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嗜碱假单胞菌Ej2浸种对盐胁迫下水稻根系内生菌群落结构的影响
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宁夏回族自治区自然科学基金(2020AAC03317);宁夏回族自治区青年拔尖人才培养工程


Effects of soaking seeds with Pseudomonas alcaliphila Ej2 on endophytic community of rice roots under salt stress
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    摘要:

    【目的】为明确假单胞菌(Pseudomonas)浸种对盐胁迫下水稻根系内生菌群落多样性及结构的影响。【方法】提取宁粳61号根系内生细菌和内生真菌的总DNA后,利用高通量测序技术检测了水稻幼苗根系内生细菌的V5–V7高变区16S rRNA基因和内生真菌的ITS1–ITS2区ITS基因序列,分析了假单胞菌浸种处理对盐胁迫下水稻根系内生菌的微生态调控作用。【结果】结果表明,嗜碱假单胞菌(P. alcaliphila) Ej2浸种处理可以显著提高盐胁迫下水稻幼苗根系内生细菌和内生真菌群落的多样性、丰富度和均匀度,增加了内生菌门、纲、目、科、属和分类单元OTU (operational taxonomic units)(序列数≥5)的数量;增加内生细菌中拟杆菌门(Bacteroidetes)和放线菌门(Actinobacteria)的丰度,增加率分别达45.97%和9.55%,降低了厚壁菌门(Firmicutes)丰度,降低率为12.22%;显著降低内生真菌中子囊菌门(Ascomycota)的丰度,降低率为17.96%,显著增加被孢霉门(Mortierellomycota)和油壶菌门(Olpidiomycota)的丰度,油壶菌门是假单胞菌浸种处理后的独有真菌门。嗜碱假单胞菌Ej2浸种增加了盐胁迫下水稻幼苗根系内生细菌中假单胞菌属、根瘤菌属(Allorhizobium)、黄杆菌属(Flavobacterium)和德沃斯氏菌属(Devosia)的丰度;降低了内生真菌中光黑壳属(Preussia)、裂壳属(Schizothecium)、镰孢菌(Fusarium)、曲霉属(Aspergillus)和Monosporascus的丰度,油壶菌属(Olpidium)是假单胞菌浸种处理后的独有真菌属;对内生细菌种水平的影响表现为增加了施氏假单胞菌(P. stutzeri)、嗜碱假单胞菌、未分类假单胞菌及根瘤菌(Rhizobium)、不能培养根瘤菌的丰度,降低了黄色氢噬菌(Hydrogenophaga flava)的丰度;对内生真菌种水平的影响包括降低了栖土光黑壳(P. terricola)、Neocosmospora rubicola、Gibberella intricans、帚状曲霉(A. penicillioides)及未分类镰刀菌的丰度。PICRUSt2功能预测分析表明,嗜碱假单胞菌Ej2浸种增加了盐胁迫下水稻幼苗根系参与各种代谢通路的内生细菌丰度;FUNGuild功能预测分析显示,增加了动物病原真菌代谢相关的真菌丰度、菌根真菌丰度和内生真菌的丰度,降低了粪便浮生菌、真菌浮生物和植物病原菌的丰度。【结论】假单胞菌浸种可以显著提高盐胁迫下水稻幼苗根系内生菌群落多样性、改变优势菌群结构和改善盐胁迫下水稻幼苗根系生长的微生态环境。

    Abstract:

    [Objective] To reveal the impact of soaking seeds with fermentation broth of Pseudomonas alcaliphila Ej2 on the diversity and structure of endophytic community in rice roots under salt stress. [Methods] The total DNA of the endophytic bacteria and fungi of the roots of Ningjing 61 was extracted, followed by high-throughput sequencing of the V5–V7 regions of the bacterial 16S rDNA gene and the ITS1–ITS2 of fungal ITS region. Thereby, the regulatory effects of soaking seeds with Ej2 on endophytic community of rice roots under salt stress were analyzed.[Results] Soaking seeds with Ej2 significantly improved the diversity, richness, and evenness of endophytic community of rice roots under salt stress. To be specific, it increased the number of phyla, classes, families, genera, and operational taxonomic units (OTUs, read number≥5) of the endophytic community. In addition, for bacteria, it raised the relative abundance of Bacteroidetes (by 45.97%) and Actinobacteria (by 9.55%), and lowered the relative abundance of Firmicutes (by 12.22%). As to fungi, it significantly decreased relative abundance of Ascomycota (by 17.96%) and significantly elevated the relative abundance of Mortierellomycota andOlpidiomycota. Among them, Olpidiomycota was only detected after seed soaking with Ej2. After seeding soaking with Ej2, the relative abundance of Pseudomonas,Allorhizobium, Flavobacterium, and Devosia rose, and that of Preussia, Schizothecium, Fusarium, Aspergillus, and Monosporascus decreased. Olpidium was identified only after seed soaking with Ej2. As for the bacterial species, the relative abundance of P.stutzeri, P.alcaliphila, unclassified Pseudomonas, unclassified Rhizobium, and uncultured Rhizobium was up-regulated, and that of Hydrogenophaga flava was decreased after seeding soaking with Ej2. In terms of the fungal species the relative abundance of P.terricola, Neocosmospora rubicola, Gibberella intricans, A.penicillioides and unclassified Fusarium was reduced after seeding soaking with Ej2. PICRUSt2 prediction suggested that seeding soaking with Ej2 increased the abundance of bacterial community involved in various metabolic pathways in rice roots under salt stress, and FUNGuild analysis indicated the increase in the abundance of fungi related to the metabolism of animal pathogenic fungi, mycorrhizal fungi, and endophytic fungi, and reduction in relative abundance of saprophytic fecal community, fungal saprophytes, and plant pathogens.[Conclusion]Soaking seeds with Pseudomonas can significantly increase the diversity of endophytic community of rice roots under salt stress, change the structure of dominant endophytic flora, and improve the microecological environment of rice roots.

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沙月霞. 嗜碱假单胞菌Ej2浸种对盐胁迫下水稻根系内生菌群落结构的影响. 微生物学报, 2022, 62(5): 1919-1935

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  • 收稿日期:2021-10-29
  • 最后修改日期:2021-12-07
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  • 在线发布日期: 2022-04-30
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