土壤改良剂对再生水滴灌根际土壤菌群多样性及病原菌和抗生素抗性基因丰度的影响

Effects of Soil Amendments on the Bacterial Diversity and Abundances of Pathogens and Antibiotic Resistance Genes in Rhizosphere Soil Under Drip Irrigation with Reclaimed Water

基于再生水农业灌溉利用引发的人体健康和环境风险,通过施用土壤改良剂揭示再生水灌溉根际土壤菌群组成与多样性变化特征,并探讨土壤改良剂对根际土壤病原菌和抗生素抗性基因丰度变化的影响规律,对于土壤改良剂的合理施用具有指导意义.采用高通量测序技术和定量PCR检测方法,研究了生物质炭、生物有机肥、腐植酸、松土精和玉米酒糟对再生水滴灌根际土壤细菌群落多样性及特定基因丰度的影响.结果表明,生物质炭处理显著增加根际土壤有机质和总氮含量;生物有机肥处理显著增加EC值和有机质含量;玉米酒糟处理显著增加EC值、总氮和总磷含量(P<0.05).除生物质炭处理外,其他各处理均能显著降低根际土壤pH(P<0.05). 5种改良剂处理下根际土壤细菌群落组成与多样性在纲和属水平上较相似,但其相对丰度存在差异.α-Proteobacteria、γ-Proteobacteria、 Bacteroidia、 Actinobacteria、 Acidimicrobiia和Anaerolineae为所有处理中的优势菌纲,优势菌属组成包括:Pseudomonas、Sphingobium、Sphingomonas、Cellvibrio、Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium、Flavobacterium和Algoriphagus(相对丰度>1%).环境因子关联分析表明,根际土壤细菌群落组成与pH、 EC、总氮和总磷含量之间存在较强的关联.病原菌与抗生素抗性基因的检出丰度分别在10~3~10~7 copies·g和10~4~10~8 copies·g.改良剂对病原菌和抗生素抗性基因检出水平存在较大差异,生物有机肥、松土精和玉米酒糟处理均会导致部分抗生素抗性基因丰度显著增加,而腐植酸和玉米酒糟处理下丁香假单胞菌、茄科雷尔氏菌和大肠菌群丰度显著降低(P<0.05).弓形菌、蜡样芽孢杆菌、成团泛菌和粪拟杆菌与四环素类(tetA、tetB、tetO和tetQ)、磺胺类(sul1)和红霉素类(ermB和ermC)抗性基因丰度存在显著相关性.研究认为监测再生水灌溉下农业环境中病原菌和抗生素抗性基因的同时,也要关注土壤改良剂的合理施用避免加剧生物污染的散播.

Due to reclaimed water, irrigation can cause human health and environmental risks. Soil amendments are applied to reveal the abundance of pathogens and antibiotic resistance genes in rhizosphere soil irrigated by reclaimed water and to better understand the effects of environmental factors on the rhizosphere soil bacterial composition, which has guiding significance for the reasonable use of soil amendments. In this study, the effects of biochar, bioorganic fertilizer, humic acid, loosening soil essence, and corn vinasse on bacterial community diversity and certain gene abundances in rhizosphere soil under drip irrigation with reclaimed water were studied using high-throughput assays and quantitative PCR. The results showed that biochar significantly increased pH, organic matter, and total nitrogen contents in the rhizosphere soil. The corn vinasse significantly decreased soil pH and increased the contents of total nitrogen and total phosphorus but significantly increased the soil EC value(P<0.05). The effects of the five soil amendments on the α-diversity of rhizosphere bacteria were not significantly different. The bacterial community structure and diversity of rhizosphere bacteria were similar at different taxonomic levels, but their relative abundance was different. α-Proteobacteria, γ-Proteobacteria, Bacteroidia, Actinobacteria, Acidimicrobiia, and Anaerolineae were the dominant bacteria in all treatments. The dominant genera consisted of Pseudomonas, Sphingobium, Sphingomonas, Cellvibrio, Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium, Flavobacterium, and Algoriphagus(relative abundance>1%). Correlation analysis of environmental factors showed that the composition of the rhizosphere bacterial community was strongly correlated with pH, EC, total nitrogen, and total phosphorus content. The abundances of pathogenic bacteria and antibiotic resistance genes were 10~3-10~7 copies·gand 10~4-10~8 copies·g, respectively. There were significant differences in the detection levels of pathogens and antibiotic resistance genes. Bioorganic fertilizer, loosening soil essence, and corn vinasse significantly increased the abundances of some antibiotic resistance genes, whereas humic acid and corn vinasse significantly decreased the abundances of Pseudomonas syringae, Ralstonia solanacearum, and total coliforms(P<0.05). A significant correlation was found between pathogens(Arcobacter, Bacillus cereus, Pantoea agglomerans, and Fecal bacteroidetes) and antibiotic resistance genes(tetA, tetB, tetO, tetQ, sul1, ermB, and ermC). In conclusion, while monitoring pathogens and antibiotic resistance genes in the agricultural environment under reclaimed water irrigation, attention should be paid to the rational application of soil amendments to avoid exacerbating the spread of biological contamination.