氮高效转化菌筛选及其堆肥应用

doi:10.11924/j.issn.1000-6850.casb2024-0240

中国农学通报 ›› 2024, Vol. 40 ›› Issue (32): 99-106.doi: 10.11924/j.issn.1000-6850.casb2024-0240

• 资源·环境·生态·土壤·气象 • 上一篇下一篇

氮高效转化菌筛选及其堆肥应用

许馨月(), 代文龙, 付海燕, 刘春光, 宋新宇, 杨峰山()

黑龙江大学，生命科学学院，农业微生物技术教育部工程研究中心，黑龙江省寒地生态修复与资源利用重点实验室，黑龙江省普通高校分子生物学重点实验室，哈尔滨 150080

收稿日期:2024-04-10 修回日期:2024-06-15 出版日期:2024-11-15 发布日期:2024-11-12
通讯作者:
杨峰山，男，1973年出生，山东文登人，教授，博士，主要从事作物害虫防治和农田土壤修复方面的研究。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院，Tel：0451-86608586，E-mail：yangfengshan@hlju.edu.cn。
作者简介:
许馨月，女，1999年出生，辽宁丹东人，在读硕士，研究方向：修复生态学。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院，Tel：0451-86608586，E-mail：15545180016@163.com。
基金资助:
黑龙江省“双一流”学科协同创新成果孵化项目“农业废弃物特种高效有机肥研制及配套大豆减障提升技术集成与示范”(LJGXCG2023-036); 黑龙江省留学回国创业基金“农业有机废弃物微生物转化菌剂研发与应用”(黑人社函〔2020〕381号)

Screening and Composting Application of Nitrogen-efficient Transformation Bacteria

XU Xinyue(), DAI Wenlong, FU Haiyan, LIU Chunguang, SONG Xinyu, YANG Fengshan()

Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Ecological Restoration and Resource Utilization for Cold Region & Key Laboratory of Molecular Biology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2024-04-10 Revised:2024-06-15 Published:2024-11-15 Online:2024-11-12

摘要/Abstract

摘要：

旨在筛选氮高效转化菌株并验证其在鸡粪堆肥过程中的效果，为国内畜禽粪污资源化利用与发展提供菌种资源和理论支持。以采集自不同区域的鸡粪和土壤样品为试材，用平板涂布法分离纯化菌株，采用氨氮纳氏比色法和紫外分光光度法检测菌株氮转化能力，并制备微生物菌剂应用于堆肥反应。结果表明：从样品中分离筛选出1株氮高效转化菌，鉴定为布鲁氏杆菌(Brucella tritici NC14)。选择生物炭作为最佳载体制备微生物菌剂并进行堆肥应用。在鸡粪堆肥试验中，施用微生物菌剂能够有效抑制42.3% NH₃的释放，减少NH₄⁺-N含量39.78%并增加NO₃^--N含量52.3%。研究表明，菌株NC14具有氮高效转化能力且有良好的应用潜力。

关键词: 氮转化, 鸡粪堆肥, 微生物菌剂, 异养硝化, 菌株筛选

Abstract:

The aim of this study is to screen nitrogen efficient transformation strains and verify their effects in the process of chicken manure composting, and to provide strain resources and theoretical support for the utilization and development of livestock and poultry manure in China. In this experiment, chicken manure and soil samples collected from different areas were used as the test materials, and the strains were isolated and purified by spread plate method. The nitrogen transformation ability of the strain was detected by Nessler's reagent colorimetric method and ultraviolet spectrophotometry. Then the microbial agent was prepared and applied to composting reaction. The results showed that 1 nitrogen efficient transformation bacterium was isolated and identified as Brucella tritici NC14. Biochar was selected as the best carrier to prepare microbial agent and then applied to compost. In the experiment of chicken manure composting, the application of microbial agent can effectively inhibit the release of NH₃ by 42.3%, reduce the content of NH₄⁺-N by 39.78% and increase the content of NO₃^--N by 52.3%. The results showed that strain NC14 had high nitrogen transformation ability and good application prospect.

Key words: nitrogen transformation, chicken manure compost, microbial agent, heterotrophic nitrification, screening

许馨月, 代文龙, 付海燕, 刘春光, 宋新宇, 杨峰山. 氮高效转化菌筛选及其堆肥应用[J]. 中国农学通报, 2024, 40(32): 99-106.

XU Xinyue, DAI Wenlong, FU Haiyan, LIU Chunguang, SONG Xinyu, YANG Fengshan. Screening and Composting Application of Nitrogen-efficient Transformation Bacteria[J]. Chinese Agricultural Science Bulletin, 2024, 40(32): 99-106.

图/表 9

时间/h	氨氮降解率/%
时间/h	NC25	JC12	HC42	NC14
24	57.12±1.03b	47.79±0.45c	43.71±0.99d	67.22±0.91a
48	57.49±0.80b	49.53±0.34c	46.54±0.61d	70.43±0.91a
72	59.52±0.99b	49.71±0.33c	47.72±1.04d	71.78±0.78a

表1. 氨氮降解率

时间/h	硝态氮含量/(mg/L)
时间/h	NC25	JC12	HC42	NC14
24	1.69±1.07b	1.71±0.96c	1.30±1.18c	3.21±0.86a
48	1.78±0.89c	1.79±0.89c	1.87±1.66b	3.95±1.39a
72	1.83±1.49c	1.81±2.25c	1.92±1.06b	4.20±1.09a

表2. 硝态氮含量

图1. 氮高效转化菌株NC14形态学鉴定

生理生化反应	NC14	生理生化反应	NC14
硝酸盐还原	+	V-P试验	-
氧化酶	+	葡萄糖分解	+
硫化氢	-	接触酶	+
甲基红试验	-	尿素	+
吲哚试验	-	柠檬酸盐利用	-

表3. 氮高效转化菌株NC14的生理生化反应特征

图2. 氮高效转化菌株NC14系统发育树

图3. 氮高效转化菌株NC14氮循环基因PCR产物琼脂糖凝胶电泳检测 a：amoA基因；b：hao基因；c：narG基因；d：nirS基因；e：nifH基因

图4. 氮高效转化菌株NC14硝化作用酶活测定不同小写字母表示处理间差异显著(P<0.05)

图5. 堆肥pH和氨气累积排放量变化 a：不同处理组pH变化；b：不同处理组NH3变化；每个值代表平均值±标准差(n=3)

图6. 铵态氮与硝态氮含量变化 a：不同处理组NH4+-N变化；b：不同处理组NO3--N变化；每个值代表平均值±标准差(n=3)

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氮高效转化菌筛选及其堆肥应用

Screening and Composting Application of Nitrogen-efficient Transformation Bacteria

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