白及根腐病植株根际土壤微生物群落组成特征分析

doi:10.13560/j.cnki.biotech.bull.1985.2021-0757

摘要/Abstract

摘要：

分析白及根腐病植株根际土壤微生物群落组成特征,为理解白及根腐病发病机理并针对性开展防治工作提供科学依据。利用Illumina HiSeq高通量测序技术,比较分析白及根腐病与健康植株根际土壤微生物群落结构特征,并对土壤理化性质与酶活性进行定量分析。白及根际土壤中优势真菌为子囊菌门（Ascomycota）和Mortierellomycota,优势细菌为变形菌门（Proteobacteria）和拟杆菌门（Bacteroidetes）。与健康植株相比,发病白及根际土壤Ascomycota相对丰度显著升高,而Mortierellomycota相对丰度显著降低;镰刀菌属（Fusarium）和白粉菌属（Erysiphe）等致病菌群相对丰度显著升高（P<0.05）。多元统计分析表明,健康与发病白及根际土壤细菌与真菌群落存在明显分化。此外,健康植株根际土壤脲酶、蛋白酶与蔗糖酶活性和有效钾含量较高,pH、有机质、铵态氮与硝态氮含量较低。白及根腐病的发生可能与土壤致病菌相对丰度升高、微生物群落结构变化、酶活性降低、养分失衡等因素有关。

关键词: 白及, 根腐病, 微生物群落, 高通量测序

Abstract:

Analyzing the composition features of microbial community in the rhizospheric soil of Bletilla striata with root rot would provide the scientific basis for understanding the microbial mechanisms leading to the root rot of B. striata and taking corresponding prevention and control measures. The Illumina HiSeq high-throughput sequencing was used to compare and analyze the microbial community structure in the rhizospheric soils of root-rot and healthy B. striata. Also,the soil physicochemical properties and enzyme activities were quantitatively analyzed. The dominant fungi were Ascomycota and Mortierellomycota,and the dominant bacteria were Proteobacteria and Bacteroidetes in the rhizospheric soil of B. striata. Compared with the healthy individuals,the relative abundance of Ascomycota significantly increased,while the relative abundance of Mortierellomycota decreased. The relative abundance of pathogenic Fusarium and Erysiphe significantly increased in the rhizospheric soil of root-rot B. striata（P<0.05）. Multivariate statistical analyses showed the significant differentiation of bacterial and fungal community in the rhizospheric soils between healthy and root-rot B. striata. In addition,the activities of urease,protease and sucrase,and available potassium content were higher in the rhizospheric soil of healthy individuals;while soil pH and the contents of organic matter,NH₄⁺-N,and NO₃^--N were lower. The root-rot disease of B. striata could be related to the increased relative abundance of pathogenic microorganisms,the changes of microbial community structure,the decreased soil enzyme activity,and the imbalance of plant nutrients.

Key words: Bletilla striata, root-rot, soil microbial community, high-throughput sequencing

赵林艳, 官会林, 向萍, 李泽诚, 柏雨龙, 宋洪川, 孙世中, 徐武美. 白及根腐病植株根际土壤微生物群落组成特征分析[J]. 生物技术通报, 2022, 38(2): 67-74.

ZHAO Lin-yan, GUAN Hui-lin, XIANG Ping, LI Ze-cheng, BAI Yu-long, SONG Hong-chuan, SUN Shi-zhong, XU Wu-mei. Composition Features of Microbial Community in the Rhizospheric Soil of Bletilla striata with Root Rot[J]. Biotechnology Bulletin, 2022, 38(2): 67-74.

图/表 6

表1 白及根腐病和健康植株根际土壤微生物门水平的相对丰度

Table 1 Relative abundance of microorganisms at the phylum level in the rhizospheric soils of root-rot and healthy B. striata

真菌Fungi			细菌Bacteria
Phylum	D	H	Phylum	D	H
Ascomycota	73.43%±6.59%^a	40.92%±10.96%^b	Proteobacteria	24.71%±1.62%^a	28.36%±1.76%^a
Mortierellomycota	11.78%±4.19%^b	42.50%±9.86%^a	Bacteroidetes	28.24%±2.35%^a	22.57%±2.90%^a
Basidiomycota	6.90%±0.90^a	12.08±7.47%^a	Acidobacteria	12.78%±1.458%^a	15.85%±2.525%^a
Chytridiomycota	0.39%±0.20%^a	0.12%±0.078%^a	Firmicutes	13.68%±1.62%^a	9.05%±1.82%^a
Rozellomycota	0.10%±0.042%^a	0.028%±0.023%^a	Actinobacteria	7.71%±0.86%^a	8.29%±0.98%^a
Olpidiomycota	0.083%±0.054%^a	0.012%±0.009%^a	Gemmatimonadetes	5.25%±0.49%^a	6.42%±0.61%^a
Others	7.32%±1.95%^a	4.34%±0.69%^a	Others	7.64%±0.29%^a	9.47%±0.79%^a

图1 白及根腐病和健康植株根际土壤微生物属分类水平的相对丰度仅展示具有显著差异的微生物类群（P<0.05）。图a-f属真菌类群,g-i属细菌类群。D表示根腐病组,H表示健康组

Fig. 1 Relative abundance of rhizospheric microorganisms at the genus level in the rhizospheric soils of root-rot and healthy B. striata Only the microbial taxa with significant difference are displayed（P<0.05）;Figs. a-f belong to the fungi and g-i belong to bacteria. D indicates the root-rot disease group,and H indicates the healthy group

图2 白及根腐病和健康植株根际土壤共有和特有的OTU数量 a代表真菌群落,b代表细菌群落

Fig. 2 Numbers of shared and unique OTUs in the rhizospheric soils of root-rot and healthy B. striata a indicates fungal community,and b indicates bacterial community

表2 白及根腐病和健康植株根际土壤微生物群落alpha多样性指数

Table 2 Alpha diversity of microbial community in the rhizospheric soils of root-rot and healthy B. striata

分组 Group	真菌Fungi			细菌Bacteria
分组 Group	OTU richness	Chao1	Shannon	OTU richness	Chao1	Shannon
D	639.8±40.30^a	675.09±45.02^a	3.82±0.58^a	1 607.20±11.03^a	1 647.26±11.19^a	6.45±0.02^a
H	526.0±6.31^b	639.43±38.54^a	2.92±0.33^a	1 619.00±13.17^a	1 668.55±8.14^a	6.37±0.04^a

图3 基于聚类与冗余分析的白及根腐病和健康植株根际土壤微生物群落差异特征图a和b为真菌群落聚类与冗余分析结果,c和d为细菌群落聚类与冗余分析结果

Fig. 3 Differences in the rhizospheric soil microbial community of root-rot and healthy B. striata based on clustering and redundancy analyses313 a and b show the results of clustering and redundancy analyses of fungal community,c and d show the results of clustering and redundancy analyses of bacterial community

表3 白及根腐病和健康植株根际土壤理化性质与酶活性

Table 3 Physicochemical properties and enzyme activities in the rhizospheric soils of root-rot and healthy B. striata

土壤因子Soil factor	H	D	P
pH	6.57±0.19	6.87±0.18	<0.05
EC（μS·cm^-1）	82.06±5.25	65.80±4.14	<0.05
铵态氮（mg·kg^-1）	4.35±0.42	5.12±0.36	<0.05
硝态氮（mg·kg^-1）	3.02±0.35	6.49±0.60	<0.01
有机质（g·kg^-1）	4.84±0.44	5.68±0.52	<0.05
有效磷（mg·kg^-1）	81.36±17.01	77.05±28.64	0.78
有效钾（mg·kg^-1）	414.12±14.71	358.00±15.81	<0.05
脲酶（mg·NH₄⁺-N·g^-1 24 h^-1）	0.51±0.03	0.46±0.03	<0.05
磷酸酶（mg phenol·g^-1·12 h^-1）	0.14±0.01	0.13±0.01	0.13
蛋白酶（mg·NH₂-N·g^-1·24 h^-1）	0.044±0.01	0.036±0.01	<0.01
蔗糖酶（mg glucose·g^-1·24 h^-1）	22.49±0.89	15.90±0.80	<0.01

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