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微囊化墨西哥山达克酵母LPa14的等温微量热分析:对胶孢炭疽菌的释放及抗真菌活性

Isothermal microcalorimetry analysis of microencapsulated Yamadazyma mexicana LPa14: release and antifungal activity against Colletotrichum gloeosporioides.

作者信息

González-Gutiérrez Katia Nayely, Covarrubias-Rivera Leonardo, Ragazzo-Sánchez Juan Arturo, López-Cruz Rafael, Calderón-Santoyo Montserrat, Nayely Katia

机构信息

Laboratorio Integral de Investigación en Alimentos, Tecnológico Nacional de México, Instituto Tecnológico de Tepic, Av. Tecnológico 2595, Tepic, Nayarit, 63175, México.

出版信息

Arch Microbiol. 2025 Sep 16;207(11):264. doi: 10.1007/s00203-025-04470-5.

Abstract

Anthracnose caused by Colletotrichum is the most significant disease in avocado. Since its quiescent infection begins in the field, preharvest control is crucial. Applying microencapsulated Yamadazyma mexicana LPa14 in preharvest may prevent pathogen establishment and reduce production losses. However, the evaluation of the release and the antagonistic activity of microencapsulated yeasts is essential to ensure the efficacy of a bioformulation. Due to the limitations of traditional techniques, isothermal microcalorimetry (IMC) is proposed as an innovative, accurate, and sensitive technique for measuring the heat generated during biological processes. This work aimed to evaluate the release of microencapsulated Y. mexicana and its antifungal activity against Colletotrichum gloeosporioides from avocado fruit using IMC and traditional microbiological techniques. The intermediate release rate of the bioformulation was compared with low and high release rate treatments, and Pearson correlation was used to associate the release rates with the thermokinetic parameters. For the confrontation test, the thermogenic curves of yeast (1.14 × 10 and 3.16 × 10 cells/mL) and fungus were recorded, the co-culture curves were deconvoluted and the thermokinetic parameters: lag phase (λ), maximum growth rate (µ), total heat (Q), and time to peak (T) were obtained. The release rates were inversely correlated with Q. In vitro, C. gloeosporioides inhibition was yeast concentration-dependent, with no differences between fresh (56.78-68.24%) and microencapsulated (54.21-70.90%) yeast, as well as for the thermokinetic values. In the confrontation assay, at 3.16 × 10 cells/mL, the yeast and confrontation curves showed a similar pattern, with no significant differences between µ, λ phase, Q, and T, indicating complete fungal inhibition. Instead, at 1.14 × 10 cells/mL, the yeast and fungus growth is affected, indicating incomplete inhibition below this concentration. In conclusion, the IMC is suitable for in vitro testing, as it provides insights into the release dynamics and antifungal activity of microencapsulated biocontrol agents. This study is crucial for enhancing the efficacy and practical application of bioformulations for fungal diseases management.

摘要

由炭疽菌引起的炭疽病是鳄梨中最严重的病害。由于其潜伏感染始于田间,收获前的防治至关重要。在收获前施用微囊化的墨西哥山田酵母LPa14可以防止病原菌定殖并减少生产损失。然而,评估微囊化酵母的释放和拮抗活性对于确保生物制剂的功效至关重要。由于传统技术的局限性,等温微量热法(IMC)被提议作为一种用于测量生物过程中产生热量的创新、准确且灵敏的技术。这项工作旨在使用IMC和传统微生物技术评估微囊化墨西哥山田酵母的释放及其对鳄梨果实上炭疽菌的抗真菌活性。将生物制剂的中等释放速率与低释放速率和高释放速率处理进行比较,并使用皮尔逊相关性将释放速率与热动力学参数相关联。对于对峙试验,记录酵母(1.14×10和3.16×10个细胞/mL)和真菌的产热曲线,对共培养曲线进行反卷积并获得热动力学参数:延迟期(λ)、最大生长速率(µ)、总热量(Q)和达到峰值的时间(T)。释放速率与Q呈负相关。在体外,炭疽菌的抑制作用取决于酵母浓度,新鲜酵母(56.78 - 68.24%)和微囊化酵母(54.21 - 70.90%)之间以及热动力学值之间没有差异。在对峙试验中,在3.16×10个细胞/mL时,酵母和对峙曲线显示出相似的模式,µ、λ期、Q和T之间没有显著差异,表明真菌被完全抑制。相反,在1.14×10个细胞/mL时,酵母和真菌的生长受到影响,表明低于该浓度时抑制不完全。总之,IMC适用于体外测试,因为它提供了对微囊化生物防治剂释放动态和抗真菌活性的见解。这项研究对于提高生物制剂在真菌病害管理中的功效和实际应用至关重要。

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