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培菌白蚁菌圃和粪便微生物多样性分析

张硕1 林子佳1 肖元玺1 李净净1 李枝1 倪金凤1

(1.山东大学微生物技术研究院微生物技术国家重点实验室, 山东青岛 266237)

【摘要】【背景】培菌白蚁是属于白蚁科的一类与鸡枞菌属真菌共生的高等白蚁,其与体内肠道微生物和体外菌圃微生物形成三维共生体系。【目的】分析培菌白蚁菌圃和粪便的微生物多样性,并与肠道微生物进行比较。【方法】通过Illumina MiSeq高通量测序方法对培菌白蚁菌圃和粪便样品进行细菌16S rRNA基因和真菌ITS测序分析。【结果】高通量测序获得培菌白蚁菌圃和粪便样品细菌和真菌的有效序列和OTU数目。5个样品细菌OTU数目在90-199之间,而真菌OTU在10-58之间,细菌的种类多样性明显大于真菌。不论是细菌还是真菌,粪便样品的OTU数目多于菌圃样品。经物种分类分析,菌圃样品主要优势细菌是变形菌门(Proteobacteria),其相对含量超过82.4%;其次是拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes);粪便样品中优势细菌为拟杆菌门,其次是变形菌门,粪便优势菌属为别样杆菌属和营发酵单胞菌属,这与培菌白蚁肠道菌多样性组成一致。培菌白蚁菌圃和粪便样品共生真菌主要为担子菌门(Basidiomycota)和子囊菌门(Ascomycota)。菌圃优势真菌为鸡枞菌属(Termitomyces),相对含量在51.83%以上,菌圃中还鉴定到炭角菌属(1%,Xylaria)。【结论】为今后培菌白蚁-体内外微生物共生关系研究以及微生物的分离培养提供了依据和参考。

【关键词】 培菌白蚁;菌圃;粪便;细菌和真菌多样性;

【DOI】

【基金资助】 国家自然科学基金(31970119,31272370);

Symbiotic microorganism diversity in comb and feces of fungus-growing termite

ZHANG Shuo1 LIN Zi-Jia1 XIAO Yuan-Xi1 LI Jing-Jing1 LI Zhi1 NI Jin-Feng1

(1.State Key Laboratory of Microbial Technology, Institute of Microbial Technology, Shandong University, Qingdao, Shandong, China 266237)

【Abstract】[Background] Fungus-growing termites are a kind of higher termites belonging to the Macrotermitinae that culture monofungus (Termitomyces spp.) in their nests. Fungus-growing termites form a three-dimensional symbiosis system with intestinal microorganisms and microorganisms in the fungus comb. [Objective] To analyze the microbial diversity of fungus combs and termite feces and compare with intestinal microorganisms of fungus-growing termites. [Methods] The bacterial 16S rRNA gene amplicon and fungal ITS rRNA were analyzed with the Illumina MiSeq high-throughput sequencing method. [Results] The number of filtered sequences and OTU numbers of bacteria and fungi in samples of fungus comb and feces was obtained. The number of bacterial OTUs in the five samples was in the range of 90–199, while that of the fungal OTUs was in the range of 10–58. The diversity of bacteria was significantly greater than that of fungi. The OTUs in bacteria and fungi in fecal samples were more than those in fungus comb samples. According to taxa classification, the dominant phylum in the fungus comb samples was Proteobacteria, with the relative abundance of over 82.4%, followed by Bacteroidetes and Firmicutes. The dominant bacteriaal phylum in fecal samples was Bacteroidetes, followed by Proteobacteria. The dominant bacteria belong to Alistipes and Dysgonomonas, which was consistent with the diversity of intestinal bacteria in fungus-growing termites. The dominant fungi in termite comb and fecal samples were mainly Basidiomycota and Ascomycota. The dominant fungi genus in fungus comb was Termitomyces, with the relative abundance of more than 51.83%. Xylaria was also identified in the comb. [Conclusion] This study provides a basis and reference for the future research on the symbiosis of fungus-growing termites with in vivo and in vitro microorganisms and the isolation and culture of microorganisms.

【Keywords】 Fungus-growing termites; Fungus comb; Feces; Bacterial and fungal diversity;

【DOI】

【Funds】 National Natural Science Foundation of China (31970119, 31272370);

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    References

    [1] Inward D, Beccaloni G, Eggleton P. Death of an order: a comprehensive molecular phylogenetic study confirms that termites are eusocial cockroaches [J]. Biology Letters, 2007, 3 (3): 331–335

    [2] Eggleton P. An introduction to termites: biology, taxonomy and functional morphology [A]//Bignell DE, Roisin Y, Lo N. Biology of Termites: A Modern Synthesis. Dordrecht: Springer, 2010: 1–26

    [3] Ni JF, Tokuda G. Lignocellulose-degrading enzymes from termites and their symbiotic microbiota [J]. Biotechnology Advances, 2013, 31 (6): 838–850

    [4] Hongoh Y. Toward the functional analysis of uncultivable, symbiotic microorganisms in the termite gut [J]. Cellular and Molecular Life Sciences, 2011, 68 (8): 1311–1325

    [5] Brune A. Symbiotic digestion of lignocellulose in termite guts [J]. Nature Reviews Microbiology, 2014, 12 (3): 168–180

    [6] Sun XX, Wei JH, Li JJ, et al. Whole-genome analysis of the dominant bacterium Dysgonomonas macrotermitis in the hindgut of Macrotermes barneyi [J]. Acta Microbiologica Sinica, 2018, 58 (6): 995–1003 (in Chinese)

    [7] Aanen DK, de Fine Licht HH, Debets AJM, et al. High symbiont relatedness stabilizes mutualistic cooperation in fungus-growing termites [J]. Science, 2009, 326 (5956): 1103–1106

    [8] Hinze B, Crailsheim K, Leuthold RH. Polyethism in food processing and social organisation in the nest of Macrotermes bellicosus (Isoptera, Termitidae) [J]. Insectes Sociaux, 2002, 49: 31–37

    [9] da Costa RR, Hu HF, Li HJ, et al. Symbiotic plant biomass decomposition in fungus-growing termites [J]. Insects, 2019, 10 (4): 87

    [10] Shinzato N, Muramatsu M, Matsui T, et al. Phylogenetic analysis of the gut bacterial microflora of the fungus-growing termite Odontotermes formosanus [J]. Bioscience, Biotechnology, and Biochemistry, 2007, 71 (4): 906–915

    [11] Otani S, Mikaelyan A, Nobre T, et al. Identifying the core microbial community in the gut of fungus-growing termites [J]. Molecular Ecology, 2014, 23 (18): 4631–4644

    [12] Sun XX, Li JJ, Du J, et al. Cellulomonas macrotermitis sp. nov., a chitinolytic and cellulolytic bacterium isolated from the hindgut of a fungus-growing termite [J]. Antonie Van Leeuwenhoek, 2018, 111 (3): 471–478

    [13] Hu HF, da Costa RR, Pilgaard B, et al. Fungiculture in termites is associated with a mycolytic gut bacterial community [J]. m Sphere, 2019, 4 (3): e00165–19

    [14] Sun XX, Ning N, Tan HJ, et al. Research progress of diversity and function of symbiotic microorganisms in the gut of termites [J]. Chinese Journal of Applied and Environmental Biology, 2017, 23 (4): 764–770 (in Chinese)

    [15] Li HJ, Dietrich C, Zhu N, et al. Age polyethism drives community structure of the bacterial gut microbiota in the fungus-cultivating termite Odontotermes formosanus [J]. Environmental Microbiology, 2016, 18 (5): 1440–1451

    [16] Liang SY, Wang CP, Ahmad F, et al. Exploring the effect of plant substrates on bacterial community structure in termite fungus-combs [J]. PLo S One, 2020, 15 (5): e0232329

    [17] Poulsen M, Hu HF, Li C, et al. Complementary symbiont contributions to plant decomposition in a fungus-farming termite [J]. Proceedings of the National Academy of Sciences of the United States of America, 2014, 111 (40): 14500–14505

    [18] Sun XX, Li JJ, Ning N, et al. Isolation and identification of chitin-degrading bacteria from the hindgut of Macrotermes barneyi [J]. Microbiology China, 2017, 44 (7): 1649–1654 (in Chinese)

    [19] Li HJ, Yelle DJ, Li C, et al. Lignocellulose pretreatment in a fungus-cultivating termite [J]. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114 (18): 4709–4714

    [20] Yang YJ, Zhang N, Ji SQ, et al. Dysgonomonas macrotermitis sp. nov., isolated from the hindgut of a fungus-growing termite [J]. International Journal Systematic and Evolutionary Microbiology, 2014, 64 (Pt_9): 2956–2961

    [21] Otani S, Zhukova M, Koné NA, et al. Gut microbial compositions mirror caste-specific diets in a major lineage of social insects [J]. Environmental Microbiology Reports, 2019, 11 (2): 196–205

    [22] Otani S, Challinor VL, Kreuzenbeck NB, et al. Disease-free monoculture farming by fungus-growing termites [J]. Scientific Reports, 2019, 9 (1): 8819

    [23] Mikaelyan A, Dietrich C, Köhler T, et al. Diet is the primary determinant of bacterial community structure in the guts of higher termites [J]. Molecular Ecology, 2015, 24 (20): 5284–5295

    [24] Vesala R, Harjuntausta A, Hakkarainen A, et al. Termite mound architecture regulates nest temperature and correlates with species identities of symbiotic fungi [J]. Peer J, 2019, 6: e6237

    [25] Nagam V, Aluru R, Shoaib M, et al. Diversity of fungal isolates from fungus-growing termite Macrotermes barneyi and characterization of bioactive compound from Xylaria escharoidea [J]. Insect Science, 2020. DOI: 10.1111/1744-7917.12799

    [26] Visser AA, Nobre T, Currie CR, et al. Exploring the potential for actinobacteria as defensive symbionts in fungus-growing termites [J]. Microbial Ecology, 2012, 63 (4): 975–985

    [27] Mathew GM, Ju YM, Lai CY, et al. Microbial community analysis in the termite gut and fungus comb of Odontotermes formosanus: the implication of Bacillus as mutualists [J]. FEMS Microbiology Ecology, 2012, 79 (2): 504–517

    [28] Yin CP, Jin LP, Li S, et al. Diversity and antagonistic potential of Actinobacteria from the fungus-growing termite Odontotermes formosanus [J]. 3 Biotech, 2019, 9 (2): 45

This Article

ISSN:0253-2654

CN: 11-1996/Q

Vol 47, No. 10, Pages 3091-3101

October 2020

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Article Outline

Abstract

  • 1 Materials and methods
  • 2 Results and analysis
  • 3 Discussion and conclusions
  • References