2024年5月29日发(作者：长孙云天)

中国生态农业学报(中英文)

2020年10月 第28卷 第10期

Chinese Journal of Eco-Agriculture, Oct. 2020, 28(10): 16181629

DOI: 10.13930/.200160

孙建平, 刘雅辉, 左永梅, 韩民利, 张虹伟, 吕晶晶. 盐地碱蓬根际土壤细菌群落结构及其功能[J]. 中国生态农业学报

(中英文), 2020, 28(10):

16181629

SUN J P, LIU Y H, ZUO Y M, HAN M L, ZHANG H W, LYU J J. The bacterial community structure and function of Suaeda

salsa rhizosphere soil[J]. Chinese Journal of Eco-Agriculture, 2020, 28(10):

16181629

盐地碱蓬根际土壤细菌群落结构及其功能

孙建平, 刘雅辉

**

, 左永梅, 韩民利, 张虹伟, 吕晶晶

(河北省农林科学院滨海农业研究所/河北省盐碱地绿化工程技术中心/唐山市耐盐植物重点实验室 唐山 063299)

*

摘 要: 盐地碱蓬作为生物改良盐碱地的理想材料, 其根际土壤微生物对土壤改良发挥着重要作用。为了深入

探索环渤海滨海盐碱地碱蓬根际土壤细菌群落结构组成及其功能, 采用Illumina Misep高通量测序平台对环渤

海地区滨海盐碱地盐地碱蓬根际土壤和裸地土壤进行测序。从16个样本中获得有效序列734 792条, 4 285个

OTUs, 归属于41门、100纲、282目、400科、892属、1 577种。盐地碱蓬根际土壤细菌群落由变形菌门

(Proteobacteria)、放线菌门(Actinobacteria)、绿弯曲门(Chloroflexi)、拟杆菌门(Bacteroidetes)、芽单胞菌门

(Gemmatimonadetes)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicutes)、蓝藻细菌门(Cyanobacteria)、髌骨细菌

门(Patescibacteria、浮霉菌门(Planctomycetes)组成。Alpha多样性计算结果表明, 盐地碱蓬根际土壤细菌群落

结构多样性高并与裸地土壤间差异显著; LEfSe (LDA Effect Size)分析发现, 盐地碱蓬与裸地差异指示种明显

不同。PCoA与相关性Heatmap表明, 盐地碱蓬、速效氮、速效钾、速效磷、电导率是影响土壤细菌目类水平

群落组成的主要因子。PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved

States)分析表明微生物群落在新陈代谢等40个功能方面盐地碱蓬根际土壤比裸地土壤高。本研究表明盐地碱

蓬覆盖能够降低土壤盐分, 增加土壤养分, 对土壤细菌群落多样性及其功能有积极作用。

关键词: 高通量测序; 滨海盐碱地; 盐地碱蓬; 根际土壤; 细菌群落结构; 细菌群落功能

中图分类号: S154.36 开放科学码(资源服务)标识码(OSID):

The bacterial community structure and function of

Suaeda salsa rhizosphere soil

*

SUN Jianping, LIU Yahui

**

, ZUO Yongmei, HAN Minli, ZHANG Hongwei, LYU Jingjing

(Institute of Coastal Agricultural, Hebei Academy of Agriculture and Forestry Sciences / Saline and Alkali Land Greening

Engineering Technology Center of Heibei Province / Tangshan Key Laboratory of Plant Salt Tolerance Research, Tangshan 063299,

China)

Abstract: Suaeda salsa is an ideal agent for the biological enrichment of saline-alkali soil. Microorganisms in the rhizosphere

of this plant play an essential role in soil improvement. The Illumina Misep high-throughput sequencing platform was used to

explore the structural composition and function of the bacterial community in the rhizosphere soil of S. salsa and bare soil

from coastal saline-alkali land in the Bohai Bay Rim area of Hebei, Shandong, and Tianjin, China. In total, 734 792 effective

sequences were obtained from 16 samples, of which 4 285 OUTs belonged to 41 phyla, 100 classes, 282 orders, 400 families,

*

河北省农林科学院基本业务费(201810101)和河北省农林科学院现代农业科技创新工程项目(2019-1-6-2)资助

**

通信作者: 刘雅辉, 主要研究方向为盐碱地改良与农业高效利用。E-mail: bhslyh@

孙建平, 主要研究方向为滨海盐碱地改良与农业高效利用。E-mail: bhssjp@

收稿日期: 2020-03-04 接受日期: 2020-05-29

*

This study was supported by the Basic Business Expenses of Hebei Academy of Agricultural and Forestry Sciences (201810101), the Agri-

culture Science and Technology Innovation Project of Hebei Academy of Agricultural and Forestry Sciences (2019-1-6-2).

** Corresponding author, E-mail: bhslyh@

Received Mar. 4, 2020; accepted May 29, 2020

第

10

期

孙建平等: 盐地碱蓬根际土壤细菌群落结构及其功能 1619

892 genera, and 1 577 species. The bacterial community in the rhizosphere soil of S. salsa contained Proteobacteria, Actino-

bacteria, Chloroflexi, Bacteroidetes, Gemmatimonadetes, Acidobacteria, Firmicutes, Cyanobacteria, Patescibacteria, and

Planctomycetes. These results were consistent with the Alpha diversity analysis results, indicating that the community was

highly diversified and significantly different from that of the bare soil. The LEfSe (LDA Effect Size) analysis showed that

indicator species differentially occurred in S. salsa and bare soils. In S. salsa soil, Cyanobacteria, Acidobacteria, Alphapro-

teobacteria, Oxyphotobacteria, Chloroflexi, Rhizobiales, Nostocales, Sphingomonadales, Sphingomonadaceae, and Bacillus

were the indicator species. Based on principal coordinates analyses and a correlation heatmap, the main factors affecting the

soil bacterial community at order level were the presence of S. salsa, alkali-hydrolyzable nitrogen, available potassium, avail-

able phosphorus, and electrical conductivity. Also, Ectothiorhodospira and Balneolaceae could survive in bare soil with poor

fertility, high salinity, and a viscous structure. PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of

Unobserved States) analysis showed that 304 metabolic pathways at pathway level 3 were active in both soil, of which 41

pathways, especially those involving in metabolism were different between S. salsa soil and bare land soil. These results indi-

cated that S. salsa growth has a positive effect on the diversity and function of soil bacterial community by improving soil

structure and increasing nutrients levels. These findings may be applied to improve saline-alkali land, optimize soil environ-

ment, and enhance its usefulness and sustainability.

Keywords: High-throughput sequencing; Coastal saline-alkali land; Suaeda salsa; Rhizosphere soil; Bacterial community

structure; Bacterial community function

土壤微生物在生态系统功能中起重要的作用,

其多样性不仅能够描述任何一种复杂生态环境, 还

可以较早地反映该环境的变化, 已被认为是生态系

在土壤微生物中, 细菌的

统中重要的生物学指标

[1]

。

数量和种类最多、分布最广泛且代谢功能潜力巨大,

参与碳、氮物质循环、土壤结构形成、促进植物生

长以及改善生态环境等过程

[2]

。植物为微生物提供

营养物质, 如分泌物质、落叶、花粉脱落以及枯死

的植物残体, 微生物分解无机物为植物生长提供营

养, 这些过程均发生在根际, 因此研究根际土壤细

菌群落多样性及其功能, 有助于了解植物、土壤微

生物及其环境之间的关系。

滨海盐碱土壤属于极端环境, 其环境的复杂多

样性造就了物种丰富多样性, 物种的多样性预示着

物种功能的多样性。微生物独特的遗传特性、生理

功能影响着植物的生存环境

[3]

, 因此滨海盐碱土壤

是研究土壤微生物细菌群落与耐盐植物关系的天然

实验室。盐地碱蓬(Suaeda salsa)是藜科

(Chenopodiaceae)碱蓬属植物, 不但在食用和药用领

域备受瞩目, 也是用于开发利用盐碱地的优势植物

已有研究表明, 盐地碱蓬的根系能分泌有机

之一

[4]

。

酸, 植物残体分解后产生的腐殖酸能够降低土壤碱

性, 改善土壤理化性质, 减少土壤水分损失, 还能

增加土壤中微生物的数量和酶活性

[5-8]

。目前对于盐

地碱蓬的微生物研究多局限于纯培养法的测定与分

析, 而针对其微生物群落结构的研究报道很少, 尤

其是用分子生物学方法研究盐地碱蓬根际土壤微生

物群落结构及功能尚鲜有报道。因此明确盐地碱蓬

根际土壤中的细菌多样性优势物种及功能, 可为利

用盐地碱蓬根系原生耐盐微生物改善盐碱地微环境

质量, 实施安全有效的盐碱地生物改良措施提供土

壤微生物方面的理论依据。

近年来, 高通量测序技术发展迅速, 它能够快速

揭示原位环境中微生物群落的复杂性和多样性

[9-11]

。

本研究采用Misep高通量测序方法, 对盐地碱蓬根

际土壤细菌进行16s rRNA基因测序, 分析环渤海地

区滨海盐碱地盐地碱蓬根际土壤的细菌群落结构,

探索细菌群落结构与环境因子的关系, 结合

PICRUSt (Phylogenetic Investigation of Communities

by Reconstruction of Unobserved States)分析预测盐

地碱蓬根际土壤细菌的功能, 不仅为环渤海地区滨

海盐碱地盐地碱蓬的生长和生物改良措施的应用提

供支撑, 也为今后研发土壤改良剂奠定基础。

1 材料和方法

1.1 研究地点与样品采集

以河北、山东、天津的滨海盐碱地为研究区, 其

紧临渤海湾, 主要由海相沉积物和海陆交互层沉积

的黏性土、粉土及砂类土组成, 土壤类型为滨海盐

土。环渤海地区光照充足, 年日照时数2 400~3 000 h,

比山麓平原区多200 h左右; 年平均气温10.8~

13.3 ℃, 无霜期180~200 d; 低平原区年降水量

450~550 mm, 滨海平原区年降水量600~700 mm,

80%的降水主要集中在7—9月

[12]

。该区属暖温带半

湿润季风性气候, 光照充足, 四季分明, 其中春季

干旱少雨、夏季炎热多雨、秋季冷暖适中、冬季寒

2024年5月29日发(作者：长孙云天)

中国生态农业学报(中英文)

2020年10月 第28卷 第10期

Chinese Journal of Eco-Agriculture, Oct. 2020, 28(10): 16181629

DOI: 10.13930/.200160

孙建平, 刘雅辉, 左永梅, 韩民利, 张虹伟, 吕晶晶. 盐地碱蓬根际土壤细菌群落结构及其功能[J]. 中国生态农业学报

(中英文), 2020, 28(10):

16181629

SUN J P, LIU Y H, ZUO Y M, HAN M L, ZHANG H W, LYU J J. The bacterial community structure and function of Suaeda

salsa rhizosphere soil[J]. Chinese Journal of Eco-Agriculture, 2020, 28(10):

16181629

盐地碱蓬根际土壤细菌群落结构及其功能

孙建平, 刘雅辉

**

, 左永梅, 韩民利, 张虹伟, 吕晶晶

(河北省农林科学院滨海农业研究所/河北省盐碱地绿化工程技术中心/唐山市耐盐植物重点实验室 唐山 063299)

*

摘 要: 盐地碱蓬作为生物改良盐碱地的理想材料, 其根际土壤微生物对土壤改良发挥着重要作用。为了深入

探索环渤海滨海盐碱地碱蓬根际土壤细菌群落结构组成及其功能, 采用Illumina Misep高通量测序平台对环渤

海地区滨海盐碱地盐地碱蓬根际土壤和裸地土壤进行测序。从16个样本中获得有效序列734 792条, 4 285个

OTUs, 归属于41门、100纲、282目、400科、892属、1 577种。盐地碱蓬根际土壤细菌群落由变形菌门

(Proteobacteria)、放线菌门(Actinobacteria)、绿弯曲门(Chloroflexi)、拟杆菌门(Bacteroidetes)、芽单胞菌门

(Gemmatimonadetes)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicutes)、蓝藻细菌门(Cyanobacteria)、髌骨细菌

门(Patescibacteria、浮霉菌门(Planctomycetes)组成。Alpha多样性计算结果表明, 盐地碱蓬根际土壤细菌群落

结构多样性高并与裸地土壤间差异显著; LEfSe (LDA Effect Size)分析发现, 盐地碱蓬与裸地差异指示种明显

不同。PCoA与相关性Heatmap表明, 盐地碱蓬、速效氮、速效钾、速效磷、电导率是影响土壤细菌目类水平

群落组成的主要因子。PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of Unobserved

States)分析表明微生物群落在新陈代谢等40个功能方面盐地碱蓬根际土壤比裸地土壤高。本研究表明盐地碱

蓬覆盖能够降低土壤盐分, 增加土壤养分, 对土壤细菌群落多样性及其功能有积极作用。

关键词: 高通量测序; 滨海盐碱地; 盐地碱蓬; 根际土壤; 细菌群落结构; 细菌群落功能

中图分类号: S154.36 开放科学码(资源服务)标识码(OSID):

The bacterial community structure and function of

Suaeda salsa rhizosphere soil

*

SUN Jianping, LIU Yahui

**

, ZUO Yongmei, HAN Minli, ZHANG Hongwei, LYU Jingjing

(Institute of Coastal Agricultural, Hebei Academy of Agriculture and Forestry Sciences / Saline and Alkali Land Greening

Engineering Technology Center of Heibei Province / Tangshan Key Laboratory of Plant Salt Tolerance Research, Tangshan 063299,

China)

Abstract: Suaeda salsa is an ideal agent for the biological enrichment of saline-alkali soil. Microorganisms in the rhizosphere

of this plant play an essential role in soil improvement. The Illumina Misep high-throughput sequencing platform was used to

explore the structural composition and function of the bacterial community in the rhizosphere soil of S. salsa and bare soil

from coastal saline-alkali land in the Bohai Bay Rim area of Hebei, Shandong, and Tianjin, China. In total, 734 792 effective

sequences were obtained from 16 samples, of which 4 285 OUTs belonged to 41 phyla, 100 classes, 282 orders, 400 families,

*

河北省农林科学院基本业务费(201810101)和河北省农林科学院现代农业科技创新工程项目(2019-1-6-2)资助

**

通信作者: 刘雅辉, 主要研究方向为盐碱地改良与农业高效利用。E-mail: bhslyh@

孙建平, 主要研究方向为滨海盐碱地改良与农业高效利用。E-mail: bhssjp@

收稿日期: 2020-03-04 接受日期: 2020-05-29

*

This study was supported by the Basic Business Expenses of Hebei Academy of Agricultural and Forestry Sciences (201810101), the Agri-

culture Science and Technology Innovation Project of Hebei Academy of Agricultural and Forestry Sciences (2019-1-6-2).

** Corresponding author, E-mail: bhslyh@

Received Mar. 4, 2020; accepted May 29, 2020

第

10

期

孙建平等: 盐地碱蓬根际土壤细菌群落结构及其功能 1619

892 genera, and 1 577 species. The bacterial community in the rhizosphere soil of S. salsa contained Proteobacteria, Actino-

bacteria, Chloroflexi, Bacteroidetes, Gemmatimonadetes, Acidobacteria, Firmicutes, Cyanobacteria, Patescibacteria, and

Planctomycetes. These results were consistent with the Alpha diversity analysis results, indicating that the community was

highly diversified and significantly different from that of the bare soil. The LEfSe (LDA Effect Size) analysis showed that

indicator species differentially occurred in S. salsa and bare soils. In S. salsa soil, Cyanobacteria, Acidobacteria, Alphapro-

teobacteria, Oxyphotobacteria, Chloroflexi, Rhizobiales, Nostocales, Sphingomonadales, Sphingomonadaceae, and Bacillus

were the indicator species. Based on principal coordinates analyses and a correlation heatmap, the main factors affecting the

soil bacterial community at order level were the presence of S. salsa, alkali-hydrolyzable nitrogen, available potassium, avail-

able phosphorus, and electrical conductivity. Also, Ectothiorhodospira and Balneolaceae could survive in bare soil with poor

fertility, high salinity, and a viscous structure. PICRUSt (Phylogenetic Investigation of Communities by Reconstruction of

Unobserved States) analysis showed that 304 metabolic pathways at pathway level 3 were active in both soil, of which 41

pathways, especially those involving in metabolism were different between S. salsa soil and bare land soil. These results indi-

cated that S. salsa growth has a positive effect on the diversity and function of soil bacterial community by improving soil

structure and increasing nutrients levels. These findings may be applied to improve saline-alkali land, optimize soil environ-

ment, and enhance its usefulness and sustainability.

Keywords: High-throughput sequencing; Coastal saline-alkali land; Suaeda salsa; Rhizosphere soil; Bacterial community

structure; Bacterial community function

土壤微生物在生态系统功能中起重要的作用,

其多样性不仅能够描述任何一种复杂生态环境, 还

可以较早地反映该环境的变化, 已被认为是生态系

在土壤微生物中, 细菌的

统中重要的生物学指标

[1]

。

数量和种类最多、分布最广泛且代谢功能潜力巨大,

参与碳、氮物质循环、土壤结构形成、促进植物生

长以及改善生态环境等过程

[2]

。植物为微生物提供

营养物质, 如分泌物质、落叶、花粉脱落以及枯死

的植物残体, 微生物分解无机物为植物生长提供营

养, 这些过程均发生在根际, 因此研究根际土壤细

菌群落多样性及其功能, 有助于了解植物、土壤微

生物及其环境之间的关系。

滨海盐碱土壤属于极端环境, 其环境的复杂多

样性造就了物种丰富多样性, 物种的多样性预示着

物种功能的多样性。微生物独特的遗传特性、生理

功能影响着植物的生存环境

[3]

, 因此滨海盐碱土壤

是研究土壤微生物细菌群落与耐盐植物关系的天然

实验室。盐地碱蓬(Suaeda salsa)是藜科

(Chenopodiaceae)碱蓬属植物, 不但在食用和药用领

域备受瞩目, 也是用于开发利用盐碱地的优势植物

已有研究表明, 盐地碱蓬的根系能分泌有机

之一

[4]

。

酸, 植物残体分解后产生的腐殖酸能够降低土壤碱

性, 改善土壤理化性质, 减少土壤水分损失, 还能

增加土壤中微生物的数量和酶活性

[5-8]

。目前对于盐

地碱蓬的微生物研究多局限于纯培养法的测定与分

析, 而针对其微生物群落结构的研究报道很少, 尤

其是用分子生物学方法研究盐地碱蓬根际土壤微生

物群落结构及功能尚鲜有报道。因此明确盐地碱蓬

根际土壤中的细菌多样性优势物种及功能, 可为利

用盐地碱蓬根系原生耐盐微生物改善盐碱地微环境

质量, 实施安全有效的盐碱地生物改良措施提供土

壤微生物方面的理论依据。

近年来, 高通量测序技术发展迅速, 它能够快速

揭示原位环境中微生物群落的复杂性和多样性

[9-11]

。

本研究采用Misep高通量测序方法, 对盐地碱蓬根

际土壤细菌进行16s rRNA基因测序, 分析环渤海地

区滨海盐碱地盐地碱蓬根际土壤的细菌群落结构,

探索细菌群落结构与环境因子的关系, 结合

PICRUSt (Phylogenetic Investigation of Communities

by Reconstruction of Unobserved States)分析预测盐

地碱蓬根际土壤细菌的功能, 不仅为环渤海地区滨

海盐碱地盐地碱蓬的生长和生物改良措施的应用提

供支撑, 也为今后研发土壤改良剂奠定基础。

1 材料和方法

1.1 研究地点与样品采集

以河北、山东、天津的滨海盐碱地为研究区, 其

紧临渤海湾, 主要由海相沉积物和海陆交互层沉积

的黏性土、粉土及砂类土组成, 土壤类型为滨海盐

土。环渤海地区光照充足, 年日照时数2 400~3 000 h,

比山麓平原区多200 h左右; 年平均气温10.8~

13.3 ℃, 无霜期180~200 d; 低平原区年降水量

450~550 mm, 滨海平原区年降水量600~700 mm,

80%的降水主要集中在7—9月

[12]

。该区属暖温带半

湿润季风性气候, 光照充足, 四季分明, 其中春季

干旱少雨、夏季炎热多雨、秋季冷暖适中、冬季寒