淹水条件与植被类型对洱海湖滨带微生物群落特征及脱氮功能基因丰度的影响

廖万雪; 田泽斌; 侯泽英; 祝枫; 储昭升; 袁静

doi:10.12153/j.issn.1674-991X.20240064

淹水条件与植被类型对洱海湖滨带微生物群落特征及脱氮功能基因丰度的影响

doi: 10.12153/j.issn.1674-991X.20240064

湖泊水污染治理与生态修复技术国家工程实验室，中国环境科学研究院

基金项目: 国家重点研发计划重点专项（2021YFC3201003）

详细信息

作者简介:
廖万雪（1999—），女，硕士研究生，主要从事湿地氮转化过程研究，liaowanxue99@163.com

通讯作者:
袁静（1990—），女，助理研究员，博士，主要从事湖滨湿地生态修复及拦截净化技术研究，xyvy-8945@163.com

中图分类号: X14；X524
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出版历程
- 收稿日期: 2024-01-28
- 录用日期: 2024-04-16
- 修回日期: 2024-02-19

Effects of flooding conditions and vegetation types on the microbial community characteristics and the abundance of denitrification functional genes in the ecotone of Lake Erhai

National Engineering Laboratory for Lake Pollution Control and Ecological Restoration, Chinese Research Academy of Environmental Sciences

摘要

摘要:
为探究淹水条件与植被类型对湖滨带微生物群落结构、脱氮功能基因的影响，以洱海湖滨带3种淹水条件（不淹水、间歇淹水、持续淹水）和6种植被类型（草地、林地、林草地、挺水、沉水、无植被）土壤/底泥中的微生物为研究对象，采用16S rRNA基因高通量测序和荧光定量PCR技术，分析淹水条件和植被类型对洱海湖滨带土壤和底泥微生物群落结构以及微生物脱氮功能基因丰度的影响。结果表明：交界区作为湖滨带物质交换与能量流动最频繁的区域，受水位波动影响环境条件复杂，其土壤/底泥中氮循环的重要微生物类群（如绿弯菌门和硝化螺旋菌门等）的相对丰度及α多样性均高于陆向区和水向区；而植被类型主要影响水向区底泥的微生物群落组成和α多样性。微生物脱氮功能基因丰度的差异进一步说明湖滨带脱氮微生物的空间分布特点，陆向区和交界区更高的功能基因丰度说明微生物脱氮活动更强烈。含水率、碳氮比、有机碳和亚硝氮是洱海湖滨带土壤/底泥脱氮过程功能基因丰度的关键影响因素，其浓度变化是淹水条件和植被类型影响脱氮功能基因丰度差异的主要原因。
- 湖滨带 /
- 淹水条件 /
- 植被类型 /
- 微生物 /
- 群落结构
Abstract:
In order to investigate the effects of flooding conditions and vegetation types on the microbial community structure and denitrification function genes of the ecotone zone of Lake Erhai, the microorganisms present in the soil and sediment of the ecotone zone of Lake Erhai were investigated under three flooding scenarios (no-flooding area, intermittent inundation area, constant inundation area) and with six different types of vegetation cover (grassland, woodland, forest-grassland, emergent vegetation, submerged vegetation, and no vegetation). The investigation was conducted using the 16S rRNA gene high-throughput sequencing and fluorescence quantitative PCR technologies to determine changes in microbial community structure and functional gene abundance and the impact of flooding and vegetation cover on soil and sediment microbial communities in the ecotone zone of Lake Erhai. The results showed that as the area with the most frequent material exchange and energy flow in the lakeshore, the interface zone was affected by water level fluctuations, so it became a complex environmental condition. The relative abundance of important microbial groups in nitrogen cycling in this soil/sediment, such as Chloroflexi and Nitrospirota, was higher than in the land and water zone. The vegetation cover mainly affected the microbial community composition and α Diversity in the sediment in the water land. The difference in the abundance of microbial denitrification functional genes revealed the spatial distribution characteristics of denitrification microorganisms in the ecotone zone. A higher abundance of functional genes in landward and interface zones indicated a stronger microbial denitrification activity. The abundance of functional genes during the removal of nitrogen from soil or sediment in the lakeshore of Lake Erhai was affected by several key factors, including water content, carbon-to-nitrogen ratio, organic carbon, and nitrite nitrogen. The difference in the concentration of these factors was the primary cause of the variation in the abundance of functional genes for nitrogen removal under various flooding conditions and vegetation cover.
- ecotone zone /
- flooding condition /
- vegetation type /
- microorganism /
- community structure

HTML全文

图 1 洱海湖滨带采样样地及样点布置示意

Figure 1. Schematic diagram of sampling sites and layout of sample sites along the ecotone zone of Lake Erhai

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图 2 不同区域土壤和底泥的理化性质

Figure 2. Physicochemical properties of soil and sediment in different zones

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图 3 不同区域微生物门水平群落结构

Figure 3. Microbial composition at phylum levels in different zones

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图 4 不同区域微生物群落的Shannon指数、Chao1指数和Simpson指数

Figure 4. Shannon index, Chao1 index and Simpson index of microbial communities in different zones

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图 5 不同区域微生物群落的NDMS分析

Figure 5. NMDS analysis of microbial communities in different zones

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图 6 不同区域微生物脱氮功能基因丰度

Figure 6. Abundance of microbial denitrification functional genes in different zones

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图 7 功能基因丰度与土壤/底泥理化性质的冗余分析

Figure 7. Redundancy analysis of functional gene abundance and physicochemical properties of soil/sediment

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图 8 功能基因丰度与土壤/底泥理化性质的Spearman相关性

注：*表示P<0.05。

Figure 8. Spearman correlation heat map of functional gene abundance with physicochemical properties of soil/sediment

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表 1 定量PCR分析目的基因引物

Table 1. Primers of target genes used in quantitative PCR analysis

目标基因	引物名称	引物序列
AOA amoA	Arch-amoAF	STAATGGTCTGGCTTAGACG
AOA amoA	Arch-amoAR	GCGGCCATCCATCTGTATGT
AOB amoA	amo598f	GAATATGTTCGCCTGATTG
AOB amoA	amo718r	CAAAGTACCACCATACGCAG
narG	narG1960m2f	TA(CT)GT(GC)GGGCAGGA(AG)AAACTG
narG	narG2050m2r	CGTAGAAGAAGCTGGTGCTGTT
napA	napAV17m	TGGACVATGGGYTTYAAYC
napA	napA4r	ACYTCRCGHGCVGTRCCRCA
nirK	nirK876	ATYGGCGGVAYGGCGA
nirK	nirK1040	GCCTCGATCAGRTTRTGGTT
nirS	nirSCd3aF	AACGYSAAGGARACSGG
nirS	nirSR3cd	GASTTCGGRTGSGTCTTSAYGAA
nosZ	nosZ1527F	CGCTGTTCHTCGACAGYCA
nosZ	nosZ1773R	ATRTCGATCARCTGBTCGTT
amx 16S rRNA	amx809F	GCCGTAAACGATGGGCACT
amx 16S rRNA	amx1066R	AACGTCTCACGACACGAGCTG

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表 2 不同区域微生物群落不相似性检验

Table 2. Dissimilarity test of microorganism communities in different zones

对比样地	ADONIS		ANOSIM
对比样地	R²	P	R	P
陆向区×交界区×水向区	0.242	0.001	0.784	0.001
草地×林地（陆向区）	0.153	0.283	0.115	0.241
草地×林草地（交界区）	0.095	0.093	0.228	0.925
挺水×沉水×无植被（水向区）	0.190	0.001	0.447	0.001
注：“×”表示不同淹水条件或植被类型的样地对比。

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