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重度盐碱地细菌的资源分离培养方法
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中国农业科学院科技创新工程(CAAS-ZDRW202407);中央民族大学本科生研究训练计划(URTP2022110078)


Methods for isolating bacteria from heavy saline-alkali land
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    摘要:

    【背景】盐碱地是后备耕地资源,对农业粮食安全至关重要,施用植物促生菌是治理盐碱地的有效策略之一。【目的】探究能高效分离盐碱地细菌资源的方法,促进盐碱地植物促生菌的开发利用。【方法】以山东省东营市重度盐碱地大豆根际土为材料,采用平板稀释培养法分离细菌,考察有机营养水平、pH和NaCl浓度对盐碱地细菌分离培养的影响;依托功能平板筛选具有溶解无机磷和产铁载体能力的细菌;通过16S rRNA基因测序对纯化的菌株进行鉴定。【结果】共有443个分离株的16S rRNA基因测序成功,包含48个属96个种,潜在新种7个。筛选获得28种具有溶解无机磷功能的细菌,13种产生铁载体功能的细菌,同时有7种具备2种功能的细菌。从细菌种类的丰富度和CFU数量角度来看,影响强度顺序为NaCl浓度>有机营养水平>pH。在寡有机营养条件下,NaCl浓度≥3%时对盐碱地细菌的生长有明显的抑制作用,适当地提高有机营养水平可以减缓抑制效应。综合考虑CFU数量、种类丰富度、新种质资源、溶解无机磷细菌及产铁载体细菌的获得情况,采用1/50×R2A+1.0% NaCl+pH 8.5的培养基可以获得良好的分离效果。【结论】本研究结果获得了一批重度盐碱地细菌资源,也为其他盐碱地细菌新资源的分离培养提供了参考。

    Abstract:

    [Background] The saline-alkali land as a reserve arable resource is crucial to grain security. Plant growth-promoting rhizobacteria (PGPR) have been proved effective in improving saline-alkali land. [Objective] To explore the methods efficient in isolating PGPR and promote the development and utilization of PGPR in saline-alkali land. [Methods] The rhizosphere soil of soybean plants grown in the heavy saline-alkali land of Dongying City was collected for the isolation of bacteria by the dilution culture method. The effects of pH, organic nutrient level, and NaCl concentration on the isolation were evaluated. The strains capable of solubilizing inorganic phosphorus and producing siderophores were screened on functional plates. The purified isolates were identified by sequencing the 16S rRNA gene. [Results] A total of 443 isolates were successfully sequenced for the 16S rRNA gene, and they were identified as 96 species of 48 genera and 7 potential new species. Among these species, 28 and 13 were capable of solubilizing inorganic phosphorus and producing siderophores, respectively, and 7 species showed both functions. From points of bacterial richness and CFU count, NaCl concentration showed the greatest effect, followed by organic nutrient level, and then pH. Under the oligotrophic condition, NaCl at a concentration ≥3% significantly inhibited the growth of bacteria isolated from saline-alkali soil, which could be alleviated considerably by increasing the organic nutrient level. Considering the CFU count, species richness, new germplasm resources, inorganic phosphorus solubilizing, and siderophore production, a medium containing 1.0% NaCl, 1/50×R2A, and with pH 8.5 was proposed to isolate bacteria from saline-alkali soil. [Conclusion] We isolate a batch of bacterial resources from heavy saline-alkali land and provide methodological reference for isolating new bacterial resources from saline-alkali land in other areas.

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谭寿生,谭心许,李怡君,杨芾,韦善君,张晓霞. 重度盐碱地细菌的资源分离培养方法[J]. 微生物学通报, 2025, 52(7): 3255-3270

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  • 收稿日期:2024-11-14
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  • 录用日期:2025-03-09
  • 在线发布日期: 2025-07-21
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