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【目的】分析长期有机无机肥配施对土壤真菌丰度、多样性及其群落特征的影响,探讨东北黑土真菌群落变化与施肥的相关性,为进一步调节土壤微生物结构,改善其生态功能提供参考依据。【方法】依托黑龙江省农业科学院36年长期定位试验站,选取4个不同施肥处理:不施肥处理(CK)、有机肥处理(M)、氮磷钾无机肥处理(NPK)和有机肥配施无机肥处理(MNPK)的耕作层土壤为研究对象,以真菌ITS基因为分子标靶借助q PCR技术和Illumina Miseq高通量测序平台,研究不同施肥处理对黑土中真菌群落丰度、多样性和组成的影响,并与土壤化学性质进行偶联分析,揭示群落与施肥的相关性。【结果】长期施用无机肥显著降低土壤p H,而有机无机配施可以有效缓解土壤酸化。NPK处理的ITS基因丰度显著高于MNPK;MNPK处理的细菌/真菌比值(26.91×104)显著大于NPK,各处理比值由高到低为MNPK>M>CK>NPK。细菌/真菌比值与土壤p H正相关;MNPK处理的真菌α多样性指数值略大于NPK。Ascomycota和Zygomycota为土壤中主要真菌门,不同施肥处理之间真菌组成的相对丰度存在显著差异,对照处理Ascomycota的相对丰度为45.35%,MNPK和NPK处理分别为50.93%和56.16%。有机肥有利于降低病原真菌相对丰度,具有高度侵染性的Cochliobolus在MNPK(0.41%)和M(0.39%)中的相对丰度显著小于CK(3.25%)和NPK(2.08%)。CCA分析表明,土壤理化性质共解释土壤真菌群落结构变化的73.3%,有效磷(贡献量为32.4%,P=0.002)、铵态氮(贡献量为14.8%,P=0.01)和硝态氮(贡献量为16.2%,P=0.048)是3个重要的影响因子。【结论】不同施肥条件下土壤真菌丰度、多样性,以及菌群组成特征不同。与无机肥相比,有机肥无机肥配施能够有效改善真菌群落结构,降低真菌的丰度,增加真菌多样性,并提高土壤p H,减缓土壤酸化。土壤有效磷、铵态氮和硝态氮含量是影响黑土土壤真菌群落结构变化重要因素。
【Objective】 The objective of this study was to analyze the effects of long-term organic and inorganic fertilizers on the abundance, diversity and community characteristics of soil fungi, and to explore the correlation between changes of fungal community and fertilization in the black soils of northeastern China so as to provide reference for further regulating soil microbial structure and improving its ecological functions in accordance with. 【Method】 Based on 36-year long-term experiment station of Heilongjiang Academy of Agricultural Sciences, four different fertilization treatments were selected: CK, M, NPK and organic manure Inorganic fertilization (MNPK) tillage soil was used as the research object. The fungal ITS gene was used as a molecular target to study the effects of different fertilization treatments on the abundance and diversity of fungal communities in black soil by q PCR and Illumina Miseq high-throughput sequencing platform Composition of the soil chemical properties and coupling analysis to reveal the community and fertilization relevance. 【Result】 Long-term application of inorganic fertilizers significantly reduced soil p H, while organic and inorganic fertilizers were effective in alleviating soil acidification. The abundance of ITS gene of NPK was significantly higher than that of MNPK. The ratio of MNPK to bacteria / fungus (26.91 × 104) was significantly higher than that of NPK, and the ratios of MNPK> M> CK> NPK were the highest to the lowest. The ratio of bacteria to fungi was positively correlated with p H of soil. The value of α diversity index of fungi treated with MNPK was slightly larger than that of NPK. Ascomycota and Zygomycota were the major fungi in soil. The relative abundance of fungi in different fertilization treatments was significantly different. The relative abundance of Ascomycota was 45.35%, and MNPK and NPK were 50.93% and 56.16%, respectively. Organic fertilizer was beneficial to reduce the relative abundance of pathogenic fungi, and the highly invasive Cochliobolus was significantly less abundant in MNPK (0.41%) and M (0.39%) than CK (3.25%) and NPK (2.08%). CCA analysis showed that soil physical and chemical properties explained a total of 73.3%, available phosphorus (32.4%, P = 0.002), ammonium nitrogen (14.8%, P = 0.01) and nitrate nitrogen (Contribution of 16.2%, P = 0.048) are three important influencing factors. 【Conclusion】 The soil fungi abundance, diversity and the characteristics of the flora composition are different under different fertilization conditions. Compared with inorganic fertilizers, organic manure combined with inorganic fertilizers can effectively improve the fungal community structure, reduce fungal abundance, increase fungal diversity, and improve soil p H, slowing soil acidification. Soil available phosphorus, ammonium nitrogen and nitrate nitrogen content is an important factor affecting the community structure of black fungi in the soil.