2株可培养细菌氮代谢特性的研究

doi:10.11924/j.issn.1000-6850.casb20191100789

中国农学通报 ›› 2020, Vol. 36 ›› Issue (30): 106-111.doi: 10.11924/j.issn.1000-6850.casb20191100789

2株可培养细菌氮代谢特性的研究

刘志新¹^,²(), 姚娜², 刘灵芝²()

¹北大荒垦丰种业股份有限公司曙光农场分公司,黑龙江佳木斯 154451
²沈阳农业大学土地与环境学院,沈阳 110161

收稿日期:2019-11-03 修回日期:2019-11-26 出版日期:2020-10-25 发布日期:2020-10-16
通讯作者: 刘灵芝
作者简介:刘志新,男,1972年出生,黑龙江桦南人,农业技术员,学士,研究方向：土壤肥力与农业生产。通信地址：154454 黑龙江省桦南县曙光农场北大荒垦丰种业股份有限公司,Tel：18804003669,E-mail： 64066604@qq.com。
基金资助:
国家自然科学基金“冷凉地区苹果园土壤氨氧化微生物种群结构与氮循环关系研究”(31101504);辽宁省高等学校基本科研项目“氮代谢功能基因表达调控施肥土壤氮素转化的作用机制”(LSNZD201705);辽宁省自然科学基金“硝化古菌响应秸秆还田土壤中氮素转化的作用机制”(2019-MS-271)

Nitrogen Metabolic Characteristics of Two Cultivable Bacteria

Liu Zhixin¹^,²(), Yao Na², Liu Lingzhi²()

¹Shuguang Subsidiary Company of Great Northern Wilderness Kenfeng Seed Co., Ltd., Jiamusi Heilongjiang 154451
²College of Land and Environment, Shenyang Agricultural University, Shenyang 110161

Received:2019-11-03 Revised:2019-11-26 Online:2020-10-25 Published:2020-10-16
Contact: Liu Lingzhi

摘要/Abstract

摘要：

研究旨在分析土壤中可培养细菌菌株的氮代谢特征,并进一步探讨微生物在土壤氮素转化中的可能作用机制。以2株分离自苹果园土壤的细菌菌株SY5-4和SY11-10为试材,采用传统培养方法结合分子检测技术,分别测定菌株生长特性及其氮素转化能力。研究结果表明,异养条件下,菌株SY5-4和SY11-10的世代时间分别为243.5 min和202.7 min。菌株生长过程中,培养液中铵态氮浓度始终维持在较高水平,铵态氮、亚硝态氮和硝态氮浓度均表现出先升后降的趋势。硝化（amoA和hao）和反硝化（nosZ、norB、nirK和nap）基因检测结果表明,菌株SY11-10具有多种氮素转化潜能。综上,供试菌株培养过程中,培养液中氮素发生变化,并在菌体中检测到不同氮转化基因,表明菌株参与多种氮代谢途径。

关键词: 可培养细菌, 异养培养基, 氮代谢, 硝化基因, 反硝化基因

Abstract:

To study the N metabolic characteristics of culturable bacteria in soil, and discuss the possible mechanism of microorganisms in soil N transformation, 2 strains, named SY5-4 and SY11-10 isolated from apple orchard soil were selected to analyze the growth characteristics, and their capabilities of nitrogen transformation, by using the traditional culture method combined with molecular technology. The results showed that, in heterotrophic condition, the generation time of SY5-4 and SY11-10 was 243.5 min and 202.7 min, respectively. The concentration of NH₄⁺-N in the medium maintained at a high level, while the concentration of NH₄⁺-N, NO₂^--N and NO₃^--N increased first and decreased afterwards, in the process of bacterial growth. The nitrification (amoA and hao), and denitrification (nosZ, norB, nirK and nap) gene detection showed that, strain SY11-10 had a variety of N transformation potential. In conclusion, the nitrogen in medium changed in the process of bacterial growth, and different nitrogen transformation genes were detected in the tested strains, which indicated that the isolated strains participated in various N metabolism pathways.

Key words: culturable bacteria, heterotrophic medium, nitrogen metabolism, nitrification gene, denitrification gene

中图分类号:

S154.3

刘志新, 姚娜, 刘灵芝. 2株可培养细菌氮代谢特性的研究[J]. 中国农学通报, 2020, 36(30): 106-111.

Liu Zhixin, Yao Na, Liu Lingzhi. Nitrogen Metabolic Characteristics of Two Cultivable Bacteria[J]. Chinese Agricultural Science Bulletin, 2020, 36(30): 106-111.

图/表 4

参考文献 30

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2株可培养细菌氮代谢特性的研究

Nitrogen Metabolic Characteristics of Two Cultivable Bacteria

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引物名称与序列(5’-3’)	参考文献
amoA-1F: 5’-GGGGTTTCTACTGGTGGT-3’ amoA-2R: 5’-CCCCTCKGSAAAGCCTTCTTC-3’	[23]
nosZ-1870F: 5’-GCRACGGCAASAAGGTSMSSGT-3’ nosZ-2090R: 5’-CAKRTGCAKSGCRTGGCAGAA-3’	[24]
norB-1F: 5’- CGNGARTTYCTSGARCARCC-3’ norB-8R: 5’-CRTADGCVCCRWAGAAVGC-3’	[25]
nirK-1F: 5’-GGMATGGTKCCSTGGCA-3’ nirK-3R: ’-GAACTTGCCGGTVGYCCAGAC-3’	[26]
nap-1F: 5’-TCTGGACCATGGGCTTCAACCA-3’ nap-1R: 5’-ACGACGACCGGCCAGCGCA-3’	[27]
hao-1F: 5’-TGCGTGGARTGYCAC-3’ hao-3R: 5’-AGRTARGAKYSGGCAAA-3’	[28]

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