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板栗果实内生真菌群落组成及分化特征
引用本文:闾星,窦桂铭,汪慧华,李如华,江泽平,李志朋,严东辉. 板栗果实内生真菌群落组成及分化特征[J]. 林业科学研究, 2022, 35(4): 9-22. DOI: 10.13275/j.cnki.lykxyj.2022.004.002
作者姓名:闾星  窦桂铭  汪慧华  李如华  江泽平  李志朋  严东辉
作者单位:1.中国林业科学研究院森林生态环境与自然保护研究所 国家林业和草原局森林保护学重点实验室,北京 100091;2.北京农业职业学院 食品与生物工程系,北京 100085;3.北京市怀柔区园林绿化局,北京 101400
摘    要:目的 探明果实内生真菌群落结构组成和多样性是了解果实健康微生物组功能和机制的基础。 方法 采用高通量测序技术和传统组织分离方法,对6个板栗品种的健康果实及其果肉(子叶)和果壳(含种皮)组织的内生真菌群落进行了多样性和组成分析。 结果 6个品种的果实及其2个组织共36个样本总共产生3 229 205条高质量序列,注释出真菌1 557个OTUs,分属10个门,38个纲,97个目,203个科,350个属。板栗果实中子囊菌门Ascomycota(平均相对丰度80.40%)丰度最高,其次为担子菌门Basidiomycota(平均相对丰度7.70%),优势纲为粪壳菌纲Sordariomycetes(平均相对丰度43.00%)和酵母纲Saccharomycetes(平均相对丰度15.10%)。品种间共有(Shared)真菌类群富集在座囊菌纲Dothideomycetes上,其丰度>90%。板栗果实品种间,子囊菌门在每一个品种中同样体现为最丰富类群,相对丰度为57.96%~90.15%,其次为担子菌门,相对丰度为3.28%~23.71%;纲水平上,优势类群出现变化,酵母菌纲真菌相对丰度(39.21%)在毛板红品种中是最高的,而粪壳菌纲真菌是另外5个板栗品种中最丰富类群(29.47%~59.56%)。果实组织间,果肉组织较之果壳具有更高的特有真菌多样性,果肉组织特有优势类群全部为担子菌,尤其是银耳纲Tremellomycetes的红菇目Russulales。LEfSe分析显示,果壳和果肉组织存在显著性差异群落组成或种群,镰孢属Fusarium、梅奇酵母属Metschnikowia、迈耶氏酵母属Meyerozyma和德巴利酵母属Debaryomyces真菌在果肉组织中具有显著优势。采用人工培养方法共获得有效菌株2 346株,分属子囊菌、担子菌和毛霉菌门Mucoromycota。除酵母菌类需特殊培养基筛选外,可培养方法显示果实及其果壳、果肉上的真菌优势类群,在群落组成和丰度上与高通量测序技术结果相似。 结论 板栗果实具有丰富的以子囊菌粪壳菌纲和酵母菌纲为优势类群的内生真菌群落组成和种群多样性,但建群种类主要来自座囊菌纲。板栗果实品种间的内生真菌群落组成结构具有明显差异。板栗果肉较之果壳组织有更为丰富的特有内生真菌多样性。板栗果肉和果壳组织内生真菌群落组成具有明显分化。本文系统报道了板栗果实内生真菌组的组成结构,为利用板栗健康微生物组防控果实病害和了解坚果类果实微生物组提供了基础。

关 键 词:板栗果实   内生真菌组   多样性   子叶   果皮   非培养技术
收稿时间:2022-02-14

Community Composition and Differentiation of Endophytic Mycobiota in Chestnut Fruits
LV Xing,DOU Gui-ming,WANG Hui-hua,LI Ru-hua,JIANG Ze-ping,LI Zhi-peng,YAN Dong-hui. Community Composition and Differentiation of Endophytic Mycobiota in Chestnut Fruits[J]. Forest Research, 2022, 35(4): 9-22. DOI: 10.13275/j.cnki.lykxyj.2022.004.002
Authors:LV Xing  DOU Gui-ming  WANG Hui-hua  LI Ru-hua  JIANG Ze-ping  LI Zhi-peng  YAN Dong-hui
Affiliation:1. Key Laboratory of Forest Protection of National Forestry and Grassland Administration, Ecology and Nature Conservation Institute, Chinese Academy of Forestry, Beijing 100091, China;2. Department of Food and Biological Engineering, Beijing Vaocational College of Agriculture, Beijing 100085, China;3. Beijing Huairou District Landscaping Bureau, Beijing 101400, China
Abstract:Objective Exploring the composition and diversity of the fruit endophytic fungi community is the basis for understanding the functions and mechanisms of fruit healthy microbiome. Method The diversity and composition of endophytic fungal communities in healthy fruit and its kernel (cotyledon) and shell (including seedcoat) parts of six chestnut cultivars were analyzed using high-throughput sequencing technology and traditional tissue isolation methods. Result A total of 3,229,205 high-quality sequences were generated from 36 samples of fruits and 2 tissues of six cultivars, and 1,557 fungal OTUs were annotated, belonging to 10 phyla, 38 classes, 97 orders, 203 families, and 350 genera. In chestnut fruit, Ascomycota (average relative abundance 80.40%) had the highest abundance, followed by Basidiomycota (average relative abundance 7.70%), and the dominant class were Sordariomycetes (average relative abundance 43.00%) and Saccharomycetes (average relative abundance 15.10%). The shared fungal taxa among species was enriched in Dothideomycetes with abundance >90%. Among chestnut fruit varieties, Ascomycota was also the most abundant group in each variety, with a relative abundance of 57.96%~90.15%, followed by Basidiomycota(3.28%~23.71%); At the class level, the dominant groups changed. The relative abundance of Saccharomycetes fungi (39.21%) was the highest in Maobanhong varieties, while Sordariomycetes fungi were the most abundant group (29.47%~59.56%) among the other five chestnut cultivars. Among the fruit tissues, the kernel tissue had higher unique fungal diversities than the shell, and the unique dominant groups of kernel tissue were all Basidiomycetes, especially the Russulales of Tremellomycetes. LEfSe analysis showed that there were significant differences in community composition or populations in the shell and kernel tissue, with Fusarium, Metschnikowia, Meyerozyma, and Debaryomyces fungi having significant dominance in kernel tissue. A total of 2346 effective strains were obtained by artificial culturable method, which belonged to Ascomycetes, Basidiomycetes and Mucoromycota. Except for yeasts that need special medium to be screened, the dominant fungal groups on fruits and tissues obtained by artificial culturing methods were similar to the results of high-throughput sequencing technology in terms of community composition and abundance. Conclusion Chestnut fruit has abundant endophytic fungal community composition and population diversity dominated by Sordariomycetes and Saccharomycetes, but the community-building species are mainly from Dothieomycetes. The endophytic fungal community composition is significantly different among chestnut fruit cultivars. Chestnut kernel has a richer diversity of exclusive endophytic fungi than shell part. There is a obvious differentiation on the composition of endophytic fungal community between chestnut kernel and shell tissue. This paper is the first systematic report on the compositional structure of endophytic fungi microbiota in chestnut fruit, which provides a basis for using chestnut healthy microbiome to prevent and control fruit diseases and to understand the nut fruit microbiome.
Keywords:chestnut fruits  endophytic fungal mycobiota  diversity  kernel  shell  culture-independent technique
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