摘要:

目的

固氮微生物是土壤中重要的功能微生物，其多樣性和群落組成變化能夠影響土壤氮素固定與氮循環(huán)過程，探究不同秸稈還田方式對根際土壤固氮菌多樣性和群落組成的影響機制具有重要意義。

方法

基于中國科學(xué)院鷹潭紅壤生態(tài)實驗站花生單作系統(tǒng)不同秸稈還田長期定位試驗，設(shè)置不施肥對照(CK)、單施化肥(NPK)、NPK肥+秸稈還田(NPKS)、NPK肥+秸稈豬糞配施(NPKSM)和NPK肥+秸稈生物炭(NPKB) 5個處理，利用高通量測序技術(shù)，分析不同秸稈還田方式下根際固氮菌多樣性和群落組成的變化特征。

結(jié)果

秸稈還田處理下土壤有機碳(SOC)、速效鉀、全磷、有效磷、全氮含量提升，其中以NPK肥+秸稈豬糞配施(NPKSM)處理效果最佳。秸稈還田增加了固氮微生物多樣性，并顯著改變其群落組成，在綱水平上固氮菌以α-變形菌綱(Alphaproteobacteria，82.5%)為優(yōu)勢類群；在屬水平上以慢生根瘤菌屬(Bradyrhizobium，51.9%)為優(yōu)勢類群。土壤有效磷是影響固氮菌多樣性指數(shù)的主要因素，而土壤pH、SOC、速效鉀、全磷、有效磷、全氮和銨態(tài)氮是影響固氮菌群落組成的主要因素。結(jié)構(gòu)等式方程研究結(jié)果表明土壤有效磷和全氮通過改變δ-變形菌綱(Deltaproteobacteria)的相對豐度和固氮菌群落組成間接影響花生產(chǎn)量。

結(jié)論

秸稈還田顯著提升了土壤肥力，土壤有效磷是根際固氮菌多樣性和群落組成改變、花生產(chǎn)量提高的重要驅(qū)動因素，通過提高Deltaproteobacteria的相對豐度促進(jìn)了花生增產(chǎn)。本研究為建立合理的秸稈還田措施，增強生物固氮潛力以及提升紅壤肥力與健康提供科學(xué)依據(jù)。

Abstract:

Objectives

Nitrogen-fixing bacteria are an important functional group in the soil microbial community. The changes in the diversity and structure of nitrogen-fixing bacterial community can affect soil nitrogen fixation and nitrogen cycle dynamics. Here, we explored the mechanism of the diversity and structure of rhizosphere nitrogen-fixing bacterial community in regulating peanut yield under the different straw returning treatments.

Methods

We conducted the long-term field experiment with different types of straw returning in peanut mono-cropping system at the Yingtan National Agroecosystem Field Experiment Station of the Chinese Academy of Sciences in Jiangxi. The field experiment included five treatments: no fertilization (CK), conventional NPK (NPK), NPK with straw (NPKS), NPK with straw and pig manure (NPKSM), and NPK with straw biochar (NPKB). The illumina sequencing of nifH gene was used to investigate the diversity and structure of nitrogen-fixing bacterial community in the rhizosphere.

Results

The treatments with straw returning significantly promoted soil fertility including soil organic carbon (SOC), available K, total P, available P, total N, with the highest values under the NPKSM treatment. Compared with the CK and NPK treatments, the NPKS and NPKSM treatments significantly increased the diversity of soil nitrogen-fixing bacteria and shaped community composition. Alphaproteobacteria (82.5%) was the dominant class in the nitrogen-fixing bacteria community, and Bradyrhizobium (51.9%) was the dominant genus. Soil available P was the most important predictor of the nitrogen-fixing bacterial diversity, while pH, SOC, available K, total P, available P, total N and ammonia nitrogen (NH4+-N) were the significant predictors of the nitrogen-fixing bacterial community composition. Structural equation modeling suggested that available P and total N was indirectly associated with peanut yield by changing the structure of nitrogen-fixing bacterial community.

Conclusions

Straw returning significantly improved soil fertility, and available P was an important factor in driving the nitrogen-fixing bacterial community composition and peanut yield by enhancing the relative abundance of Deltaproteobacteria. Taken together, our study provides the scientific basis for establishing reasonable measures of straw returning to promote the potential of biological nitrogen fixation, and to improve the fertility and health of red soil.

圖  1   不同秸稈還田方式對花生產(chǎn)量的影響

[注（Note）：CK—不施肥對照 No fertilization; NPK—單施常規(guī)化肥 Only conventional NPK; NPKS—NPK肥+秸稈還田 NPK with straw; NPKSM—NPK肥+秸稈豬糞配施 NPK with straw and pig manure; NPKB—NPK肥+秸稈生物炭 NPK with straw biochar. 柱上不同字母表示處理間差異達(dá)到顯著水平 (P < 0.05) Different letters above the bars indicate significant difference among treatments (P < 0.05). ]

Figure  1.   Effects of different straw returning methods on peanut yield

圖  2   土壤固氮菌多樣性的Shannon指數(shù)(a)與Chao1指數(shù)(c)和對Shannon指數(shù)(b)和Chao1指數(shù)(d)的隨機森林分析

[注（Note）：CK—不施肥對照 No fertilization; NPK—單施常規(guī)化肥 Only conventional NPK; NPKS—NPK肥+秸稈還田 NPK with straw; NPKSM—NPK肥+秸稈豬糞配施 NPK with straw and pig manure; NPKB—NPK肥+秸稈生物炭 NPK with straw biochar. AK—速效鉀Available K; TP—全磷 Total P; AP—有效磷 Available P; TN—全氮 Total N; SOC—土壤有機碳 Soil organic C. 柱上不同字母表示處理間差異達(dá)到顯著水平 (P < 0.05) Different letters above the bars indicate significant difference among treatments (P < 0.05). *—P < 0.05.]

Figure  2.   Shannon (a) and Chao1 indices (c) of soil nitrogen-fixing bacterial community, and random forest modelling analysis of soil properties on Shannon (b) and Chao1 indices (d)

圖  3   土壤固氮菌群落的主坐標(biāo)分析(PCoA) (a)和隨機森林分析(b)

[注（Note）：CK—不施肥對照 No fertilization; NPK—單施常規(guī)化肥 Only conventional NPK; NPKS—NPK肥+秸稈還田 NPK with straw; NPKSM—NPK肥+秸稈豬糞配施 NPK with straw and pig manure; NPKB—NPK肥+秸稈生物炭 NPK with straw biochar. AK—速效鉀 Available K; TP—全氮 Total P; AP—有效磷 Available P; TN—全氮Total N; SOC—土壤有機碳 Soil organic carbon. **—P < 0.01, *—P < 0.05.]

Figure  3.   Primary co-ordinate analysis (PCoA) of soil nitrogen-fixing microbial communities (a) random forest modelling analysis of community composition (b)

圖  4   土壤固氮菌在門水平 (a)和屬水平(b)的組成及相對豐度

[注（Note）：CK—不施肥對照 No fertilization; NPK—單施常規(guī)化肥 Only conventional NPK; NPKS—NPK肥+秸稈還田 NPK with straw; NPKSM—NPK肥+秸稈豬糞配施 NPK with straw and pig manure; NPKB—NPK肥+秸稈生物炭 NPK with straw biochar. AK—速效鉀 Available K; TP—全氮 Total P; AP—有效磷 Available P; TN—全氮Total N; SOC—土壤有機碳 Soil organic carbon. **—P < 0.01, *—P < 0.05.]

Figure  4.   Relative abundance of soil nitrogen-fixing bacterial community at the phylum level (a) and genus level (b)

圖  5   土壤化學(xué)性質(zhì)與固氮菌多樣性指數(shù)、群落組成及優(yōu)勢類群相對豐度之間相關(guān)性

[注（Note）：AK—速效鉀 Available K; TP—全磷 Total P; AP—有效磷 Available P; TN—全氮 Total N;SOC—土壤有機碳 Soil organic carbon. *—P < 0.05; **—P < 0.01.]

Figure  5.   Correlations among soil chemical properties, the diversity and composition of nitrogen-fixing bacterial community, and relative abundance of dominant groups

圖  6   生物因素和非生物因素對花生產(chǎn)量影響的隨機森林模型和結(jié)構(gòu)等式模型分析

[注（Note）：Comp.—群落組成 Community composition; AP—有效磷 Available P; TP—全磷 Total P; TN—全氮 Total N; AK—速效鉀 Available K; SOC—土壤有機碳 Soil organic carbon. *—P < 0.05; **—P < 0.01; ***—P < 0.001. 箭頭旁邊的數(shù)字代表路徑系數(shù)；線條粗細(xì)表示相關(guān)性大小 The number next to the arrow represents the path coefficient; the thickness of the line indicates the magnitude of the correlation.]

Figure  6.   Effects of biotic and abiotic factors on peanut yield based on random forest modelling and structural equation modelling

表  1   不同秸稈還田方式對土壤化學(xué)性質(zhì)的影響

Table  1   Effects of different straw returning methods on soil chemical properties

處理
TreatmentpH有機碳
Organic C
(g/kg)全氮
Total N
(g/kg)全磷
Total P
(g/kg)有效磷
Available P
(mg/kg)速效鉀
Available K
(mg/kg)銨態(tài)氮
NH4+-N
(mg/kg)硝態(tài)氮
NO3?-N
(mg/kg) CK4.61±0.02 a3.41±0.50 d0.47±0.07 d0.26±0.01 c0.66±0.60 b74.17±5.20 c15.74±1.86 a2.05±0.68 aNPK4.48±0.03 b4.39±1.05 cd0.57±0.06 cd0.46±0.05 b7.27±1.68 b123.33±10.10 bc18.07±5.25 a4.18±2.03 aNPKS4.56±0.03 ab6.42±0.88 bc0.70±0.04 bc0.49±0.05 b11.08±1.73 b179.17±12.83 ab16.83±0.54 a4.70±1.90 aNPKSM4.62±0.07 a8.10±1.38 ab0.92±0.04 a0.76±0.12 a61.24±15.30 a218.33±43.11 a17.49±1.37 a6.33±3.53 aNPKB4.59±0.05 ab9.46±0.68 a0.78±0.08 ab0.48±0.05 b8.89±1.93 b171.67±13.77 ab14.56±0.81 a5.39±1.74 a 注（Note）：CK—不施肥對照 No fertilization; NPK—單施常規(guī)化肥 Only conventional NPK; NPKS—NPK肥+秸稈還田 NPK with straw; NPKSM—NPK肥+秸稈豬糞配施 NPK with straw and pig manure; NPKB—NPK肥+秸稈生物炭 NPK with straw biochar. 表中數(shù)據(jù)均為平均值 ± 標(biāo)準(zhǔn)差。同列數(shù)據(jù)后不同字母表示不同處理間差異顯著(P < 0.05) Data in table are means±standard deviation (n = 3). Values followed by different letters indicate significant difference among treatments (P < 0.05). [1]

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