盐碱胁迫对灰漠土氨氧化微生物丰度和群落多样性的影响

doi:10.11924/j.issn.1000-6850.casb2025-0263

摘要/Abstract

摘要：

新疆干旱区盐碱地面积大、分布广，其特殊的理化属性导致土壤肥力状况低下，且盐基离子会干扰氮素转化。目前，盐碱环境对氨氧化细菌等硝化关键微生物的影响尚不明确，制约了对该区域氮素转化机制的深入研究及盐碱地改良工作的开展。本研究通过土柱模拟试验，旨在探究NaCl和Na₂CO₃+NaHCO₃胁迫对干旱区灰漠土土壤氨氧化古菌(AOA)和氨氧化细菌(AOB)群落多样性的影响。试验设置了非盐（碱）化(CK)、中度盐化(CS)和中度碱化(AS) 3个处理，评估了盐碱胁迫对棉花生长、土壤理化性质以及AOA和AOB的丰度和群落结构的影响。结果表明：盐碱胁迫显著降低了棉花株高和干物质重、土壤NO₃-N含量和潜在硝化速率(PNR)；然而，土壤含水量、盐分、pH和NH₄-N含量显著增加。与对照组相比，盐碱胁迫处理AOA和AOB的基因拷贝数均显著降低，且AOA/AOB比值也显著降低。此外，在所有处理下，AOB的基因拷贝数均显著高于AOA。PNR与AOA丰度和AOB丰度均呈显著正相关关系。不同处理下AOB群落操作分类单元(OTUs)的数量均显著高于AOA，碱胁迫显著降低了AOB群落的OTU数量，而盐胁迫显著降低了AOB群落的辛普森指数和香浓指数，碱胁迫显著增加了AOB群落的辛普森指数。盐胁迫显著降低了AOA和AOB的Pielou_e均匀度指数，碱胁迫显著降低了AOA和AOB的Chao1丰富度指数，但是显著增加了其Pielou_e均匀度指数。在门水平上，AOA群落的优势菌门为Thaumarchaeota；AOB群落的优势菌门为Proteobacteria；在属水平上，AOA优势菌属为Candidatus Nitrosocaldus和Nitrososphaera，AOB优势菌属为Nitrosospira和Nitrosomonas。盐碱胁迫显著增加了Candidatus Nitrosocaldus和Nitrosomonas的相对丰度，但是显著降低了Nitrososphaera和Nitrosospira的相对丰度。冗余分析显示：AOA群落结构的改变主要驱动因子为土壤含水量和pH，而AOB群落结构的改变主要驱动因子为pH。综上，AOA和AOB共同参与了灰漠土的硝化作用，土壤pH是影响其丰度、生长及群落结构的主导环境因子。

关键词: 盐碱胁迫, 氨氧化古菌, 氨氧化细菌, 潜在硝化势, 高通量测序

Abstract:

Saline-alkali soils are prevalent in the arid regions of Xinjiang, unique physical and chemical properties lead to low soil fertility and the base ions interfere with nitrogen transformation processes. Currently, the effects of the saline-alkali environment on key nitrifying microorganisms, such as ammonia-oxidizing bacteria, remain poorly understood, limiting research on nitrogen cycling in these soils and hindering efforts to improve saline-alkali land. This study aimed to evaluate the effects of two types of saline-alkali stress (NaCl and Na₂CO₃ + NaHCO₃) on the abundance, diversity, and community structure of ammonia-oxidizing microorganisms in gray desert soil from arid areas. This experiment used three treatments, including chloride stress (CS), alkaline stress (AS) and a control group (CK) with no salt-alkali stress, respectively, to evaluate the effects of saline-alkaline stress on cotton growth, soil physicochemical and the abundance and community structure of soil AOA and AOB. The results showed that saline-alkaline stress significantly reduced plant height and dry matter weight of cotton as well as soil NO₃-N content and potential nitrification rate (PNR). Soil moisture content, salinity, pH and NH₄-N content increased significantly under stress conditions. Both salt and alkaline stress decreased the amoA gene copy numbers of AOA, AOB, and the AOA/AOB ratios, respectively, with AOB showing higher amoA gene copy numbers than that of AOA. PNR was positively correlated with AOA and AOB amoA gene copy numbers. Additionally, the operational taxonomic units (OTUs) of AOB were more abundant than those of AOA under saline alkaline stress with alkaline stress causing a significant decrease in OTUs of AOB. Salt stress significantly decreased the Simpson and Shannon diversity index of AOB, whereas alkaline stress significantly increased the Simpson index of AOB. Salt stress also decreased the Pielou_e index of both AOA and AOB, while, alkaline stress increased the Chao1 index of AOA and AOB. The dominant phylum of the AOA community was Thaumarchaeota, while, Proteobacteria was the dominant phylum in the AOB community. The dominant genera for AOA were Candidatus Nitrosocaldus and Nitrososphaera, and they were Nitrosospira and Nitrosomonas for AOB. Both salt and alkaline stress significantly increased the relative abundance of Candidatus Nitrosocosmicus, Nitrosomonas, while decreasing Nitrososphaera, Nitrosospira. Redundancy analysis showed that soil properties, including SWC and pH played an important role in shaping the AOA and denitrifier communities, while AOB community structure was only significantly correlated with pH. These findings suggest that AOA and AOB contribute to nitrification in alluvial gray desert soils, with pH being the dominant factor affecting the growth of ammonia-oxidizing microorganisms and community structure.

Key words: saline-alkaline stress, AOA, AOB, potential nitrification rate, high-throughput sequencing

向贵琴, 叶扬, 张楠, 郭慧娟. 盐碱胁迫对灰漠土氨氧化微生物丰度和群落多样性的影响[J]. 中国农学通报, 2025, 41(30): 79-89.

XIANG Guiqin, YE Yang, ZHANG Nan, GUO Huijuan. Effects of Saline and Alkaline Stress on Abundance and Diversity of Community of Ammonia Oxidizing Microorganisms in Gray Desert Soil[J]. Chinese Agricultural Science Bulletin, 2025, 41(30): 79-89.

图/表 12

参考文献 54

[1]	SUN H, SHI W, ZHOU M, et al. Effect of biochar on nitrogen use efficiency, grain yield and amino acid content of wheat cultivated on saline soil[J]. Plant, soil and environment, 2019, 65(2):83-89.
[2]	杨劲松, 姚荣江, 王相平, 等. 防止土壤盐渍化,提高土壤生产力[J]. 科学, 2021, 73(6):2,4,30-34.
[3]	JACOBSEN C S, HELMS H M. Agricultural soils, pesticides and microbial diversity[J]. Current opinion in biotechnology, 2014, 27:15-20. doi: 10.1016/j.copbio.2013.09.003 pmid: 24863892
[4]	VAN DER HEIJDEN M G, BRUIN S D, LUCKERHOFF L, et al. A widespread plant-fungal-bacterial symbiosis promotes plant biodiversity, plant nutrition and seedling recruitment[J]. The ISME journal, 2016, 10(2):389-399.
[5]	CHEN Z, ZHOU T, HUANG G, et al. Soil microbial community and associated functions response to salt stresses: Resistance and resilience[J]. Science of the total environment, 2024, 954:176475.
[6]	高敬文, 郭子燕, 王峰. 盐碱胁迫下作物氮素吸收利用的响应机理及调控措施研究进展[J]. 中国农学通报, 2024, 40(24):44-50. doi: 10.11924/j.issn.1000-6850.casb2023-0828
[7]	ZHAO X, GAO J, YU X, et al. Evaluation of the microbial community in various saline alkaline-soils driven by soil factors of the Hetao Plain, Inner Mongolia[J]. Scientific reports, 2024, 14(1):28931.
[8]	ZHOU Y, HE Z, LIN Q, et al. Salt stress affects the bacterial communities in rhizosphere soil of rice[J]. Frontiers in microbiology, 2024, 15:1505368.
[9]	PAN H, LIU H, LIU Y, et al. Understanding the relationships between grazing intensity and the distribution of nitrifying communities in grassland soils[J]. Science of the total environment, 2018, 634:1157-1164.
[10]	LOUCA S, DOEBELI M. Taxonomic variability and functional stability in microbial communities infected by phages[J]. Environmental microbiology, 2017, 19(10):3863-3878. doi: 10.1111/1462-2920.13743 pmid: 28371143
[11]	李红强, 姚荣江, 杨劲松, 等. 盐渍化对农田氮素转化过程的影响机制和增效调控途径[J]. 应用生态学报, 2020, 31(11):3915-3924. doi: 10.13287/j.1001-9332.202011.023
[12]	杨易, 黄立华, 肖扬, 等. 苏打盐碱化稻田土壤氮素矿化和硝化特征及其影响因子[J]. 植物营养与肥料学报, 2022, 28(10):1816-1827.
[13]	CUI G, ZHANG Y, ZHANG W, et al. Response of carbon and nitrogen metabolism and secondary metabolites to drought stress and salt stress in plants[J]. Journal of plant biology, 2019, 62:387-399.
[14]	HU M, LE Y, SARDANS J, et al. Moderate salinity improves the availability of soil P by regulating P-cycling microbial communities in coastal wetlands[J]. Global change biology, 2023, 29(1):276-288.
[15]	YANG C, CHENY, SUN W, et al. Extreme soil salinity reduces N and P metabolism and related microbial network complexity and community immigration rate[J]. Environmental research, 2025, 264:120361.
[16]	李亚威, 徐俊增, 卫琦, 等. 不同水盐条件下盐渍土硝化过程特征[J]. 排灌机械工程学报, 2018, 36(9): 909-913.
[17]	PATIL P K, BASKARAN V, VINAY T N, et al. Abundance, community structure and diversity of nitrifying bacterial enrichments from low and high saline brackish water environments[J]. Letters in applied microbiology, 2021, 73(1):96-106.
[18]	ZHANG S Y, ZHANG H F, LIU H M, et al. Combined organic and inorganic fertilization increases soil network complexity and multifunctionality in a 5- year fertilization system[J]. Land degradation and development, 2024, 35(2):586-596.
[19]	KUYPERS M M, MARCHANT H K, KARTAL B. The microbial nitrogen-cycling network[J]. Nature reviews microbiology, 2018, 16(5):263-276. doi: 10.1038/nrmicro.2018.9 pmid: 29398704
[20]	HÖRSTMANN C, RAES E J, BUTTIGIEG P L, et al. Hydrographic fronts shape productivity, nitrogen fixation, and microbial community composition in the southern Indian Ocean and the Southern Ocean[J]. Biogeosciences, 2021, 18(12):3733-3749.
[21]	HUANG D, LI X, LUO X. Response of nitrifier and denitrifier abundance to salinity gradients in agricultural soils at the Yellow River Estuary[J]. Agronomy, 2022, 12(7):1642.
[22]	CUI Y W, ZHANG H Y, DING J R, et al. The effects of salinity on nitrification using halophilic nitrifiers in a Sequencing Batch Reactor treating hypersaline wastewater[J]. Scientific reports, 2016, 6:24825.
[23]	ZHANG Y, CHEN L, DAI T, et al. The influence of salinity on the abundance, transcriptional activity, and diversity of AOA and AOB in an estuarine sediment: a microcosm study[J]. Applied microbiology and biotechnology, 2015, 99(22):9825-9833. doi: 10.1007/s00253-015-6804-x pmid: 26219499
[24]	ZOU D, CHEN J, ZHANG C, et al. Diversity and salinity adaptations of ammonia oxidizing archaea in three estuaries of China[J]. Applied microbiology and biotechnology, 2023, 107(22):6897-6909. doi: 10.1007/s00253-023-12761-4 pmid: 37702790
[25]	鲍士旦. 土壤农化分析[M]. 北京: 中国农业出版社, 1981.
[26]	KUROLA J, SALKINOJA-SALONEN M, AARNIO T, et al. Activity, diversity and population size of ammonia-oxidising bacteria in oil-contaminated landfarming soil[J]. FEMS microbiology letters, 2005, 250(1):33-38. pmid: 16043309
[27]	HU H W, ZHANG L M, DAI Y, et al. pH-dependent distribution of soil ammonia oxidizers across a large geographical scale as revealed by high-throughput pyrosequencing[J]. Journal of soils and sediments, 2013, 13(8):1439-1449.
[28]	EBIE Y, NODA N, MIURA H, et al. Comparative analysis of genetic diversity and expression of amoA in wastewater treatment processes[J]. Applied microbiology and biotechnology, 2004, 64(5):740-744. pmid: 14758520
[29]	马丽娟, 张慧敏, 侯振安, 等. 长期咸水滴灌对土壤氨氧化微生物丰度和群落结构的影响[J]. 农业环境科学学报, 2019, 38(12):2797-2807.
[30]	陈竹, 郭岩彬, 孟凡乔, 等. 施肥方式对干旱半干旱地区土壤氨氧化微生物数量和群落的影响[J]. 生态学报, 2021, 41(11):4577-4585.
[31]	LIANG J, SHI W. Cotton/halophytes intercropping decreases salt accumulation and improves soil physicochemical properties and crop productivity in saline-alkali soils under mulched drip irrigation: a three-year field experiment[J]. Field crops research, 2021, 262:108027.
[32]	肖雯丽, 王含瑞, 王梦亮, 等. 盐碱胁迫下植物响应机制的研究进展[J]. 中国农学通报, 2024, 40(33):78-85. doi: 10.11924/j.issn.1000-6850.casb2023-0775
[33]	郭家鑫, 鲁晓宇, 陶一凡, 等. 盐碱胁迫对棉花生长和养分吸收的影响[J]. 干旱地区农业研究, 2022, 40(4):23-32,59.
[34]	AMINI S, GHADIRI H, CHEN C R, et al. Salt-affected soils, reclamation, carbon dynamics, and biochar: a review[J]. Journal of soils and sediments, 2016, 16:939-953.
[35]	MA T, ZENG W, LI Q, et al. Effects of water, salt and nitrogen stress on sunflower (Helianthus annuus L.) at different growth stages[J]. Journal of soil science and plant nutrition, 2016, 16(4):1024-1037.
[36]	BERNHARD A E, BOLLMANN A. Estuarine nitrifiers: new players, patterns and processes[J]. Estuarine, coastal and shelf science, 2010, 88(1):1-11.
[37]	HE H, ZHEN Y, MI T, et al. Ammonia-oxidizing Archaea and Bacteria differentially contribute to ammonia oxidation in sediments from adjacent waters of Rushan Bay, China[J]. Frontiers in microbiology, 2018, 9:116. doi: 10.3389/fmicb.2018.00116 pmid: 29456526
[38]	AKHTAR M, HUSSAIN F, ASHRAF M Y, et al. Influence of salinity on nitrogen transformations in soil[J]. Communications in soil science and plant analysis, 2012, 43(12):1674-1683.
[39]	HOU L, ZHENG Y, LIU M, et al. Anaerobic ammonium oxidation (anammox) bacterial diversity, abundance, and activity in marsh sediments of the Yangtze Estuary[J]. Journal of geophysical research: biogeosciences, 2013, 118(3):1237-1246.
[40]	DENG F Y, HOU L J, LIU M, et al. Dissimilatory nitrate reduction processes and associated contribution to nitrogen removal in sediments of the Yangtze Estuary[J]. Journal of geophysical research: biogeosciences, 2015, 120(8):1521-1531.
[41]	SONG L, WANG J, PAN J, et al. Chronic nitrogen enrichment decreases soil gross nitrogen mineralization by acidification in topsoil but by carbon limitation in subsoil[J]. Geoderma, 2022, 428:116159.
[42]	KESHRI J, MISHRA A, JHA B. Microbial population index and community structure in saline-alkaline soil using gene targeted metagenomics[J]. Microbiological research, 2013, 168:165-173. doi: 10.1016/j.micres.2012.09.005 pmid: 23083746
[43]	MAGALHÃES C M, MACHADO A, BORDAL A A. Temporal variability in the abundance of ammonia-oxidizing bacteria vs.archaea in sandy sediments of the Douro River estuary, Portugal[J]. Aquatic microbial ecology, 2009, 56(1):13-23.
[44]	ZHANG L M, HU H W, SHEN J P, et al. Ammonia-oxidizing archaea have more important role than ammonia-oxidizing bacteria in ammonia oxidation of strongly acidic soils[J]. The ISME journal, 2012, 6(5):1032-1045.
[45]	SONG W, LIU J, QIN W, et al. Functional traits resolve mechanisms governing the assembly and distribution of nitrogen-cycling microbial communities in the global ocean[J]. Mbio, 2022, 13(2):e03832-21.
[46]	WAKELIN S, WILLAMS E, O'SULLIVAN C A, et al. Predicting the efficacy of the nitrification inhibitor dicyandiamide in pastoral soils[J]. Plant and soil, 2014, 381:35-43.
[47]	李晨华, 贾仲君, 唐立松, 等. 不同施肥模式对绿洲农田土壤微生物群落丰度与酶活性的影响[J]. 土壤学报, 2012, 49(3):567-574.
[48]	何园, 胡文革, 马得草, 等. 艾比湖湿地盐节木根际土壤氨氧化微生物多样性和丰度及其与环境因子的相关性分析[J]. 环境科学学报, 2017, 37(5):1967-1975.
[49]	ZHANG Z, FENG S, LUO J, et al. Evaluation of microbial assemblages in various saline-alkaline soils driven by soluble salt ion components[J]. Journal of agricultural and food chemistry, 2021, 69(11):3390-3400. doi: 10.1021/acs.jafc.1c00210 pmid: 33703896
[50]	CANFORA L, BACCI G, PINZARI F, et al. Salinity and bacterial diversity: to what extent does the concentration of salt affect the bacterial community in a saline soil[J]. Plos one, 2014, 9:0106662
[51]	GAO L, RAO M P N, LIU Y H, et al. SALINITY-Induced changes in diversity, stability, and functional profiles of microbial communities in different Saline Lakes in Arid Areas[J]. Microbial ecology, 2024, 87(1):135. doi: 10.1007/s00248-024-02442-8 pmid: 39482450
[52]	NI G, LEUNG P M, DAEBELER A, et al. Nitrification in acidic and alkaline environments[J]. Essays in biochemistry, 2023, 67(4):753-768. doi: 10.1042/EBC20220194 pmid: 37449414
[53]	CAMPBELL B J, KIRCHMAN D L. Bacterial diversity, community structure and potential growth rates along an estuarine salinity gradient[J]. The ISME Journal, 2013, 7:210-220.
[54]	ZHENG W, XUE D, LI X, et al. The responses and adaptations of microbial communities to salinity in farmland soils: a molecular ecological network analysis[J]. Applied soil ecology, 2017, 120:239-246.

处理	盐碱类型及盐碱化程度	电导率EC_1:5/(dS/m)	pH(1:2.5)
CK	非盐（碱）化土壤	0.18	8.17
CS	NaCl中度盐化土壤	1.41	8.41
AS	Na₂CO₃+NaHCO₃中度碱化土壤	0.65	9.90

处理	盐碱类型及盐碱化程度	电导率EC_1:5/(dS/m)	pH(1:2.5)
CK	非盐（碱）化土壤	0.18	8.17
CS	NaCl中度盐化土壤	1.41	8.41
AS	Na₂CO₃+NaHCO₃中度碱化土壤	0.65	9.90

处理	土壤含水量/%	电导率EC/(ds/m)	pH	全氮TN/(g/kg)	总碳TC/(g/kg)	铵态氮NH₄-N/(mg/kg)	硝态氮NO₃-N/(mg/kg)
CK	11.88±1.472c	0.33±0.067c	8.15±0.046c	0.59±0.036b	14.75±0.621b	7.55±0.415c	145.30±1.486a
CS	17.18±1.814a	1.56±0.115a	8.49±0.035b	0.58±0.021b	14.44±0.191b	14.91±0.663b	133.55±1.440b
AS	14.0±1.4402b	0.66±0.010b	9.79±0.152a	0.69±0.021a	16.99±0.995a	23.75±0.524a	121.96±1.842c

处理	土壤含水量/%	电导率EC/(ds/m)	pH	全氮TN/(g/kg)	总碳TC/(g/kg)	铵态氮NH₄-N/(mg/kg)	硝态氮NO₃-N/(mg/kg)
CK	11.88±1.472c	0.33±0.067c	8.15±0.046c	0.59±0.036b	14.75±0.621b	7.55±0.415c	145.30±1.486a
CS	17.18±1.814a	1.56±0.115a	8.49±0.035b	0.58±0.021b	14.44±0.191b	14.91±0.663b	133.55±1.440b
AS	14.0±1.4402b	0.66±0.010b	9.79±0.152a	0.69±0.021a	16.99±0.995a	23.75±0.524a	121.96±1.842c

处理	AOA基因拷贝数/(×10⁶ copies/g 干土)	AOB基因拷贝数/(×10⁷ copies/g 干土)	AOA/AOB
CK	1.45±0.406a	5.32±0.717a	0.0273±0.002a
CS	0.36±0.079b	4.36±0.759b	0.0084±0.001c
AS	0.43±0.73b	3.83±0.558c	0.0113±0.001b