硝化抑制剂和肥料减施对减少设施菜田氮源气体排放及与微生物功能基因的互作

doi:10.7668/hbnxb.20191952

摘要/Abstract

摘要： 开展了以减少设施菜田系统中氮源气体损失、提高肥料利用率为目的的田间试验。以设施黄瓜为例，通过设置对比常规施肥、减量施肥与不同硝化抑制剂型配比试验，连续5 a定位监测不同施肥处理下氮源气体排放规律，基于qPCR结果分析了氮源气体排放与氨氧化古菌AOA的互作关系，并应用高通量Illumina MiSeq平台测序，利用PICRUSt对16S rDNAV3-V4区基因系列数据进行分析，基于KEGG数据库对与氮代谢相关的功能基因进行了鉴定。结果表明，北方设施黄瓜-土壤系统的5个不同施肥处理中，增施硝化抑制剂DCD（处理In-1）与常规施肥（处理FT）和减量施肥（处理OPT）相比，每年向环境排放的氮氧化物（NO_x）排放总量分别减少76.5%和71.8%，减量施肥（处理OPT）和增施DCD分别比常规施肥（处理FT）氧化亚氮（N₂O）年排放总量减少28.4%和21.8%。明确了北方石灰性褐土设施黄瓜菜田中，增施DCD可显著抑制氨氧化古菌AOA种群数量，效果优于吡啶；氨氧化古菌AOA对NO_x排放起调控作用，氨氧化古菌AOA amoA基因拷贝数与土壤NO_x排放通量之间呈指数正相关关系：Y=121.18e^3E-08x（R²=0.840 5，P<0.05）。基于KEGG数据库的鉴定结果显示，在肥料减施（处理OPT）和硝化抑制剂（处理In-2）调控的土壤中，与同化途径相关的烯酰水合酶（K01692）、底物结合蛋白（K02035）以及转运系统ATP结合蛋白（K09687）系统代谢通路相关的功能基因相对丰度高，是响应上述调控变化最为敏感的氮循环途径。5 a间，与常规施肥（处理FT）相比，减量施肥（处理OPT）和增施DCD（处理In-1）的肥料偏生产力分别提升了32.1%~38.2%和17.8%~21.9%。因此，在北方设施菜田中，增施施氮量15%的硝化抑制剂和减量25%的肥料施用，可有效减少农田氮源气体排放损失，提高肥料利用率，并且明确了随着氨氧化古菌AOA种群数量的增加，NO_x的排放通量呈指数锐升的关系。为实现农业面源污染有效防控，促进我国农业高质量绿色发展提供了有力的科学依据。

关键词: 设施菜田, 氮源气体减排, 氨氧化古菌, 硝化抑制剂, 微生物功能基因

Abstract: It was carried out this experiment to reduce nitrogen source gas emission and improve fertilizer efficiency in greenhouse vegetable field. By setting up a comparison among conventional fertilization of farmers pattern, fertilizer reduction and different nitrification inhibitors in the greenhouse cucumber soil, the gas emission regularity of nitrogen source and yields were monitored with different fertilization treatments for five years. Based on qPCR when compared with conventional fertilization of farmers treatment (FT) and fertilizer reduction treatment (OPT). The total annual emission of nitrous oxide (N₂O) reduced 28.4% and 21.8% respectively by fertilizer reduction (OPT) and applying nitrification inhibitor DCD (In-1) compared with conventional fertilization (FT).The interaction between nitrogen oxides (NO_x) emission and ammonia-oxidizing archaea was analyzed. On the high-throughput sequencing Illumina MiSeq platform, metagenomes were predicted from 16S rDNA gene sequence data using PICRUSt, and functional genes associated with nitrogen metabolisms were identified based on KEGG database. The results showed that the total annual emission of nitrogen oxides (NO_x) by applying nitrification inhibitor DCD (treatment In-1) was respectively reduced by 76.5% and 71.8%. It was found that DCD (In-1) significantly inhibited Ammonia-oxidizing archaea (AOA) population, and the effect was better than pyridine (In-2) in calcareous cinnamon soil in the North greenhouse. There was a positive exponential correlation between the gene copy numbers of AOA and soil NO_x flux. The equation was as follows:Y=121.18e^3E-08x (R ²=0.840 5, P<0.05). The identification results based on KEGG database showed that the relative abundance of enoyl-CoA hydratase(K01692), substrate-binding protein(K02035), transport system ATP-binding protein(K09687) functional genes associated with assimilatory pathways was higher than others. The three functional genes metabolic pathways in soil were the most sensitive nitrogen cycle pathways responding to the variation with the regulation of nitrification inhibitors and fertilizer reduction. In five years,compared with conventional fertilization of farmers' treatment (FT), the partial factor productivity of fertilizer with fertilizer reduction treatment (OPT) and DCD treatment (In-1) increased by 32.1%-38.2% and 17.8%-21.9% respectively. It was suggested that application of nitrification inhibitors (weight of 15% N) and fertilizer reduction (25%) effectively reduced nitrogen source gas emission and improved fertilizer utilization rate in the North greenhouse cucumber soil. It was found that the emission flux of NO_x increased sharply with the increase of AOA population. This study provided a strong scientific basis for the effective prevention and control of agricultural non-point source pollution and the promotion of high quality and green development of agriculture.

Key words: Greenhouse vegetable soil, Reducing nitrogen emission, Ammonia-oxidizing archaea, Nitrification inhibitor, Microbial functional genes

中图分类号:

S154.36
S19

王凌, 张国印, 刘孟朝, 孙世友, 刘蕾, 茹淑华, 赵欧亚, 李博文. 硝化抑制剂和肥料减施对减少设施菜田氮源气体排放及与微生物功能基因的互作[J]. 华北农学报, 2021, 36(2): 196-203. doi: 10.7668/hbnxb.20191952.

WANG Ling, ZHANG Guoyin, LIU Mengchao, SUN Shiyou, LIU Lei, RU Shuhua, ZHAO Ouya, LI Bowen. Effect of Nitrification Inhibitor and Fertilizer Reduction on Reducing Nitrogen Source Gas Emission and Their Interaction with Microbial Functional Genes in Greenhouse Vegetable Field[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(2): 196-203. doi: 10.7668/hbnxb.20191952.

http://www.hbnxb.net/CN/Y2021/V36/I2/196

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