基于高通量绝对定量对施用不同改良物料后盐碱地土壤细菌群落影响的研究

Bacterial Community in Saline-alkali Soil after Application of Different Modified Materials Based on High-Throughput Absolute Quantification

  • 摘要:
    目的 采用高通量绝对定量方法探究改良物料对盐碱土壤细菌群落结构的影响,可弥补相对定量在研究盐碱土壤细菌群落变化方面的不足。
    方法 采用大田试验,以玉米为供试作物,设置对照(CK)、脱硫石膏(T1, 15 t hm−2)、土壤改良剂(T2, 15 t hm−2)、有机肥(T3, 7.5 t hm−2)、脱硫石膏(T4, 7.5 t hm−2 + 土壤改良剂7.5 t hm−2 + 有机肥3.75 t hm−2)5个处理,在玉米种植84天后利用高通量绝对定量技术测定土壤细菌绝对丰度、多样性和群落结构的变化。
    结果 施用改良物料可以降低土壤pH和盐分含量,其中,T4处理对降低pH的效果最显著,相比CK下降了0.44个单位(P < 0.05)。施用改良物料提高了土壤速效磷、速效钾、有机质和碱解氮含量,其中T4处理的速效磷、速效钾、有机质和碱解氮含量较CK分别提高59%、41.59%、25.44%和47.51%(P < 0.05)。绝对定量分析结果表明,变形菌门、酸杆菌门、拟杆菌门、芽单胞菌门、放线菌门和绿弯菌门是细菌优势菌门,施用改良物料增加了土壤细菌群落的丰富度和多样性,细菌16S rRNA基因拷贝数显著高于CK,且T4处理在增加细菌16S rRNA基因拷贝数方面好于其余改良处理。相关性分析结果表明土壤细菌优势类群(门水平和属水平)与速效磷、速效钾、有机质和碱解氮呈正相关或显著正相关,而与pH和盐分含量呈负相关。
    结论 施用改良物料降低了盐碱土壤pH和盐分含量,提高了土壤养分含量、细菌多样性和绝对数量。同时,引入高通量绝对定量方法开展研究能够弥补相对丰度在表征微生物群落方面的缺陷,因而在阐明微生态学许多悬而未决的问题具有一定潜力,可将这一技术引入土壤微生物群落的研究中。

     

    Abstract:
    Objective The high-throughput absolute quantitative method was used to explore the effect of modified materials on the bacterial community structure of saline-alkali soil, which could make up for the deficiency of relative quantification in studying the changes of bacterial community in saline-alkali soil.
    Method A field experiment was conducted with maize as the test crop, five treatments were set up: CK (blank control), T1 (desulfurization gypsum 15 t hm−2), T2 (soil amendment 15 t hm−2), T3 (organic fertilizer 7.5 t hm−2) and T4 (desulfurization gypsum 7.5 t hm−2 + soil amendment 7.5 t hm−2 + organic fertilizer 3.75 t hm−2). After 84 days of maize planting, the absolute abundance, diversity and community structure of soil bacteria were determined by high-throughput absolute quantitative technique.
    Result The application of modified materials could reduce soil pH and salt content. Among them, T4 treatment had the most significant effect on reducing pH, which was 0.44 units lower than CK (P < 0.05). The application of modified materials increased the contents of soil available phosphorus (AP), available potassium (AK), soil organic matter (SOM) and alkaline hydrolysis nitrogen (AN). The contents of AP, AK, SOM and AN in T4 treatment increased by 59%, 41.59%, 25.44% and 47.51%, respectively, compared with CK (P < 0.05). The results of absolute quantitative analysis showed that Proteobacteria, Acidobacteria, Bacteroidetes, Gemmatimonadetes, Actinobacteria and Chloroflexi were the dominant bacteria. The application of modified materials increased the richness and diversity of soil bacterial community. The copy number of bacterial 16S rRNA gene was significantly higher than that of CK, and T4 treatment was better than other modified treatments in increasing the copy number of bacterial 16S rRNA gene. The results of correlation analysis showed that the dominant groups of soil bacteria (phylum level and genus level) were positively or significantly positively correlated with AP, AK, SOM and AN, but negatively correlated with pH and salt content.
    Conclusion The application of modified materials reduced the pH and salt content of saline-alkali soil, and increased the soil nutrient content, bacterial diversity and absolute number. The introduction of high-throughput absolute quantitative methods to carry out research can make up for the defects of relative abundance in characterizing microbial communities. Therefore, it has certain potential in elucidating many outstanding problems in micro-ecology, and this technology can be introduced into the study of soil microbial communities.

     

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