Scientia Agricultura Sinica ›› 2020, Vol. 53 ›› Issue (13): 2678-2690.doi: 10.3864/j.issn.0578-1752.2020.13.016

• TECHNOLOGY AND MECHANISM FOR RECOVERY OF ABANDONED CROPLAND • Previous Articles     Next Articles

Responses of Soil Organic Carbon Fractionation and Microbial Community to Nitrogen and Water Addition in Artificial Grassland

XU Meng1,XU LiJun2,CHENG ShuLan3,FANG HuaJun1,3(),LU MingZhu1,YU GuangXia3,YANG Yan1,GENG Jing1,CAO ZiCheng3,LI YuNa3   

  1. 1Key Laboratory of Ecosystem Network Observation and Modeling, Institute of Geographic Sciences and Natural Resources Research, Chinese Academy of Sciences, Beijing 100101
    2Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences/Hulunber Grassland Ecosystem Observation and Research Station, Beijing 100081
    3College of Resources and Environment, University of Chinese Academy of Sciences, Beijing 100049
  • Received:2019-09-13 Accepted:2019-12-08 Online:2020-07-01 Published:2020-07-16
  • Contact: HuaJun FANG E-mail:fanghj@igsnrr.ac.cn

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Abstract:

【Objective】 Establishment of artificial grassland provides one solution for releasing some of the grazing pressure on natural grassland, and thus is conducive to the restoration of degraded grassland. The present study aimed to investigate the effects of nitrogen (N) addition and water supplement on soil organic carbon (SOC) fractionation, composition and activity of soil microbial community on artificial grassland established in Hulunber region, so as to provide insight into the effect of management practices on SOC sequestration and stability of artificial grassland. 【Method Experimental plots were designed with two factors of N addition (0, and 150 kg N·hm-2·a-1) and water addition (0, and 60 mm), and were constructed under three types of artificial grasslands, i.e., alfalfa monoculture, smooth bromegrass monoculture, and the mixed cultivation of alfalfa and smooth bromegrass. Soil samples were collected from experimental plots and SOC of bulk soil was fractionated into different particulate-size fractions by wet-sieving method. The abundance, composition and activity of soil microbial community were measured by phospholipid fatty acids (PLFA) and extracellular enzyme activity. Then, the effects of N addition and watering treatments on SOC fractions and soil microbial community, and investigated the relationship between these two components were analyzed. 【Result】 Three years of N fertilization and water addition exerted significant impact on SOC fractionation. Addition of N generally increased the content of particulate organic carbon (POC) in soils of alfalfa monoculture and the mixed cultivation but decreased the content of mineral associated organic carbon (MAOC), while water addition significantly increased the content of coarse POC in soils of smooth bromegrass monoculture. The activities of four soil extracellular enzymes but not the abundance or composition of soil microbial community were significantly affected by N and water addition. Nitrogen application without water addition significantly inhibited the activity of β-N-acetylglucosidase (NAG) in soils of alfalfa monoculture, but promoted that in soils of smooth bromegrass monoculture. The activities of cellobiohydorlase (CB) and NAG were significantly decreased by water addition in soils of two monoculture grasslands. Nitrogen application with water addition significantly inhibited the activities of β-glucosidase (βG), CB and NAG in soils of alfalfa monoculture, but significantly promoted CB activity in soils of the mixed cultivation. The changes in PLFA concentration of total community and specific microbial groups were positively correlated with changes in content of POC, but negatively correlated with MAOC. There were negative correlations between the activities of βG and CB, as well as the ratio of enzyme C/N and C/P with changes in POC content, and these correlations were stronger under treatments with water addition. 【Conclusion】 Results of the present study showed that in artificial grassland established in semi-arid Hulunbeier region, the addition of N fertilization significantly promoted the accumulation of labile SOC fraction, but decreased the proportion of recalcitrant fraction, which reduced the stability of soil carbon sequestration. Nitrogen fertilization and water supplement induced significant changes in activity of soil microbial communities, and these changes in enzyme stoichiometric ratios were closely related to changes in SOC fractions. These results implied that the differential demands for C, N and P by microbial communities were crucial in regulating the turnover of labile SOC fraction in artificial grassland.

Key words: artificial grassland, nitrogen addition, water addition, soil organic carbon, soil microbial community, extracellular enzyme activity, enzyme stoichiometric ratio, Hulunber

Table 1

Soil extracellular enzymes and corresponding substrates"

酶 Enzyme 缩写 Abbreviation 底物 Substrate
β-葡萄糖苷酶β-glucosidase βG 4-MUB-β-D-glucoside
纤维二糖水解酶 Cellobiohydorlase CB 4-MUB-β-D-cellobioside
β-N-乙酰氨基葡萄糖苷酶β-N-acetylglucosidase NAG 4-MUB-N-acetyl-β-D-glucosaminide
酸性磷酸酶 Acid phosphatase AP 4-MUB-phosphate

Table 2

Effect of N and water treatment on soil general properties under three forage cultivation types"

处理
Treatment		 pH 有机碳
SOC
(g·kg-1)		 总氮
TN
(g·kg-1)		 硝态氮
NO3--N
(mg·kg-1)		 铵态氮
NH4+-N (mg·kg-1)		 总可溶性氮
TDN
(mg kg-1)		 土壤水分
Soil moisture
(%)
苜蓿单播 Alfalfa monoculture
CK 7.0±0.4a 21.6±1.5a 2.1±0.1a 27.0±2.6c 4.6±0.4a 51.3±1.2c 12.4±0.3a
N 7.1±0.4a 20.2±0.7a 2.0±0.1a 52.9±6.9b 6.1±0.8a 78.8±6.8b 12.8±0.4a
W 6.6±0.5a 22.0±1.0a 2.1±0.1a 28.7±1.8c 5.4±0.8a 54.3±3.3c 13.6±0.6a
N+W 6.4±0.5a 21.3±1.0a 2.2±0.1a 78.3±11.4a 6.6±1.4a 103.8±11.4a 12.7±0.8a
无芒雀麦单播 Smooth bromegrass monoculture
CK 7.2±0.2a 20.6±0.1a 2.1±0.0a 28.6±2.2b 4.7±0.8a 54.7±2.9b 12.5±0.9a
N 6.6±0.2a 21.4±0.5a 2.2±0.0a 81.3±19.4a 6.4±0.9a 121.2±26.1a 13.0±0.9a
W 7.2±0.2a 19.6±1.3a 2.0±0.1a 40.4±9.0b 4.9±0.4a 67.1±10.0b 13.0±1.1a
N+W 7.0±0.3a 20.4±0.8a 2.0±0.1a 55.4±7.6ab 4.8±0.9a 80.9±9.0ab 12.2±0.5a
苜蓿-无芒雀麦混播 Mixed cultivation
CK 6.8±0.2a 21.6±1.2a 2.1±0.1a 29.9±3.0b 5.6±1.0a 55.7±4.7b 12.9±0.5a
N 6.8±0.3a 21.8±1.4a 2.2±0.1a 72.3±7.8a 6.0±1.7a 104.6±6.8a 13.1±0.7a
W 7.0±0.2a 22.5±0.8a 2.2±0.1a 25.8±3.8b 6.0±0.9a 54.4±4.3b 12.4±0.3a
N+W 7.1±0.3a 21.8±2.3a 2.2±0.2a 78.1±10.3a 5.8±0.7a 109.1±11.6a 12.4±0.4a

Fig. 1

Reponses of different SOC fractions to N and water treatments under three forage cultivation types Different lowercase letters indicate significant differences in individual SOC fraction among different N and water addition treatments by Duncan’s multiple range tests (P<0.05)"

Table 3

PLFA concentration (μmol·g-1 soil) in response to N and water treatment under three forage cultivation types"

处理
Treatment		 总磷脂脂肪酸含量
Total PLFA		 革兰氏阳性细菌
Gram-positive bacteria		 革兰氏阴性细菌
Gram-negative bacteria		 真菌
Fungi		 丛枝菌根真菌
AMF		 真菌细菌比
F/B		 革兰氏阳性/
阴性细菌比
G+/G-
苜蓿单播 Alfalfa monoculture
CK 25.1±1.7a 10.2±0.8a 8.3±0.5a 2.5±0.2a 0.86±0.07a 0.137±0.003a 1.23±0.05a
N 21.7±2.7a 8.7±1.1a 7.3±0.8a 2.4±0.4a 0.72±0.09a 0.147±0.006a 1.19±0.03a
W 25.6±3.2a 10.3±1.2a 8.6±1.1a 2.6±0.4a 0.87±0.12a 0.137±0.006a 1.21±0.04a
N+W 26.0±2.3a 10.5±0.9a 8.6±0.8a 2.7±0.3a 0.83±0.09a 0.143±0.002a 1.22±0.03a
无芒雀麦单播 Smooth bromegrass monoculture
CK 23.9±2.6a 9.7±1.1a 7.8±0.8a 2.4±0.3a 0.88±0.10a 0.138±0.008a 1.24±0.02a
N 22.8±2.3a 9.5±0.8a 7.4±0.8a 2.2±0.2a 0.79±0.14a 0.128±0.003a 1.28±0.02a
W 21.7±1.6a 8.8±0.6a 7.1±0.6a 2.3±0.2a 0.75±0.06a 0.143±0.001a 1.25±0.04a
N+W 22.6±1.8a 9.2±0.7a 7.5±0.6a 2.2±0.2a 0.79±0.04a 0.133±0.004a 1.22±0.02a
苜蓿-无芒雀麦混播 Mixed cultivation
CK 22.1±1.1ab 9.1±0.5ab 7.3±0.3ab 2.2±0.1a 0.73±0.03ab 0.132±0.004a 1.25±0.04a
N 25.9±1.6a 10.2±0.7a 8.8±0.5a 2.6±0.2a 0.95±0.07a 0.135±0.003a 1.16±0.02a
W 21.2±2.4ab 8.7±0.9ab 6.9±0.7ab 2.1±0.3a 0.73±0.10ab 0.135±0.005a 1.26±0.05a
N+W 19.8±1.4b 7.8±0.6b 6.8±0.5b 3.0±0.2a 0.69±0.06b 0.144±0.002a 1.15±0.04a

Fig. 2

Responses of extracellular enzyme activity to different N and water treatments under three forage cultivation types Bars indicate mean value ± SE. Different lowercase letters indicate significant differences in individual enzyme activity among different N and water addition treatments by Duncan’s multiple range tests (P<0.05)"

Table 4

Responses of enzyme stoichiometric ratios to N and water treatment under three forage cultivation types"

处理
Treatment		 土壤酶C/N比 Enzyme C/N 土壤酶C/P比 Enzyme C/P
βG/NAG CB/NAG βG/AP CB/AP
苜蓿单播 Alfalfa monoculture
CK 6.5±0.6c 0.33±0.11b 0.58±0.05a 0.033±0.004a
N 12.4±0.6a 0.65±0.11a 0.75±0.06a 0.035±0.006a
W 10.2±1.6ab 0.30±0.02b 0.77±0.14a 0.037±0.006a
N+W 7.7±1.3bc 0.15±0.04b 0.61±0.13a 0.032±0.001a
无芒雀麦单播 Smooth bromegrass monoculture
CK 7.7±0.3a 0.30±0.05a 0.84±0.07a 0.057±0.010a
N 6.4±0.1a 0.32±0.01a 0.70±0.07a 0.052±0.009a
W 8.0±1.0a 0.32±0.07a 1.47±0.33a 0.069±0.006a
N+W 6.8±0.6a 0.28±0.08a 1.25±0.27a 0.087±0.027a
苜蓿-无芒雀麦混播 Mixed cultivation
CK 9.9±0.7a 0.32±0.03ab 0.86±0.12a 0.034±0.003b
N 9.0±0.4a 0.26±0.05b 0.75±0.08a 0.036±0.002b
W 9.5±1.5a 0.40±0.05a 0.91±0.10a 0.038±0.006ab
N+W 8.9±0.5a 0.40±0.04a 1.15±0.20a 0.059±0.012a

Table 5

Pearson correlation coefficient (r) of relative changes (Δ) in PLFA concentration, enzyme activity and SOC fractions under N and water addition"

项目
Item		 全部处理 All treatments 不补水 No water addition 补水 With water addition
Coarse POC Fine POC POC MAOC Coarse POC POC MAOC Coarse POC Fine POC POC MAOC
土壤微生物量 Soil microbial biomass
PLFA总量
Total PLFA		 — 0.424* 0.486* -0.657** — 0.554? -0.679* — 0.679* 0.534? -0.767**
革兰氏阳性细菌
G+		 — 0.426* 0.461* -0.628** — 0.520? -0.616* — 0.682* — -0.781**
革兰氏阴性细菌
G-		 0.390? 0.366? 0.435* -0.629** — 0.538? -0.772** — 0.585? — -0.635*
真菌 Fungi — 0.419* 0.498* -0.623** — 0.574? -0.660* 0.585? 0.564? 0.550? -0.733*
丛枝菌根真菌
AMF		 0.464* 0.378? 0.508* -0.632** — 0.558? -0.710* 0.617* 0.627* 0.563? -0.596?
土壤酶活性 Extracellular enzyme activity
βG — — — — -0.642* — 0.685* — — -0.709* —
CB — — — — -0.651* — 0.711* — — -0.748* —
土壤酶化学计量比 Enzyme stoichiometric ratio
βG/NAG — — — — — — — — — -0.581? —
CB/NAG — — — — -0.691* — 0.607? — — -0.540? —
βG/AP — -0.463* -0.476* 0.404? — — — — -0.797** -0.821** —
CB/AP — — — — — — 0.572? — -0.572? -0.667* —

Fig. 3

Relationships between relative changes (Δ) in POC content and enzyme stoichiometric ratios"

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