马铃薯健株与黄萎病株根际土壤真菌群落结构及其对碳源利用特征

doi:10.3864/j.issn.0578-1752.2021.02.006

中国农业科学 ›› 2021, Vol. 54 ›› Issue (2): 296-309.doi: 10.3864/j.issn.0578-1752.2021.02.006

马铃薯健株与黄萎病株根际土壤真菌群落结构及其对碳源利用特征

赵卫松(),郭庆港,苏振贺,王培培,董丽红,胡卿,鹿秀云,张晓云,李社增,马平()

河北省农林科学院植物保护研究所/河北省农业有害生物综合防治工程技术研究中心/农业农村部华北北部作物有害生物综合治理重点实验室,河北保定 071000

收稿日期:2020-04-16 接受日期:2020-05-19 出版日期:2021-01-16 发布日期:2021-02-03
通讯作者: 马平
作者简介:赵卫松,Tel：0312-5927076;E-mail： zhaoweisong1985@163.com。
基金资助:
国家重点研发计划(2017YFD0200601);国家公益性行业(农业)科研专项(201503109);河北省农林科学院现代农业科技创新工程(1-02-03)

Characterization of Fungal Community Structure in the Rhizosphere Soil of Healthy and Diseased-Verticillium Wilt Potato Plants and Carbon Source Utilization

ZHAO WeiSong(),GUO QingGang,SU ZhenHe,WANG PeiPei,DONG LiHong,HU Qing,LU XiuYun,ZHANG XiaoYun,LI SheZeng,MA Ping()

Plant Protection Institute of Hebei Academy of Agricultural and Forestry Sciences/IPM Centre of Hebei Province/Key Laboratory of IPM on Crops in Northern Region of North China, Ministry of Agriculture and Rural Affairs, Baoding 071000, Hebei

Received:2020-04-16 Accepted:2020-05-19 Online:2021-01-16 Published:2021-02-03
Contact: Ping MA

摘要/Abstract

摘要：

【目的】通过研究马铃薯健康植株与黄萎病株的根际土壤真菌群落结构与功能多样性的差异,明确土壤真菌群落结构与黄萎病发生之间的关系,为最终从微生物生态学的角度解释马铃薯黄萎病的发生原因及其生态防控提供理论依据。【方法】以河北省坝上地区马铃薯健株与黄萎病株的根际土壤为研究对象,分别利用实时荧光定量PCR（real-time PCR）和高通量测序（Illumina MiSeq）技术检测根际土壤中大丽轮枝菌（Verticillium dahliae）ITS基因拷贝数量并分析真菌群落结构变化,结合冗余分析（RDA）明确真菌群落结构与土壤养分的相关性。同时利用Biolog-ECO平板法比较健株与黄萎病株根际土壤微生物对碳源的利用能力。【结果】马铃薯黄萎病的发生与土壤中大丽轮枝菌ITS基因拷贝数量存在相关性,在病株根际土壤中病原菌数量高,而在健株根际土壤中未检测到病原菌。高通量测序分析表明,病株根际土壤真菌多样性指数低于健康植株,但多样性差异不显著。在群落组成的门水平上,与健株根际土壤相比,病株根际土壤中的子囊菌门（Ascomycota）和丝孢菌门（Mortierellomycota）相对丰度上升幅度分别为20.68%和16.16%,而担子菌门（Basidiomycota）的相对丰度下降51.43%。在属水平上,病株根际土壤中轮枝菌属（Verticillium）、青霉属（Penicillium）、维希尼克氏酵母属（Vishniacozyma）、红酵母属（Rhodotorula）和芽枝霉属（Cladosporium）的相对丰度呈上升趋势,增加倍数分别为71.96、3.62、6.11、15.38和6.24倍,而小不整球壳属（Plectosphaerella）、Guehomyces、葡萄穗霉属（Stachybotrys）、赤霉属（Gibberella）、曲霉属（Aspergillus）菌群的相对丰度下降幅度分别为45.10%、61.41%、96.87%、45.85%和44.39%。真菌群落组成与土壤养分的冗余分析（RDA）表明,健株根际土壤优势群落的相对丰度（如小不整球壳属、Guehomyces、葡萄穗霉属、赤霉属、曲霉属）与硝态氮、有机质和pH呈正相关,而黄萎病株根际土壤优势群落的相对丰度（如轮枝菌属、链格孢属、刺盘孢属、被孢霉属、腐质霉属、青霉属、维希尼克氏酵母属、红酵母属和芽枝霉属）与无机磷和速效磷呈正相关。不同根际土壤的AWCD值表明,病株根际土壤微生物对碳源的利用能力高于健株。进一步分析发现,与健株相比,病株土壤微生物对羧酸类碳源的利用能力显著提高,而对氨基酸类、胺类、碳水化合物类、聚合物类和双亲化合物类碳源利用能力差异不显著。【结论】病株的根际土壤真菌多样性降低和群落结构改变是马铃薯黄萎病发生的重要特征,其中轮枝菌属菌群的相对丰度显著提高是最主要特征,并且真菌群落结构受土壤养分影响。同时,病株根际土壤微生物对羧酸类碳源利用能力显著提高。

关键词: 马铃薯黄萎病, 大丽轮枝菌, 根际土壤, 真菌群落结构, 高通量测序, 土壤养分, Biolog技术

Abstract:

【Objective】The objective of this study is to research the differences of the structure and functional diversity of fungal community in the rhizosphere soil of healthy and diseased potato plants, and to clarify the relationship between the structure of soil fungal community and the occurrence of verticillium wilt, so as to provide a theoretical basis for the final explanation of the causes of potato verticillium wilt and ecological control from the perspective of microbial ecology.【Method】The rhizosphere soil of healthy and diseased plants of verticillium wilt of potato was collected from Bashang area in Hebei Province. The number of ITS gene copies of Verticillium dahliae was determined by real-time quantitative PCR, and the structure of soil fungal communities was measured by high-throughput sequencing (Illumina MiSeq). The quantity of V. dahliae and the community of soil fungi were analyzed. The redundancy analysis (RDA) was used to determine the correlation between fungal community structure and soil nutrient. Meanwhile, the utilization capacity of rhizosphere soil microorganisms of healthy and diseased plants to carbon source was compared by Biolog-Eco plate method.【Result】The occurrence of verticillium wilt of potato was related to the ITS gene copy number of V. dahliae in soil. The ITS gene copy number of pathogens was high in rhizosphere soil of diseased plants, but not detected in healthy plants. Illumina MiSeq analysis showed that the diversity index of fungi in rhizosphere soil of diseased plants was lower than that of healthy plants, but the diversity difference was not significant. Compared with the rhizosphere soil of healthy plants, at phylum level, the relative abundance of Ascomycota and Mortierellomycota was increased by 20.68% and 16.16%, respectively, while that of Basidiomycota was decreased by 51.43%. At genus level, the average relative abundance of Verticillium, Penicillium, Vishniacozyma, Rhodotorula and Cladosporium in rhizosphere soil of diseased plants was increased by 71.96, 3.62, 6.11, 15.38 and 6.24 times, respectively, while that of Plectosaphaerella, Guehomyces, Stachybotrys, Gibberella, Aspergillus was decreased by 45.10%, 61.41%, 96.87%, 45.85% and 44.39%, respectively. RDA of fungal community composition and soil nutrient showed that the relative abundance of dominant communities in rhizosphere soil of healthy plants (such as Plectosaphaerella, Guehomyces, Grapevine, Gibberella and Aspergillus) was positively correlated with nitrate nitrogen (NO₃ ^--N), organic matter (OM) and pH, while the relative abundance of dominant communities in rhizosphere soil of diseased plants (such as Verticillium, Alternaria, Colletotrichum, Mortierella, Humicola, Penicillium, Vishniacozyma, Rhodotorula and Cladosporium) was positively correlated with inorganic phosphorus (IP) and available phosphorus (AP). The AWCD values of rhizosphere soil of healthy and diseased plants indicated that the utilization ability of microorganisms to carbon source in rhizosphere soil of diseased plants was higher than that of healthy plants. Further analysis showed that, compared with the healthy plants, the utilization ability of rhizosphere soil microorganisms to carboxylic acids carbon source was significantly improved, while the utilization ability to amino acids, amines, carbohydrates, polymers and miscellaneous was not significantly different.【Conclusion】The decrease of fungal diversity and the change of community structure in rhizosphere soil of healthy and diseased plants are the important characteristics of potato verticillium wilt. The significant increase of relative abundance of Verticillium is the most important characteristic, and the structure of fungal community is affected by soil nutrient. At the same time, the utilization of carboxylic acids carbon source is significantly increased by rhizosphere soil microorganisms of diseased plants.

Key words: potato verticillium wilt, Verticillium dahliae, rhizosphere soil, fungal community structure, Illumina MiSeq, soil nutrient, Biolog technique

赵卫松,郭庆港,苏振贺,王培培,董丽红,胡卿,鹿秀云,张晓云,李社增,马平. 马铃薯健株与黄萎病株根际土壤真菌群落结构及其对碳源利用特征[J]. 中国农业科学, 2021, 54(2): 296-309.

ZHAO WeiSong,GUO QingGang,SU ZhenHe,WANG PeiPei,DONG LiHong,HU Qing,LU XiuYun,ZHANG XiaoYun,LI SheZeng,MA Ping. Characterization of Fungal Community Structure in the Rhizosphere Soil of Healthy and Diseased-Verticillium Wilt Potato Plants and Carbon Source Utilization[J]. Scientia Agricultura Sinica, 2021, 54(2): 296-309.

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指标 Index	健株Healthy plant	病株Diseased plant
硝态氮NO₃^--N (mg·kg^-1)	13.38±4.48a	10.03±7.69b
pH	6.89±0.02a	6.83±0.03a
速效磷AP (mg·kg^-1)	373.51±60.03b	897.00±20.08a
无机磷IP (μg·g^-1)	99.40±11.71b	189.84±18.26a
有机质OM (%)	0.77±0.01a	0.64±0.01b
数据后不同小写字母表示差异显著（P<0.05）。下同 Different lowercases after the data indicate significant difference (P<0.05). The same as below

处理 Treatment	Sobs 指数 Sobs index	香浓指数 Shannon index	辛普森指数 Simpson index	ACE 指数 ACE index	Chao I 指数 Chao I index
健株Healthy plant	236.50±21.42a	2.5066±0.2077a	0.1753±0.0364a	291.44±17.11a	292.95±20.21a
病株Diseased plant	222.00±15.10a	2.4829±0.2238a	0.1730±0.0504a	280.17±25.83a	278.20±27.49a

马铃薯健株与黄萎病株根际土壤真菌群落结构及其对碳源利用特征

Characterization of Fungal Community Structure in the Rhizosphere Soil of Healthy and Diseased-Verticillium Wilt Potato Plants and Carbon Source Utilization

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