黑土细菌及真菌群落对长期施肥响应的差异及其驱动因素

doi:10.3864/j.issn.0578-1752.2018.05.010

摘要/Abstract

摘要： 【目的】研究长期施肥对黑土细菌和真菌群落结构影响差异，探索黑土肥力对长期施用化肥和有机肥响应差异的生物学机制，为黑土的肥力培育和合理施肥提供科学理论依据。【方法】基于35年的长期定位施肥试验，采用定量PCR方法和Miseq高通量测序技术，分析长期不施肥（CK）、氮肥（N）、有机肥（M）和有机无机配施肥（MN）处理下，黑土细菌及真菌的数量、群落结构和多样性的差异。同时结合土壤理化性状，探究不同施肥条件下细菌和真菌群落变化的环境驱动因子。【结果】N处理对土壤细菌的数量没有显著影响，但使其群落多样性降低了13.2%—48.5%。N处理使真菌的数量增加了24倍，多样性降低了4.6%—80.3%。与N处理相比，MN处理使细菌数量和多样性分别增加了2倍和7.7%—46.6%，而真菌的数量虽降低了14.2%，但其多样性提高了62%—237%。单施氮肥增加了土壤细菌酸杆菌门（Acidobacteria）中的Acidobacteria_Gp1、Gp3及变形菌门（Proteobacteria）中的α-Proteobacteria的相对丰度，并使土壤真菌中伞菌纲（Agaricomycetes）的相对丰度增加了41倍。与N处理相比，MN处理下细菌的各主要类群丰度未发生显著变化，但M处理下土壤细菌中的α-Proteobacteria、Acidobacteria_Gp1和Gp3丰度分别显著降低了26、97和81个百分点，Acidobacteria_Gp4、Gp6和Plancomycetes的丰度分别显著增加了11倍、9倍和2倍。细菌群落结构在MN与N处理之间无显著差异，明显区别于CK和M处理，pH为主要驱动因素，其阈值为6.07；真菌群落结构在CK、M和MN处理下相似，显著区别于N处理，两组处理之间差异由速效钾含量（125.5 mg·kg^-1）驱动。另外，有机质含量对于细菌和真菌群落均是重要的驱动因素，但调控细菌群落结构的阈值为28.4 g·kg^-1，而驱动真菌群落结构的阈值为30.8 g·kg^-1。【结论】黑土细菌对有机肥的响应较强，而真菌对化肥更为敏感。长期施用化肥会刺激土壤中嗜酸细菌和真菌的生长，而有机无机肥配施可提高土壤微生物群落多样性，刺激有益菌的生长。土壤pH和有效钾含量分别是调控细菌和真菌群落结构的重要影响因素，在黑土肥力培育中应引起充分的重视。

关键词: 细菌, 真菌, 长期施肥, mesiq测序, qPCR, 黑土

Abstract: 【Objective】This study was conducted to explore the bacterial and fungal community responses to long-term chemical fertilizer and manure application, and thus to clarify the biological mechanism of fertilization on soil fertility in black soils. It aims to provide a theoretical support for rational application of fertilizer and black soil fertility improvement. 【Method】The study was conducted on the long-term fertilization (35 years) experiment at Harbin, China. Soil samples were collected from the following four treatments: control, non-fertilization (CK), chemical nitrogen fertilizer (N), manure only (M) and M plus N (MN). Miseq pro-sequencing and qPCR technology were used to find out the difference between bacterial and fungal communities. In combination with soil properties, we analyzed the driving factors for bacterial and fungal community by multivariate statistical analysis. 【Result】Compared with CK, N treatment significantly decreased bacterial diversity and fungal diversity by 13.2%-48.5% and 4.6%-80.3%, respectively, while fungal abundance was increased by 24 times. Applied manure to N fertilization enhanced bacterial abundance and bacterial diversity by 2 times and 7.7%-46.6%, respectively. However, fungal quantity was declined by 14.2% and fungal diversity was increased by 62%-237%, comparing MN with N only fertilizer. In comparison with CK, the abundance of Acidobacteria_Gp1, Gp3 and α-Proteobacteria (bacterial classes) were significantly increased, and Agaricomycetes (fungal class) even was enhanced by 41 times with the addition of N. Compared with N treatment, the bacterial abundance kept constant for MN treatment, while the abundance of α-Proteobacteria, Acidobacteria_Gp1 and Gp3 were decreased, and Acidobacteria_Gp4, GP6 and Plancomycetes were increased for M treatment. Bacterial community structure for MN and N treatments appeared similar, which were significantly different from CK and M treatments. Fungal community structure for CK, M and MN treatments were similar and significantly different from N treatment. Soil pH (6.07) and available potassium (125.5 g·kg^-1) were the principal factors for the difference of bacterial community and fungal community, respectively. Soil organic matter explained both bacterial and fungal community structure alternations while the criteria was different as 28.4 g·kg^-1for bacteria and 30.8 g·kg^-1 for fungi. 【Conclusion】Therefore, our results indicate that bacteria was sensitive to the manure, and fungi was sensitive to N fertilizer application. Long term N application stimulated the growth of acidophilic microbe, while addition of manure to N enhanced microbial diversity and promoted the growth of beneficial microorganism. Soil pH and available potassium were the principal factors driving bacterial and fungal community structure, respectively. Further detailed study is required on this aspect for the improvement of black soil quality.

Key words: bacteria, fungi, long-term fertilization, Mesiq pro-sequencing, qPCR, black soil

王慧颖，徐明岗，周宝库，马想，段英华. 黑土细菌及真菌群落对长期施肥响应的差异及其驱动因素[J]. 中国农业科学, 2018, 51(5): 914-925.

WANG HuiYing, XU MingGang, ZHOU BaoKu, MA Xiang, DUAN YingHua. Response and Driving Factors of Bacterial and Fungal Community to Long-Term Fertilization in Black Soil[J]. Scientia Agricultura Sinica, 2018, 51(5): 914-925.

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