基于分子生态学网络分析松嫩退化草地土壤微生物群落对施氮的响应

doi:10.3864/j.issn.0578-1752.2020.13.012

Abstract

Abstract:

【Objective】 Nitrogen input affects the sustainability of the global grassland ecosystem. Paying attention to the soil microbial community and its molecular ecological network can provide a theoretical basis for grassland degradation restoration. 【Method】 Taking Songnen degraded Leymus chinensis grassland as the research object, the molecular ecological network of soil microbial community was constructed by applying high-throughput sequencing and random matrix network construction with and without nitrogen treatment. To explore the effects of nitrogen management on the soil microbial community structure and network in degraded Leymus chinensis grassland, the key microbial changes in the microbial network structure under the condition of nitrogen addition was studied, and the interaction between microorganisms during the process were investigated, and the conditions for external nitrogen addition key points and regularity of soil bacterial dynamic change were analyzed. 【Result】 At the level of phylum classification, there were 22 bacterial phylum in nitrogen-applied grassland and 23 without nitrogen. The 7 phylum were the dominant phylum of the nitrogen-applied and non-nitrogen-applied grasslands. Among them, Proteobacteria was the phyla containing the largest number of OTUs, accounting for about 30.46% of the total sequence. The next largest genus was about 30.15% of the total sequence. The genus Gemmatimonadetes was the third genus containing OTUs, accounting for 8.14% of the total sequence. Actinomycete accounted for about the total 6.15% of the sequence, while Chloroflexi, Bacteroidetes and Nitrospirae accounted for 17.16% of the total sequence. The relative abundances of Proteobacteria, Actinomycota and Bacteroides in soil microorganisms in nitrogen-applied grassland were significantly higher than those in non-nitrogen applied grassland soil; The relative abundances of Proteobacteria, Actinomycete and Bacteroidetes were significantly higher than the soils of nitrogen-applied grassland (P<0.01), and no significant difference was found between the nitrogen application and non-nitrogen treatment of other bacteria. The forward connection ratio, the average path length, the average clustering coefficient, and the modularity of the characterizing network were all significantly lower than the nitrogen-free treatment (P<0.001). In the molecular ecological network of soil, there were 16 modular hubs without nitrogen treatment (Zi>2.5, Pi≤0.62), and there were 6 modular hubs under nitrogen treatment, all of which belong to Acidobacteria, Gemmatimonadetes and Actinomycete. Nitrogen application led to changes in soil microbial species relationships, which in turn changed the overall soil ecological network. 【Conclusion】 Nitrogen application reduced the complexity and tightness of soil network structure of degraded grassland, and reduced the relative abundance of Acidobacteria and Chloroflexi in degraded grassland, while which improved the relative abundance of Proteobacteria, Actinomycete and Gemmatimonadetes. The number of microbial key species (OTU) in soil decreased from 16 (no nitrogen application) to 6 and there was no overlapping OTU in both soils, indicating that nitrogen application regulated key species of its community network and thus changes its molecular ecological network.

Key words: nitrogen application, degraded grassland, soil microbe, molecular ecology network

ZHU RuiFen,LIU JieLin,WANG JianLi,HAN WeiBo,SHEN ZhongBao,XIN XiaoPing. Molecular Ecological Network Analyses Revealing the Effects of Nitrogen Application on Soil Microbial Community in the Degraded Grasslands[J].Scientia Agricultura Sinica, 2020, 53(13): 2637-2646.

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References 30

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处理 Treatment	pH	土壤全氮 STN (g·kg^-1)	土壤全磷 STP (g·kg^-1)	土壤铵态氮 SNH₄ (mg·kg^-1)	土壤硝态氮 SNO₃ (mg·kg^-1)	土壤速效磷 SAP (mg·kg^-1)
F	7.95a	1.71a	0.53b	17.80a	21.62a	10.45a
NF	8.09a	1.31b	0.60b	12.61b	14.75b	10.49a

样品名称Sample ID	双端序列 PE reads	原始序列 Raw tags	优化序列 Clean tags	有效序列 Effective tags	平均序列长度 Average length (bp)	GC (%)	Q20 (%)	Q30 (%)	Effective (%)
NF	47023	44654	39465	38754	421	57.14	97.15	94.39	82.41
F	66659	63320	56208	55106	420	56.61	97.26	94.61	82.67

样品名称 Sample ID	变形菌门 Proteobacteria	酸杆菌门 Acidobacteria	芽单胞菌门Gemmatimonadetes	放线菌 Actinomycete	绿弯菌门Chloroflexi	拟杆菌门Bacteroidetes	硝化螺旋菌门Nitrospirae
NF	282b	280a	76a	57b	37b	26b	14b
F	316a	258b	52b	89a	31a	30a	22a

处理Treatment	阈值 Threshold	节点数 Total node	连接数 Total links	正向连接比 Positive link percentage (%)	平均连通度 Connectivity	平均路径长度 Average path distance	平均聚集系数 Clustering coefficient	模块性 Modularity
F	0.9	1872	19873	53.2*	23.6*	2.33*	0.435*	0.46*
NF	0.9	1913	20389	69.4*	28.7*	2.89*	0.462*	0.41*

No.	OTUs	关键OTUs Key OTUs	样品名称Sample ID
1	OTU76	c_Subgroup_6;Unclassified; Unclassified; Unclassified	NF
2	OTU545	c_Chloroflexia;o_Chloroflexales; f_Roseiflexaceae; g_Roseiflexus	NF
3	OTU1682	g_Chthoniobacter; s_uncultured_Verrucomicrobia_bacterium	NF
4	OTU1975	g_Bryobacter; s__uncultured_bacterium_g_Bryobacter	NF
5	OTU3589	g_Roseomonas	NF
6	OTU1174	g_Haliangium	NF
7	OTU300	g_uncultured_bacterium_f_0319-6A21; s_uncultured_ bacterium_f_0319-6A21）	NF
8	OTU2179	g_uncultured_bacterium_f_Acidobacteriaceae_ [Subgroup_1]	NF
9	OTU688	g_uncultured_bacterium_c_Gitt-GS-136; s_uncultured_bacterium_c_Gitt-GS-136	NF
10	OTU5024	c_Subgroup_6; Unclassified; Unclassified; Unclassified; Unclassified	NF
11	OTU1827	f_Geobacteraceae; Unclassified; Unclassified	NF
12	OTU6317	g__uncultured_bacterium_f_Caldilineaceae; s__uncultured_bacterium_f_Caldilineaceae	NF
13	OTU828	c_Subgroup_6	NF
14	OTU1069	g_uncultured_bacterium_f_Acidimicrobiaceae	NF
15	OTU3349	c_OM190; Unclassified; Unclassified; Unclassified; Unclassified	NF
16	OTU1830	s_uncultured_bacterium_c_BD2-11_terrestrial_group	NF
17	OTU1179	s_uncultured_bacterium_f_Chitinophagaceae	F
18	OTU1954	c_Betaproteobacteria; o_SC-I-84; Unclassified; Unclassified; Unclassified	F
19	OTU2194	g_Gemmatirosa; s_uncultured_bacterium_g_Gemmatirosa	F
20	OTU532	g_uncultured_bacterium_f_MWH-CFBk5;s_uncultured_bacterium_f_MWH- CFBk5	F
21	OTU3703	c_MB-A2-108; Unclassified; Unclassified; Unclassified; Unclassified	F
22	OTU630	c_Nitrospira; o_Nitrospirales; f_Nitrospiraceae; g_Nitrospira	F

Molecular Ecological Network Analyses Revealing the Effects of Nitrogen Application on Soil Microbial Community in the Degraded Grasslands

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