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Bifidobacterium breve DSM 32583 and Limosilactobacillus fermentum CECT5716 postbiotics attenuate S. aureus and IL-33-induced Th2 responses 双歧杆菌(Bifidobacterium breve DSM 32583)和柠檬乳杆菌发酵体(Limosilactobacillus fermentum CECT5716)后益生菌可减轻金黄色葡萄球菌和 IL-33 诱导的 Th2 反应
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-21 DOI: 10.1016/j.micres.2024.127913
Yacine Amar , Jana Grube , Martin Köberle , Monika Schaubeck , Tilo Biedermann , Thomas Volz
Over the past decades, the prevalence of allergic diseases noticeably increased in industrialized countries. The Th2 immune response plays a central role in these pathologies and its modulation using pro-/postbiotics constitutes a promising approach to prevent or alleviate disease symptoms. The aim of this in vitro study, was to investigate the ability of human milk-derived Bifidobacterium breve DSM 32583 (Bb) and Limosilactobacillus fermentum CECT5716 (Lf), to modulate the Th2 induced responses. To this end, Th2 cells were generated by co-culturing of human naïve Th cells with monocyte-derived dendritic cells (moDCs) either stimulated with Staphylococcus aureus or IL-33. The immunomodulatory effects of pro-/postbiotic preparations of Bb and Lf on moDCs and Th2 cells were evaluated in terms of maturation markers expression and cytokines production. Remarkably, the tested strains induced the anti-inflammatory cytokine IL-10 in moDCs, in a strain-, dose- and viability-dependent manner with no significant upregulation of IL-12p70 nor CD83, CD86 or HLA-DR. Interestingly, Bb and Lf postbiotics were able to dampen the Th2/Th1 response induced upon S. aureus- or IL-33 stimulation. They were also able to synergistically induce IL-10 in moDCs and T cells, upon co-stimulation with LPS. Finally, we observed that live probiotics triggered a mild Th1 response that was attenuated in the presence of galacto-oligosaccharides. Altogether, Bb and Lf pro-/postbiotics exhibited remarkable immune regulatory effects on both moDCs and Th2 cells. Therefore, further in vivo studies should be considered to validate these findings and assess their ability to prevent allergy or alleviate its symptoms in affected patients.
过去几十年来,工业化国家的过敏性疾病发病率明显上升。Th2 免疫反应在这些病症中起着核心作用,而使用益生菌/后益生菌对其进行调节是预防或减轻疾病症状的一种很有前景的方法。这项体外研究的目的是调查人乳衍生的乳双歧杆菌 DSM 32583(Bb)和低硅乳杆菌发酵体 CECT5716(Lf)调节 Th2 诱导反应的能力。为此,通过用金黄色葡萄球菌或 IL-33 刺激人类幼稚 Th 细胞与单核细胞衍生树突状细胞(moDCs)共培养,产生 Th2 细胞。从成熟标志物表达和细胞因子产生的角度评估了 Bb 和 Lf 的前/后生物制剂对 moDCs 和 Th2 细胞的免疫调节作用。值得注意的是,所测试的菌株能诱导 moDCs 中的抗炎细胞因子 IL-10,其诱导方式与菌株、剂量和存活率有关,而 IL-12p70 或 CD83、CD86 或 HLA-DR 均无明显上调。有趣的是,Bb 和 Lf 后生物能抑制金黄色葡萄球菌或 IL-33 刺激诱导的 Th2/Th1 反应。在 LPS 的共同刺激下,它们还能协同诱导 moDCs 和 T 细胞中的 IL-10。最后,我们观察到活益生菌会引发轻微的 Th1 反应,而这种反应在半乳寡糖的存在下会减弱。总之,Bb 和 Lf 原/后益生菌对 moDCs 和 Th2 细胞都有显著的免疫调节作用。因此,应考虑进一步开展体内研究,以验证这些发现,并评估它们预防过敏或减轻过敏患者症状的能力。
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引用次数: 0
Arbuscular mycorrhizal fungi and exogenous Ca2+ application synergistically enhance salt and alkali resistance in perennial ryegrass through diverse adaptive strategies 丛枝菌根真菌和外源 Ca2+ 的应用通过不同的适应策略协同增强多年生黑麦草的抗盐抗碱能力
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-20 DOI: 10.1016/j.micres.2024.127906
Hongjian Wei , Wenyuan He , Xinjie Mao , Songkai Liao , Qi Wang , Zhihao Wang , Ming Tang , Tingying Xu , Hui Chen
The challenge of soil salinization and alkalization, with its significant impact on crop productivity, has raised growing concerns with global population growth and enhanced environmental degradation. Although arbuscular mycorrhizal fungi (AMF) and calcium ions (Ca2+) are known to enhance plant resistance to stress, their combined effects on perennial ryegrass’ tolerance to salt and alkali stress and the underlying mechanisms remain poorly understood. This study aimed to elucidate the roles of Arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis and exogenous Ca2+ application in molecular and physiological responses to salt-alkali stress. AM symbiosis and exogenous Ca2+ application enhanced antioxidant enzyme activity and non-enzymatic components, promoting reactive oxygen species (ROS) scavenging and reducing lipid peroxidation while alleviating oxidative damage induced by salt-alkali stress. Furthermore, they enhanced osmotic balance by increasing soluble sugar content (Proportion of contribution of the osmotic adjustment were 34∼38 % in shoots and 30∼37 % in roots) under salt stress and organic acid content (Proportion of contribution of the osmotic adjustment were 32∼36 % in shoots and 37∼42 % in roots) under alkali stress. Changes in organic solute and inorganic cation-anion contents contributed to ion balance, while hormonal regulation played a role in these protective mechanisms. Moreover, the protective mechanisms involved activation of Ca2+-mediated signaling pathways, regulation of salt-alkali stress-related genes (including LpNHX1 and LpSOS1), increased ATPase activity, elevated ATP levels, enhanced Na+ extrusion, improved K+ absorption capacity, and a reduced Na+/K+ ratio, all contributing to the protection of photosynthetic pigments and the enhancement of photosynthetic efficiency. Ultimately, the combined application of exogenous Ca2+ and AMF synergistically alleviated the inhibitory effects of salt-alkali stress on perennial ryegrass growth. This finding suggested that exogenous Ca2+ may participate in the colonization of perennial ryegrass plants by R. irregularis, while AM symbiosis may activate Ca2+ pathways. Consequently, the combined treatment of AM and Ca2+ is beneficial for enhancing plant regulatory mechanisms and increasing crop yield under salt-alkali stress.
随着全球人口增长和环境恶化加剧,土壤盐碱化的挑战及其对作物生产力的重大影响日益引起人们的关注。众所周知,丛枝菌根真菌(AMF)和钙离子(Ca2+)能增强植物的抗逆性,但它们对多年生黑麦草耐盐碱胁迫的综合影响及其内在机制仍鲜为人知。本研究旨在阐明丛枝菌根(AM)真菌根瘤菌(Rhizophagus irregularis)和外源 Ca2+ 在盐碱胁迫分子和生理反应中的作用。AM共生和外源Ca2+的应用增强了抗氧化酶活性和非酶成分,促进了活性氧(ROS)清除,降低了脂质过氧化,同时减轻了盐碱胁迫引起的氧化损伤。此外,在盐胁迫下,它们通过增加可溶性糖含量(对渗透调节的贡献比例在芽中为 34∼38%,在根中为 30∼37%)和有机酸含量(对渗透调节的贡献比例在芽中为 32∼36%,在根中为 37∼42%)来提高渗透平衡。有机溶质和无机阳离子-阴离子含量的变化有助于离子平衡,而激素调节在这些保护机制中也发挥了作用。此外,保护机制还包括激活 Ca2+ 介导的信号通路、调节盐碱胁迫相关基因(包括 LpNHX1 和 LpSOS1)、增加 ATP 酶活性、提高 ATP 水平、增强 Na+ 挤压、提高 K+ 吸收能力以及降低 Na+/K+ 比率,所有这些都有助于保护光合色素和提高光合效率。最终,外源 Ca2+ 和 AMF 的联合应用协同缓解了盐碱胁迫对多年生黑麦草生长的抑制作用。这一发现表明,外源 Ca2+ 可能参与了 R. irregularis 对多年生黑麦草植株的定殖,而 AM 共生可能激活 Ca2+ 途径。因此,在盐碱胁迫下,AM 和 Ca2+ 的联合处理有利于增强植物调节机制,提高作物产量。
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引用次数: 0
Halotolerant Enterobacter asburiae A103 isolated from the halophyte Salix linearistipularis: Genomic analysis and growth-promoting effects on Medicago sativa under alkali stress 从盐生植物 Salix linearistipularis 中分离出的耐卤肠杆菌 A103:基因组分析及在碱胁迫下对麦冬草的生长促进作用
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-19 DOI: 10.1016/j.micres.2024.127909
Yulin Li , Mengya Gao , Weiting Zhang , Yuchen Liu , Shanshan Wang , Huihui Zhang , Xiaoyan Li , Shuyu Yu , Lei Lu

Soil salinization negatively affects plant growth and threatens food security. Halotolerant plant growth-promoting bacteria (PGPB) can alleviate salt stress in plants via diverse mechanisms. In the present study, we isolated salt-tolerant bacteria with phosphate-solubilizing abilities from the rhizosphere of Salix linearistipularis, a halophyte distributed in saline-alkali soils. Strain A103 showed high phosphate solubilization activity and was identified as Enterobacter asburiae based on genome analysis. In addition, it can produce indole-3-acetic acid (IAA), siderophores, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase. Genome mining has also revealed the presence of several functional genes involved in the promotion of plant growth. Inoculation with A103 markedly improved alfalfa growth in the presence of 100 mM NaHCO3. Under alkali stress, the shoot and root dry weights after bacterial inoculation improved by 42.9 % and 21.9 %, respectively. Meanwhile, there was a 35.9–37.1 % increase in the shoot and root lengths after treatment with A103 compared to the NaHCO3-treated group. Soluble sugar content, peroxidase and catalase activities increased in A103-inoculated alfalfa under alkaline stress. A significant decrease in the malondialdehyde content was observed after treatment with strain A103. Metabolomic analysis indicated that strain A103 positively regulated alkali tolerance in alfalfa through the accumulation of metabolites, such as homocarnosine, panthenol, and sorbitol, which could reduce oxidative damage and act as osmolytes. These results suggest that halophytes are valuable resources for bioprospecting halotolerant beneficial bacteria and that the application of halotolerant growth-promoting bacteria is a natural and efficient strategy for developing sustainable agriculture.

土壤盐碱化对植物生长造成负面影响,并威胁到粮食安全。耐盐植物生长促进菌(PGPB)可通过多种机制缓解植物的盐胁迫。在本研究中,我们从分布于盐碱地的盐生植物 Salix linearistipularis 的根瘤中分离到了具有磷酸盐溶解能力的耐盐细菌。菌株 A103 表现出很高的磷酸盐溶解活性,根据基因组分析被鉴定为 Enterobacter asburiae。此外,它还能产生吲哚-3-乙酸(IAA)、苷元和 1-氨基环丙烷-1-羧酸(ACC)脱氨酶。基因组挖掘还发现了几个参与促进植物生长的功能基因。在 100 mM NaHCO3 存在的情况下,接种 A103 能明显改善紫花苜蓿的生长。在碱胁迫条件下,接种细菌后的芽干重和根干重分别提高了 42.9% 和 21.9%。同时,与 NaHCO3 处理组相比,A103 处理组的芽和根长度增加了 35.9%-37.1%。在碱性胁迫下,接种 A103 的紫花苜蓿的可溶性糖含量、过氧化物酶和过氧化氢酶活性都有所增加。经菌株 A103 处理后,丙二醛含量明显下降。代谢组学分析表明,菌株 A103 通过积累代谢物(如高肌肽、泛醇和山梨醇)积极调节苜蓿的耐碱性,这些代谢物可减少氧化损伤并起到渗透溶解作用。这些结果表明,卤植物是生物勘探耐碱有益菌的宝贵资源,应用耐碱生长促进菌是发展可持续农业的一种自然而有效的策略。
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引用次数: 0
Plant growth-promoting microorganisms drive K strategists through deterministic processes to alleviate biological stress caused by Fusarium oxysporum 植物生长促进微生物通过决定性过程驱动钾策略因子,以减轻镰刀菌造成的生物胁迫
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-18 DOI: 10.1016/j.micres.2024.127911
Liangyang Mao , Bo Yin , Zeming Ye , Jie Kang , Rui Sun , Zhenchao Wu , Jingping Ge , Wenxiang Ping

Soybean root rot, caused by soil-borne pathogens such as Fusarium oxysporum, frequently occurs in Northeast China and leads to a decline in soil health and becoming a bottleneck for soybean yield in the region. To address this issue, applying beneficial microorganisms and altering soil microbial community structure have become effective strategies. In this study, the 90-day soybean pot experiment was conducted to explore the assembly process and life strategy selection of bacterial communities in the rhizosphere of healthy (inoculated with Funneliformis mosseae, F group and treated with Pseudomonas putida, P group) and diseased (inoculated with F. oxysporum, O group) soybean plants, as well as the recovery effect of beneficial microorganisms on soil-borne diseases (combined treatments OP and OF). Results indicated that in healthy soils (P and F), microbial community assembly process in the soybean rhizosphere was entirely governed by heterogeneous selection (HeS, 100 %). However, inoculated with P. putida (OP) was primarily driven by stochastic processes (HeS 40 %, dispersal limitation (DL) 60 %), and the F. mosseae treatment (OF) predominantly followed a deterministic process (HeS 89 %, DL 11 %) in diseased soils. Inoculation of plant growth-promoting microorganisms (PGPMs) in diseased soil drove the life strategy of the rhizosphere bacterial community from r- to K-strategy, evident from the lower rRNA operon (rrn) copy numbers (O 3.7, OP 2.1, OF 2.3), higher G+ to G- ratios (O 0.47, OP 0.58, OF 0.57), and a higher abundance of oligotrophs (O 50 %, OP 53 %, OF 54 %). In healthy (P and F) and diseased (O, OP, OF) rhizosphere soils, OTU820, OTU6142, and OTU8841 under the K-strategy, and OTU6032 and OTU6917 under the r-strategy, which served as keystone species, had a significant promoting relationship with plant biomass and defense capabilities ( p <0.05). Additionally, inoculation of PGPMs improved autotoxin degradation and positively correlated with bacterial life strategies in both healthy and diseased soils (P, F, OP and OF) ( p <0.05). These findings enhance our understanding of soil-microbe interactions and offer new insights and precise control measures for soybean disease management and soil environment remediation.

大豆根腐病是由镰刀菌等土传病原菌引起的,在中国东北地区经常发生,导致土壤健康状况下降,成为该地区大豆产量的瓶颈。为解决这一问题,施用有益微生物和改变土壤微生物群落结构已成为有效的策略。本研究通过为期 90 天的大豆盆栽实验,探讨了健康大豆植株(接种 Funneliformis mosseae,F 组,用 Pseudomonas putida 处理,P 组)和患病大豆植株(接种 F. oxysporum,O 组)根瘤菌群落的组装过程和生命策略选择,以及有益微生物对土传病害(OP 和 OF 组合处理)的恢复效果。结果表明,在健康土壤(P 和 F)中,大豆根圈微生物群落的形成过程完全受异质性选择(HeS,100%)的控制。然而,在病害土壤中,接种 P. putida(OP)主要由随机过程驱动(HeS 40 %,扩散限制 (DL) 60 %),而 F. mosseae 处理(OF)则主要遵循确定性过程(HeS 89 %,DL 11 %)。在病害土壤中接种促进植物生长的微生物(PGPMs)后,根瘤菌群落的生活战略从 r 战略转变为 K 战略,这从较低的 rRNA 操作子(rrn)拷贝数(O 3.7、OP 2.1、OF 2.3)、较高的 G+ 与 G- 比率(O 0.47、OP 0.58、OF 0.57)和较高的寡养菌丰度(O 50%、OP 53%、OF 54%)中可以看出。在健康(P 和 F)和病害(O、OP、OF)根瘤土壤中,K 策略下的 OTU820、OTU6142 和 OTU8841,以及 r 策略下的 OTU6032 和 OTU6917 作为关键种,与植物的生物量和防御能力有显著的促进关系 ( p <0.05)。此外,接种 PGPMs 可改善自毒素降解,并与健康土壤和病害土壤中的细菌生活策略(P、F、OP 和 OF)呈正相关(p <0.05)。这些发现加深了我们对土壤-微生物相互作用的理解,为大豆病害管理和土壤环境修复提供了新的见解和精确的控制措施。
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引用次数: 0
PGPR inoculants journey from lab to land: Challenges and limitations PGPR 接种剂从实验室到土地的过程:挑战与局限
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-12 DOI: 10.1016/j.micres.2024.127910
Rishu Thakur , Hena Dhar , Supriya Mathew , Arvind Gulati
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引用次数: 0
Unveiling the genetic basis and metabolic rewiring behind the galactose-positive phenotype in a Streptococcus thermophilus mutant 揭示嗜热链球菌突变体半乳糖阳性表型背后的遗传基础和代谢线路重构
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-11 DOI: 10.1016/j.micres.2024.127894
Haimin Hu , Qingting Peng , Jiahui Tai , Wenhui Lu , Jinhui Liu , Tong Dan

Streptococcus thermophilus (S. thermophilus) is a widely used starter culture in dairy fermentation, but most strains are galactose-negative and only metabolize glucose from lactose hydrolysis. In this study, we aimed to uncover the mechanisms underlying the acquisition of a stable galactose-positive (Gal+) phenotype in a mutant strain of S. thermophilus IMAU10636. By treating the wild-type strain with the mutagenic agent N-methyl-N-nitro-N-nitrosoguanidine, we successfully isolated a Gal+ mutant, S. thermophilus IMAU10636Y. Comparative enzyme activity assays revealed that the mutant exhibited higher β-galactosidase and galactokinase activities, but lower glucokinase and pyruvate kinase activities compared to the wild-type. High-performance liquid chromatography analysis confirmed the mutant’s enhanced ability to utilize lactose and galactose, leading to increased glucose secretion. Integrated genome and transcriptomics analyses provided deeper insights into the underlying genetic and metabolic mechanisms. We found that the metabolism regulatory network of the glycolysis / Leloir pathway was altered in the mutant, possibly due to the upregulation of the gene expression in the galR-galK intergenic region. This likely led to increased RNA polymerase binding and transcription of the gal operon, ultimately promoting the Gal+ phenotype. Additionally, we identified a mutation in the scrR gene, encoding a LacI family transcriptional repressor, which also contributed to the Gal+ phenotype. These findings offer new perspectives on the metabolic rewiring and regulatory mechanisms that enable S. thermophilus to acquire the ability to metabolize galactose. This knowledge can inform strategies for engineering and selecting Gal+ strains with desirable fermentation characteristics for dairy applications.

嗜热链球菌(S. thermophilus)是乳制品发酵中广泛使用的启动培养物,但大多数菌株都是半乳糖阴性的,只能代谢乳糖水解产生的葡萄糖。在本研究中,我们旨在揭示嗜热杆菌突变菌株 IMAU10636 获得稳定的半乳糖阳性(Gal+)表型的机制。通过用诱变剂 N-甲基-N-亚硝基-N-亚硝基胍处理野生型菌株,我们成功分离出了 Gal+ 突变体嗜热菌 IMAU10636Y。酶活性比较试验显示,与野生型相比,突变体具有更高的β-半乳糖苷酶和半乳激酶活性,但葡萄糖激酶和丙酮酸激酶活性较低。高效液相色谱分析证实,突变体利用乳糖和半乳糖的能力增强,导致葡萄糖分泌增加。基因组学和转录组学的综合分析使我们对潜在的遗传和代谢机制有了更深入的了解。我们发现,突变体中糖酵解/Leloir 途径的代谢调控网络发生了改变,这可能是由于 galR-galK 基因间区域的基因表达上调所致。这可能导致 RNA 聚合酶的结合和 gal 操作子的转录增加,最终促进了 Gal+ 表型的形成。此外,我们还发现编码 LacI 家族转录抑制因子的 scrR 基因发生了突变,这也导致了 Gal+ 表型的形成。这些发现为嗜热菌获得半乳糖代谢能力的代谢重构和调控机制提供了新的视角。这些知识可为乳制品应用中具有理想发酵特性的 Gal+ 菌株的工程和选择策略提供参考。
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引用次数: 0
Elaborating the multifarious role of PGPB for sustainable food security under changing climate conditions 阐述植物基因工程在不断变化的气候条件下促进可持续粮食安全的多重作用
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-07 DOI: 10.1016/j.micres.2024.127895
Margi Patel , Shaikhul Islam , Bernard R. Glick , Shobhit Raj Vimal , Sachin Ashok Bhor , Matteo Bernardi , Fatema Tuj Johora , Ashish Patel , Sergio de los Santos Villalobos

Changing climate creates a challenge to agricultural sustainability and food security by changing patterns of parameters like increased UV radiation, rising temperature, altered precipitation patterns, and higher occurrence of extreme weather incidents. Plants are vulnerable to different abiotic stresses such as waterlogging, salinity, heat, cold, and drought in their natural environments. The prevailing agricultural management practices play a major role in the alteration of the Earth's climate by causing biodiversity loss, soil degradation through chemical and physical degradation, and pollution of water bodies. The extreme usage of pesticides and fertilizers leads to climate change by releasing greenhouse gases (GHGs) and depositing toxic substances in the soil. At present, there is an urgent need to address these abiotic stresses to achieve sustainable growth in agricultural production and fulfill the rising global food demand. Several types of bacteria that are linked with plants can increase plant resistance to stress and lessen the negative effects of environmental challenges. This review aims to explore the environmentally friendly capabilities and prospects of multi-trait plant growth-promoting bacteria (PGPB) in the alleviation of detrimental impacts of harsh environmental conditions on plants.

不断变化的气候改变了紫外线辐射增加、气温升高、降水模式改变和极端天气事件增多等参数模式,给农业可持续性和粮食安全带来了挑战。植物在自然环境中很容易受到不同的非生物胁迫,如水涝、盐碱、高温、寒冷和干旱。普遍的农业管理方法导致生物多样性丧失、土壤因化学和物理退化而退化以及水体污染,从而对地球气候的改变起到了重要作用。杀虫剂和化肥的大量使用会释放温室气体,并在土壤中沉积有毒物质,从而导致气候变化。目前,迫切需要解决这些非生物压力,以实现农业生产的可持续增长,满足全球不断增长的粮食需求。与植物相关的几种细菌可以增强植物的抗逆性,减轻环境挑战的负面影响。本综述旨在探讨多性状植物生长促进细菌(PGPB)在减轻恶劣环境条件对植物的不利影响方面的环境友好能力和前景。
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引用次数: 0
KPC variants conferring resistance to ceftazidime-avibactam in Pseudomonas aeruginosa strains 铜绿假单胞菌株中对头孢他啶-阿维巴坦产生耐药性的 KPC 变体
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-07 DOI: 10.1016/j.micres.2024.127893
Yanyan Hu , Weiyi Shen , Di Lin , Yuchen Wu , Yanyan Zhang , Hongwei Zhou , Rong Zhang

Background

This study aimed to characterize three KPC variants (KPC-33, KPC-100, and KPC-201) obtained from a clinical isolate of Pseudomonas aeruginosa (#700), along with two induced strains C109 and C108.

Methods

Genomic DNAs of #700 (ST235), C109 (ST463), and C108 (ST1076) were sequenced using Illumina and Oxford Nanopore technologies. The transferability and stability of the plasmid was assessed through conjugation experiments and plasmid stability experiments, respectively. Minimum inhibitory concentrations of bacterial strains were determined using broth microdilution methods. In vitro induction was performed using ceftazidime-avibactam (CZA) at concentrations of 6/4 µg/ml. Linear genomic alignments were visualized using Easyfig, and protein structure modeling of the novel KPC variant (KPC-201) was conducted using PyMol.

Results

The plasmids carrying the KPC variants in the three CZA-resistant strains (C109, C108, and #700) had sizes of 39,251 bp (KPC-100), 394,978 bp (KPC-201), and 48,994 bp (KPC-33). All three plasmids belonged to the IncP-like incompatibility (Inc) groups, and the plasmid exhibited relatively high plasmid stability, KPC-33 and KPC-201-harboring plasmids were successfully transferred to the recipient strain P. aeruginosa PAO1rifR. The genetic environments of the three blaKPC genes differed from each other. The mobile elements of the three blaKPC genes were as follows, TnAS1-IS26-ΔISKpn27-blaKPC-33-ISKpn6-IS26, IS6-ΔISKpn27-blaKPC-100-ISKpn6-IS26-Tn3-IS26, and IS6100-ISKpn27-blaKPC-201-ISKpn6-TnAS1. Notably, the length of ΔISKpn27 upstream of the blaKPC-33 and blaKPC-100 genes were remarkably short, measuring 114 bp and 56 bp, respectively, deviating significantly from typical lengths associated with ISKpn27 elements. Moreover, the novel KPC variant, KPC-201, featured a deletion of amino acids LDR at positions 161–163 in KPC-3, resulting in a looser pocket structure contributing to its avibactam resistance.

Conclusions

KPC-201, identified as a novel KPC variant, exhibits resistance to CZA. The presence of multiple mobile elements surrounding the blaKPC-variant genes on stable plasmids is concerning. Urgent preventive measures are crucial to curb its dissemination in clinical settings.

背景本研究旨在鉴定从铜绿假单胞菌临床分离株(700 号)以及两株诱导株 C109 和 C108 中获得的三个 KPC 变体(KPC-33、KPC-100 和 KPC-201)的特征。方法使用 Illumina 和 Oxford Nanopore 技术对 700 号(ST235)、C109(ST463)和 C108(ST1076)的基因组 DNA 进行测序。质粒的转移性和稳定性分别通过共轭实验和质粒稳定性实验进行了评估。使用肉汤微稀释法测定细菌菌株的最小抑菌浓度。使用头孢唑肟-阿维巴坦(CZA)进行体外诱导,浓度为 6/4 µg/ml。结果三个耐 CZA 菌株(C109、C108 和 #700)中携带 KPC 变体的质粒大小分别为 39,251 bp(KPC-100)、394,978 bp(KPC-201)和 48,994 bp(KPC-33)。这三个质粒都属于类 IncP 不相容(Inc)组,质粒表现出较高的稳定性,KPC-33 和 KPC-201 载体质粒被成功转入受体菌株铜绿微囊藻 PAO1rifR。三种 blaKPC 基因的遗传环境各不相同。三个 blaKPC 基因的移动元件如下:TnAS1-IS26-ΔISKpn27-blaKPC-33-ISKpn6-IS26、IS6-ΔISKpn27-blaKPC-100-ISKpn6-IS26-Tn3-IS26 和 IS6100-ISKpn27-blaKPC-201-ISKpn6-TnAS1。值得注意的是,blaKPC-33 和 blaKPC-100 基因上游的 ΔISKpn27 长度非常短,分别为 114 bp 和 56 bp,与 ISKpn27 元件的典型长度明显不同。此外,新型 KPC 变异株 KPC-201 在 KPC-3 的第 161-163 位缺失了 LDR 氨基酸,导致口袋结构更松散,从而产生了阿维菌素耐药性。稳定质粒上的 blaKPC 变异基因周围存在多个移动元件,这令人担忧。采取紧急预防措施遏制其在临床环境中的传播至关重要。
{"title":"KPC variants conferring resistance to ceftazidime-avibactam in Pseudomonas aeruginosa strains","authors":"Yanyan Hu ,&nbsp;Weiyi Shen ,&nbsp;Di Lin ,&nbsp;Yuchen Wu ,&nbsp;Yanyan Zhang ,&nbsp;Hongwei Zhou ,&nbsp;Rong Zhang","doi":"10.1016/j.micres.2024.127893","DOIUrl":"10.1016/j.micres.2024.127893","url":null,"abstract":"<div><h3>Background</h3><p>This study aimed to characterize three KPC variants (KPC-33, KPC-100, and KPC-201) obtained from a clinical isolate of <em>Pseudomonas aeruginosa</em> (#700), along with two induced strains C109 and C108.</p></div><div><h3>Methods</h3><p>Genomic DNAs of #700 (ST235), C109 (ST463), and C108 (ST1076) were sequenced using Illumina and Oxford Nanopore technologies. The transferability and stability of the plasmid was assessed through conjugation experiments and plasmid stability experiments, respectively. Minimum inhibitory concentrations of bacterial strains were determined using broth microdilution methods. <em>In vitro</em> induction was performed using ceftazidime-avibactam (CZA) at concentrations of 6/4 µg/ml. Linear genomic alignments were visualized using Easyfig, and protein structure modeling of the novel KPC variant (KPC-201) was conducted using PyMol.</p></div><div><h3>Results</h3><p>The plasmids carrying the KPC variants in the three CZA-resistant strains (C109, C108, and #700) had sizes of 39,251 bp (KPC-100), 394,978 bp (KPC-201), and 48,994 bp (KPC-33). All three plasmids belonged to the IncP-like incompatibility (Inc) groups, and the plasmid exhibited relatively high plasmid stability, KPC-33 and KPC-201-harboring plasmids were successfully transferred to the recipient strain <em>P. aeruginosa</em> PAO1<sup>rifR</sup>. The genetic environments of the three <em>bla</em><sub>KPC</sub> genes differed from each other. The mobile elements of the three <em>bla</em><sub>KPC</sub> genes were as follows, Tn<em>AS1</em>-IS<em>26</em>-ΔIS<em>Kpn27</em>-<em>bla</em><sub>KPC-33</sub>-IS<em>Kpn6</em>-IS<em>26</em>, IS<em>6</em>-ΔIS<em>Kpn27</em>-<em>bla</em><sub>KPC-100</sub>-IS<em>Kpn6</em>-IS<em>26-</em>Tn<em>3-</em>IS<em>26</em>, and IS<em>6100</em>-IS<em>Kpn27-bla</em><sub>KPC-</sub>201-IS<em>Kpn6</em>-Tn<em>AS1</em>. Notably, the length of ΔIS<em>Kpn27</em> upstream of the <em>bla</em><sub>KPC-33</sub> and <em>bla</em><sub>KPC-100</sub> genes were remarkably short, measuring 114 bp and 56 bp, respectively, deviating significantly from typical lengths associated with IS<em>Kpn27</em> elements. Moreover, the novel KPC variant, KPC-201, featured a deletion of amino acids LDR at positions 161–163 in KPC-3, resulting in a looser pocket structure contributing to its avibactam resistance.</p></div><div><h3>Conclusions</h3><p>KPC-201, identified as a novel KPC variant, exhibits resistance to CZA. The presence of multiple mobile elements surrounding the <em>bla</em><sub>KPC-variant</sub> genes on stable plasmids is concerning. Urgent preventive measures are crucial to curb its dissemination in clinical settings.</p></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"289 ","pages":"Article 127893"},"PeriodicalIF":6.1,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0944501324002945/pdfft?md5=8976363dad2e8b909be56f4372a0acf4&pid=1-s2.0-S0944501324002945-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142164225","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Conjugative transmission of virulence plasmid in Klebsiella pneumoniae mediated by a novel IncN-like plasmid 肺炎克雷伯菌中由新型 IncN 样质粒介导的毒力质粒的共轭传播
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-05 DOI: 10.1016/j.micres.2024.127896
Qi Xu , Miaomiao Xie , Xuemei Yang , Xiaoxuan Liu , Lianwei Ye , Kaichao Chen , Edward Wai-Chi Chan , Sheng Chen

Klebsiella pneumoniae (Kp) is increasingly recognized as a reservoir for a range of antibiotic resistance genes and a pathogen that frequently causes severe infections in both hospital and community settings. In this study, we have identified a novel mechanism of conjugative transfer of a non-conjugative virulence plasmid through the formation of a fusion plasmid between the virulence plasmid and a novel 59,162 bp IncN- plasmid. This plasmid was found to be a multidrug-resistance (MDR) plasmid and carried a T4SS cluster, which greatly facilitated the efficient horizontal transfer of the fusion plasmid between Kp strains. The fused virulence plasmid conferred the resistance of serum killing and macrophage phagocytosis to the transconjugants. Importantly, this plasmid was shown to be essential for Kp virulence in a mouse model. Mechanistic analysis revealed that the virulence factors encoded by this virulence plasmid contributed to resistance to in vivo clearance and induced a high level of proinflammatory cytokine IL-1β, which acts as an inducer for more neutrophil recruitment. The transmission of the fusion plasmid in Kp has the potential to convert it into both MDR and hypervirulent Kp, accelerating its evolution, and posing a serious threat to human health. The findings of this study provide new insights into the rapid evolution of MDR and hypervirulent Kp in recent years.

肺炎克雷伯菌(Kp)越来越被认为是一系列抗生素耐药基因的贮藏库,也是一种在医院和社区环境中经常引起严重感染的病原体。在这项研究中,我们发现了一种非共轭型毒力质粒的新型共轭转移机制,即毒力质粒与 59,162 bp 的新型 IncN- 质粒之间形成融合质粒。研究发现,这种质粒是一种具有多重耐药性(MDR)的质粒,并携带一个 T4SS 簇,这极大地促进了融合质粒在 Kp 菌株之间的高效水平转移。融合后的毒力质粒使转染株具有抗血清杀灭和巨噬细胞吞噬的能力。重要的是,在小鼠模型中,该质粒被证明对 Kp 的毒力至关重要。机理分析表明,该毒力质粒编码的毒力因子有助于抵抗体内清除,并诱导高水平的促炎细胞因子IL-1β,从而诱导更多的中性粒细胞募集。Kp中融合质粒的传播有可能使其转化为MDR和高病毒性Kp,加速其进化,对人类健康构成严重威胁。本研究的发现为近年来 MDR 和高病毒性 Kp 的快速进化提供了新的视角。
{"title":"Conjugative transmission of virulence plasmid in Klebsiella pneumoniae mediated by a novel IncN-like plasmid","authors":"Qi Xu ,&nbsp;Miaomiao Xie ,&nbsp;Xuemei Yang ,&nbsp;Xiaoxuan Liu ,&nbsp;Lianwei Ye ,&nbsp;Kaichao Chen ,&nbsp;Edward Wai-Chi Chan ,&nbsp;Sheng Chen","doi":"10.1016/j.micres.2024.127896","DOIUrl":"10.1016/j.micres.2024.127896","url":null,"abstract":"<div><p><em>Klebsiella pneumoniae</em> (<em>Kp</em>) is increasingly recognized as a reservoir for a range of antibiotic resistance genes and a pathogen that frequently causes severe infections in both hospital and community settings. In this study, we have identified a novel mechanism of conjugative transfer of a non-conjugative virulence plasmid through the formation of a fusion plasmid between the virulence plasmid and a novel 59,162 bp IncN- plasmid. This plasmid was found to be a multidrug-resistance (MDR) plasmid and carried a T4SS cluster, which greatly facilitated the efficient horizontal transfer of the fusion plasmid between <em>Kp</em> strains. The fused virulence plasmid conferred the resistance of serum killing and macrophage phagocytosis to the transconjugants. Importantly, this plasmid was shown to be essential for <em>Kp</em> virulence in a mouse model. Mechanistic analysis revealed that the virulence factors encoded by this virulence plasmid contributed to resistance to <em>in vivo</em> clearance and induced a high level of proinflammatory cytokine IL-1β, which acts as an inducer for more neutrophil recruitment. The transmission of the fusion plasmid in <em>Kp</em> has the potential to convert it into both MDR and hypervirulent <em>Kp</em>, accelerating its evolution, and posing a serious threat to human health. The findings of this study provide new insights into the rapid evolution of MDR and hypervirulent <em>Kp</em> in recent years.</p></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"289 ","pages":"Article 127896"},"PeriodicalIF":6.1,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142164245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Melatonin alleviates apple replant disease by regulating the endophytic microbiome of roots and phloridzin accumulation 褪黑素通过调节根部的内生微生物群和phloridzin的积累来减轻苹果移栽病害。
IF 6.1 1区 生物学 Q1 MICROBIOLOGY Pub Date : 2024-09-04 DOI: 10.1016/j.micres.2024.127897
Changqing Ma , Peihua Du , Yang Cao , Huaite Liu , Lisong Ma , Bowen Liang

Melatonin administration is an environmentally effective strategy to mitigate apple replant disease (ARD), but its mechanism of action is unknown. This study investigated the protective effect of melatonin on ARD and the underlying mechanism. In field experiments, melatonin significantly reduced phloridzin levels in apple roots and rhizosphere soil. A correlation analysis indicated that a potential antagonistic interaction between melatonin and phloridzin was crucial for improving soil physicochemical properties, increasing the diversity of endophytic bacterial communities in roots of apple seedlings, and promoting mineral element absorption by the plants. Melatonin also reduced the abundance of Fusarium in roots. The ability of melatonin to reduce phloridzin levels both in soil and in plants was also demonstrated in a pot experiment. Azovibrio were specifically recruited in response to melatonin and their abundance was negatively correlated with phloridzin levels. Fusarium species that have a negative impact on plant growth were also inhibited by melatonin. Our results show that melatonin improves the rhizosphere environment as well as the structure of the endophytic microbiota community, by reducing phloridzin levels in rhizosphere soil and roots. These regulatory effects of melatonin support its use to improve the physiological state of plants under ARD conditions and thereby overcome the barriers of perennial cropping systems.

施用褪黑激素是减轻苹果再植病(ARD)的一种环境有效策略,但其作用机制尚不清楚。本研究调查了褪黑激素对苹果再植病(ARD)的保护作用及其机理。在田间试验中,褪黑激素显著降低了苹果根部和根瘤土壤中的氯唑嗪含量。相关性分析表明,褪黑素与披碱草素之间潜在的拮抗作用对于改善土壤理化性质、增加苹果幼苗根部内生细菌群落的多样性以及促进植物对矿质元素的吸收至关重要。褪黑素还能减少根部镰刀菌的数量。盆栽实验也证明,褪黑激素能够降低土壤和植物中的杀螟丹含量。Azovibrio 对褪黑激素有特异性反应,它们的数量与 phloridzin 的水平呈负相关。褪黑激素还能抑制对植物生长有负面影响的镰刀菌。我们的研究结果表明,褪黑激素通过降低根瘤菌圈土壤和根部的氯唑嗪水平,改善了根瘤菌圈环境以及内生微生物群落的结构。褪黑激素的这些调节作用支持利用它来改善 ARD 条件下植物的生理状态,从而克服多年生耕作系统的障碍。
{"title":"Melatonin alleviates apple replant disease by regulating the endophytic microbiome of roots and phloridzin accumulation","authors":"Changqing Ma ,&nbsp;Peihua Du ,&nbsp;Yang Cao ,&nbsp;Huaite Liu ,&nbsp;Lisong Ma ,&nbsp;Bowen Liang","doi":"10.1016/j.micres.2024.127897","DOIUrl":"10.1016/j.micres.2024.127897","url":null,"abstract":"<div><p>Melatonin administration is an environmentally effective strategy to mitigate apple replant disease (ARD), but its mechanism of action is unknown. This study investigated the protective effect of melatonin on ARD and the underlying mechanism. In field experiments, melatonin significantly reduced phloridzin levels in apple roots and rhizosphere soil. A correlation analysis indicated that a potential antagonistic interaction between melatonin and phloridzin was crucial for improving soil physicochemical properties, increasing the diversity of endophytic bacterial communities in roots of apple seedlings, and promoting mineral element absorption by the plants. Melatonin also reduced the abundance of <em>Fusarium</em> in roots. The ability of melatonin to reduce phloridzin levels both in soil and in plants was also demonstrated in a pot experiment. <em>Azovibrio</em> were specifically recruited in response to melatonin and their abundance was negatively correlated with phloridzin levels. <em>Fusarium</em> species that have a negative impact on plant growth were also inhibited by melatonin. Our results show that melatonin improves the rhizosphere environment as well as the structure of the endophytic microbiota community, by reducing phloridzin levels in rhizosphere soil and roots. These regulatory effects of melatonin support its use to improve the physiological state of plants under ARD conditions and thereby overcome the barriers of perennial cropping systems.</p></div>","PeriodicalId":18564,"journal":{"name":"Microbiological research","volume":"289 ","pages":"Article 127897"},"PeriodicalIF":6.1,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142145962","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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