu, Yangyang Li, Xiao Chen, Xiaojun Shi, Ji Chen. Effects of incubation temperatures on soil extracellular enzyme activity across different soil pH conditions[J]. J Plant Ecol, DOI: 10.1093/jpe/rtag102.

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