Effects of Acid Mine Drainage on Physicochemical Factors and Nitrogen-fixing Microorganisms in the Root Zone of Mature Rice

doi:10.13560/j.cnki.biotech.bull.1985.2023-1146

Abstract

Abstract:

【Objective】 Acid mine drainage（AMD）is a kind of wastewater with low pH, high sulfate concentration and heavy metal enrichment. This study is aimed to investigate the abundance and composition of azotobacter community in the root zone of rice at mature stage and its response to AMD, and to elucidate the main driving factors of the change of soil azotobacter community structure. 【Method】 Rice pot experiment was carried out on paddy soil contaminated and uncontaminated by AMD from Tongling mining area of Anhui province, and three different treatments were set up（A: AMD irrigated contaminated soil； B: clean water irrigated contaminated soil; CK: clean water irrigated uncontaminated soil）, nifH gene high-throughput sequencing technology was used to analyze the characteristics of azotobacter community in rice root zone soil under different treatments. 【Result】 The contents of SO₄^2-, NO₃^- and heavy metals in rice root zone soil increased significantly, the soil acidified, and the diversity of azotobacter community decreased. The dominant nitrogen-fixing bacteria in rice root zone soil included Anaeromyxobacter, Geobacter, etc. The number of nitrogen-fixing bacteria enriched in CK treatment was significantly higher than that in A and B treatments, and Verrucomicrobia were mainly enriched in B treatment, while Proteobacteria were mainly enriched in CK treatment. pH、Cu、Pb and Zn were the main factors driving the community structure of azotobacter in rice root zone. Desulfovibrio and Desulfurivibrio with sulfur reduction function have significantly contributed to the change in nitrogen. 【Conclusion】 AMD had significant effects on soil chemistry and azotobacter in the root zone of rice, and restoring clean water irrigation promoted the recovery of azotobacter. The above findings provide new insights and ideas for biological remediation of AMD-contaminated soil.

Key words: AMD, rice root zone soil, root zone nitrogen-fixing microorganism, nitrogen, heavy metal

TIAN Sheng-ni, ZHANG Qin, DONG Yu-fei, DING Zhou, YE Ai-hua, ZHANG Ming-zhu. Effects of Acid Mine Drainage on Physicochemical Factors and Nitrogen-fixing Microorganisms in the Root Zone of Mature Rice[J]. Biotechnology Bulletin, 2024, 40(6): 271-280.

Figures/Tables 8

Table 1 Physical and chemical properties of soil before planting rice

供试土壤 Tested soil	TN/ （g·kg^-1）	NO₃^-/（mg·kg^-1）	NH₄⁺/ （mg·kg^-1）	OM/ （g·kg^-1）	TP/ （g·kg^-1）	pH	SO₄^2-/ （mg·kg^-1）	Cu/ （mg·kg^-1）	Cd/ （mg·kg^-1）	Pb/ （mg·kg^-1）	Zn/ （mg·kg^-1）
污染土AMD contaminated soil	0.97±0.10	6.42±0.60	7.52±0.90	13.01±1.72	0.72±0.07	4.68±0.05	282.41±6.29	151.82±5.94	2.49±0.11	159.02±5.19	210.30±5.39
未污染土Uncontaminated soil	0.98±0.90	3.89±0.51	8.05±0.64	8.29±0.44	0.67±0.01	6.18±0.07	132.87±4.79	20.85±0.57	1.99±0.13	28.30±0.94	90.96±2.26

Fig. 1 Schematic diagram of the test

Fig. 2 Characteristics of soil physical and chemical factors under different treatments A: AMD irrigated contaminated soil; B: clean water irrigated contaminated soil; CK: clean water irrigated uncontaminated soil. Different lowercase letters indicate significant difference between different treatments (P < 0.05). The same below

Fig. 3 Differences in diversity and community structure of nitrogen-fixing microorganisms in rice root zone soil under different treatments a: Box plots of Chao1 and Shannon index diversity of azotobacter in rice root zone soil samples under different treatments; b: PcoA analysis of nitrogen-fixing microbial communities in rice root zone

Fig. 4 Differences in community composition of nitrogen-fixing microorganisms in rice root zone soil under different treatments a: Distribution of azotobacter in rice root zone soil samples under different treatments; b: LEfSe was based on community difference analysis among different treatment groups A, B and CK, and only groups with LDA values > 2 were clearly displayed

Fig. 5 Effects of environmental factors on nitrogen-fixing microorganisms at OTUs level

Fig. 6 Correlation between relative abundances of azotobacter genera and environmental factors *: P< 0.05; **: P < 0.01

Table 1 Stepwise regression analysis of nutrient factors in rice root zone soil and each azotobacter genus

养分因子Nutrient factor	回归方程Regression equation	R²	P
TN	Y=1.778-4225.623×Sinorhizobium-47.255×Desulfovibrio+456.497×Azoarcus-97.577×Azospira+1.902×Geobacter	1	0.000***
NO₃^-	Y=8.099-1661.173×Desulfurivibrio	1	0.003***
OM	Y=14.024-2588.656×Rhodopseudomonas	0.557	0.013**
TP	Y=89.211-97715.976×Xanthobacter-25902.225×Pelomonas-693.042×Desulfovibrio+1097.825×Rubrivivax+13487.397×Desulfobulbus	0.994	0.000***
SO₄^2-	Y=-357.746+614469.605×Frankia+91743.681×Sideroxydans	0.903	0.000***

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