降盐碱功能菌株的筛选、作用机制及应用效果评价

Screening, mechanisms of action, and application effect evaluation of salt-alkali reducing functional strains

  • 摘要:
    目的 筛选适宜于盐碱环境和具有降盐碱功能的菌株,解析其功能机制并评估其盐碱土壤修复效果。
    方法 通过滴定法和pH测定评价菌株降盐碱效果,并在不同NaCl浓度和pH梯度下测试菌株耐受性。此外,分析外源添加物(甘氨酸、谷氨酸、柠檬酸、甜菜碱和脯氨酸)与金属离子(K+、Ca2+、Mg2+、Fe2+和Mn2+)对菌株降盐碱效率的影响。在此基础上,进行菌株全基因组测序,挖掘相关功能基因,并通过土壤改良实验,测定pH、电导率、交换性钠百分比和全盐量等指标,评估菌株土壤修复应用潜力。最后,采用响应面法优化菌株发酵条件。
    结果 获得2株降盐碱细菌,编号为11和14,鉴定为沼泽微杆菌和维罗纳假单胞菌。在初始盐度为10条件下,菌株11和菌株14降盐率分别达到3.95%±0.24%和3.74%±0.16%;初始pH为9.0条件下,降碱率分别达到17.19%±0.45%和17.12%±0.40%,两菌株在1%~3% NaCl及pH 7~9下稳定生长。此外,柠檬酸和甜菜碱能够显著提升菌株11的降盐率,谷氨酸和柠檬酸能够显著提升菌株14的降盐率;而金属离子对两菌株降碱效果无显著影响。基因组分析表明,菌株11和菌株14分别包含59个和75个降盐碱相关基因,这些基因功能分类涵盖离子转运、渗透调节和有机酸代谢。土壤改良实验中,菌株11显著降低盐碱土壤交换性钠百分比和pH,菌株14显著降低电导率和全盐量,混合处理无协同增效。菌株11的最佳发酵条件为接种量3.46%、盐度9.83和pH 7.97;菌株14的最佳发酵条件为接种量2.09%、盐度3.19和pH 8.00。
    结论 菌株11可显著降低盐碱土壤的交换性钠百分比和pH;菌株14可显著降低土壤电导率和全盐量,在盐碱土壤生物修复领域具有良好应用前景。

     

    Abstract: Microbial remediation has been recognized as an environmentally friendly and cost-effective approach for improving saline-alkali soils. This study aimed to isolate bacterial strains capable of reducing salinity and alkalinity from saline-alkali soils, elucidate their functional mechanisms, and evaluate their remediation potential. Salt-alkali reduction efficiency was assessed by titration and pH measurement, and tolerance was tested across different NaCl concentrations and pH gradients. The effects of exogenous additives and metal ions on salt-alkali reduction efficiency were analyzed, whole-genome sequencing was performed to identify functional genes. Soil improvement experiments were conducted to evaluate remediation potential by measuring pH, electrical conductivity, exchangeable sodium percentage(ESP), and total salt content. Fermentation conditions were optimized using response surface methodology. Two salt-alkali-reducing bacterial strains, designated as 11 and 14, were identified as Microbacterium paludicola and Pseudomonas veronii, respectively. Under an initial salinity of 10, salt reduction rates reached 3.95%±0.24% and 3.74%±0.16%; under an initial pH of 9.0, alkali reduction rates were 17.19%±0.45% and 17.12%±0.40%, respectively. Both strains grew stably under 1%-3% NaCl and pH 7-9. Citric acid and betaine significantly enhanced the salt reduction rate of strain 11, while glutamic acid and citric acid significantly improved its alkali reduction rate. Metal ions had no significant effect on the alkali reduction of either strain. Genomic analysis revealed 59 and 75 salt-alkali-related genes in strains 11 and 14, respectively, spanning ion transport, osmotic regulation, and organic acid metabolism. In soil improvement experiments, strain 11 significantly reduced ESP and pH, while strain 14 significantly decreased electrical conductivity and total salt content; mixed inoculation showed no synergistic effect. Optimal fermentation conditions for strain 11 were an inoculation volume of 3.46%, salinity of 9.83, and pH of 7.97, with a verified OD600 of 0.668; for strain 14, inoculation volume of 2.09%, salinity of 3.19, and pH of 8.00, with a verified OD600 of 0.963. Strains 11 and 14 reduce soil salinity and alkalinity through distinct mechanisms of organic acid secretion and membrane transport-mediated ion uptake, respectively, and both demonstrate promising potential for the biological remediation of saline-alkali soils.

     

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