Effects of activated sludge extraction on rhizosphere soil microbial community structure of rice
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摘要:
通过根箱试验，从土壤根际微生物角度揭示活性污泥萃取液（简称萃取液）调控水稻苗期生长和改良土壤理化性质的机理。试验设计不同萃取液配施减氮处理（T1，空白对照；T2，常规施肥对照；T3，减氮30%配施0.2 g/kg萃取液；T4，减氮30%配施0.4 g/kg萃取液；T5，施用8 g/kg萃取液全量替代化肥氮，即减氮75%），研究萃取液配合氮肥减施对水稻根际土壤理化性质和微生物群落结构的影响。结果表明：配施萃取液可显著增加水稻幼苗的叶面积、生物量和叶绿素含量；与常规施肥相比，配施萃取液可以显著提高土壤pH，降低土壤电导率，增加可溶性有机碳含量，降低硝态氮和铵态氮含量。β多样性结果显示，萃取液对土壤微生物性质的影响主要作用在根际及近根际土壤中，不同萃取液处理组与空白及常规施肥对照组间细菌群落结构及多样性存在显著性差异（P<0.05）；萃取液的施用可增加有机污染物降解和碳水化合物代谢相关微生物的相对丰度，如黄色土源菌、Candidatus_Udaeobacter、鞘氨醇单胞菌属，相对丰度分别增加151%~541%、26%~320%和55%~364%。采用FAPROTAX土壤功能预测分析发现，施用萃取液后土壤菌群主要增加的功能为化能异养、有氧化能异养、硝酸盐还原、固氮作用、硝化作用和芳香化合物降解等，这类功能与土壤碳源物质代谢、氮循环及有机污染物降解等行为相关。萃取液施用可通过改善土壤根际及近根际微生物群落结构，增加相关菌群丰度，调控土壤根际及近根际碳、氮循环，进而达到促进水稻幼苗生长和土壤改良的效果。研究结果可为活性污泥的资源化提供新路径。
Abstract:A root box experiment was conducted to reveal the mechanisms by activated sludge extracts regulating rice growth and improving soil physicochemical properties from the perspective of soil rhizosphere microorganisms. The experiment was designed to investigate the effects of the extracts application with nitrogen reduction on rice soil physicochemical properties and rhizosphere microorganisms by conducting different treatments (T1, blank control; T2, conventional fertilizer control; T3, 30% nitrogen reduction with 0.2 g/kg extract solution; T4, 30% nitrogen reduction with 0.4 g/kg extract solution; T5, full replacement of fertilizer nitrogen with 8 g/kg extract solution equivalent to 75% nitrogen reduction). The results showed that the application of extracts significantly increased the leaf area, biomass and chlorophyll content of rice seedlings. Compared with conventional fertilizer applications, the application of extracts could significantly increase soil pH, reduce soil conductivity, increase soil soluble organic carbon and reduce soil nitrate and ammonium nitrogen content. β-diversity results showed that the effect of extracts application on soil microbial properties was more prominent in rhizosphere and near-rhizosphere soils, with significant differences in bacterial community structure and diversity between the different extracts treated groups, blank group and the control group (P<0.05). Application of extracts significantly increased the relative abundance of microorganisms associated with organic pollutant degradation and carbohydrate metabolism, such as Flavisolibacter, Candidatus_Udaeobacter, and Sphingomonas, which increased by 151% to 541%, 26% to 320% and 55% to 364%, respectively. By FAPROTAX soil function prediction analysis, it was found that the main soil microbial communities enhanced after the application of the extracts were chemoheterotrophy, aerobic chemoheterotrophy, nitrate reduction, nitrogen fixation, nitrification and aromatic compound degradation, which were related to the behaviours of soil carbon cycling, nitrogen cycling and organic pollutant degradation. The results of the study indicated that the application of extracts could improve the structure of soil rhizosphere and near-rhizosphere microbial communities, increase the abundance of related bacteria communities, and regulate carbon and nitrogen cycles of rhizosphere and near-rhizosphere soil, thereby promoting the growth of rice seedlings and improving soil characteristics. Therefore, the results of this study can provide a new pathway for the resource utilization of activated sludge.
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图 1 活性污泥萃取液对水稻幼苗生长性状的影响
注：不同字母表示不同处理间存在显著差异（P<0.05）。全文同。
Figure 1. Effect of activated sludge extracts on growth traits of rice seedlings
图 2 活性污泥萃取液对土壤理化性质的影响
Figure 2. Effect of activated sludge extracts on soil physicochemical properties
图 3 各处理不同根际区域细菌群落结构主坐标分析
Figure 3. Principal coordinate analysis of bacterial community structure in different rhizosphere regions for each treatment
图 4 各处理不同根际区域组间细菌群落结构Anosim检验箱线图
Figure 4. Anosim test box plots of bacterial community structure in different rhizosphere regions for each treatment
图 5 各处理间不同根际区域土壤属水平菌群落结构组成分析
注：图中数字为菌群在该组别中所占比例。
Figure 5. Structural composition analysis of genus level bacteria in different rhizosphere regions for each treatment
图 6 土壤理化因子与土壤微生物耦合分析热图
注：*表示显著相关，P<0.05；**表示极显著相关，P<0.01。
Figure 6. Heat map of coupled soil physicochemical factors and soil microbial analysis
图 7 基于FAPROTAX的各处理间不同区域土壤微生物功能预测
Figure 7. Prediction of soil microbial functions in different areas between treatments based on FAPROTAX
表 1 各处理不同根区土壤微生物α多样性指数
Table 1. α diversity index of soil microorganisms in different rhizosphere regions for each treatment
根区 组别 Sobs指数 Chao指数 Ace指数 Simpson指数 Shannon指数 根际 T1 3 701±876.9a 4 178±775.4a 4 453±812.5a 1.0±0.0a 9.5±0.2a T2 4 081±237.6a 4 331±220.7a 4 620±231.2a 1.0±0.0a 8.9±0.4a T3 3 669±245.0a 3 774±269.4a 4 037±281.4a 1.0±0.0a 9.1±0.3a T4 3 832±151.6a 4 109±158.7a 4 377±163.8a 1.0±0.0a 9.4±0.1a T5 3 833±71.1a 4 220±93.0a 4 485±120.8a 1.0±0.0a 9.3±0.1a 近根际 T1 3 572±69.5b 4 040±53.1b 4 311±63.5b 1.0±0.0b 8.7±0.4b T2 3 637±13.0b 4 051±15.3b 4 297±48.9b 1.0±0.0a 9.4±0.3ab T3 3 499±38.7b 3 898±63.2b 4 144±71.6b 1.0±0.0a 9.3±0.1ab T4 3 739±136.7ab 4 246±122.0ab 4 520±138.7ab 1.0±0.0a 9.4±0.0a T5 3 941±124.0a 4 474±132.4a 4 765±124.4a 1.0±0.0a 9.6±0.1a 远根际 T1 3 580±167.8b 3 995±205.5b 4 272±216.0b 1.0±0.0a 8.7±0.4a T2 3 780±95.2ab 4 194±96.6ab 4 473±69.6ab 1.0±0.0a 8.6±0.4a T3 3 657±97.1ab 4 235±90.3ab 4 492±72.9ab 1.0±0.0a 9.3±0.5a T4 3 882±81.9a 4 403±118.5a 4 673±126.2a 1.0±0.0a 9.4±0.1a T5 3 652±33.5ab 4 074±38.6ab 4 292±42.9ab 1.0±0.0a 9.1±0.2a 
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