白色念珠菌菌丝诱导：优化培养基参数以加速GlcNAc增强菌丝生长。

IF 1.9 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Journal of microbiological methods Pub Date : 2025-09-28 DOI:10.1016/j.mimet.2025.107282

Rachana A. , Atheena P.V. , Allan J. Britto , Uttara Chakraborty , Ritu Raval

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引用次数: 0

摘要

菌丝形成是白色念珠菌的一个关键毒力特征，在念珠菌病的发生和发展中起着至关重要的作用。因此，它仍然是抗真菌药物靶点研究的一个重要领域。传统的菌丝诱导培养基通常含有多种成分，使其制备复杂。在这项研究中，我们优化了一种简单且具有成本效益的培养基，其成分最少，可诱导菌丝形成。在不同组合中，MF8与蛋白胨0.16 %、葡萄糖0.4 %和牛血清白蛋白0.25 %的组合均能产生中等程度的菌丝。进一步添加n -乙酰氨基葡萄糖（GlcNAc）可以促进菌丝的形成，使菌丝生长成团。为了确定GlcNAc和mgso4的不同组合对菌丝形成的影响，我们采用了JMP软件的实验设计（DOE）方法。GlcNAc对菌丝诱导有显著影响（p 2 = 0.26），MgSO4对菌丝诱导不显著（p > 0.05）。在Run 20条件下（30 mM GlcNAc, 1 mM MgSO₄）进一步分析基因表达和pH，发现优化培养基中菌丝特异性基因菌丝壁蛋白1（HWP1）和菌丝特异性G1 cyclin 1（HGC1）的转录激活显著。值得注意的是，在4 h时，HWP1的表达增加了100倍，在6 h时增加了85倍

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Hyphal induction in Candida albicans: Optimizing medium parameters to accelerate hyphal growth enhanced by GlcNAc

Hyphal formation is a key virulence trait in Candida albicans, playing a crucial role in the development and progression of candidiasis. As a result, it remains a significant area of interest for antifungal drug target studies. Conventional hyphal induction medium typically contain multiple components, making them complex to prepare. In this study, we have optimized a simple and cost-effective medium with minimal components that induces hyphal formation. Among different combination, MF8 with Peptone 0.16 %, Dextrose 0.4 % and BSA 0.25 % caused moderate hyphal formation. Further addition of N-acetylglucosamine (GlcNAc) enhanced hyphal formation and caused clumped mycelial growth. To determine the different combinations of GlcNAc and MgSO₄ that cause hyphal formation, we employed a Design of Experiment (DOE) approach using JMP software. GlcNAc was found to be a significant component in hyphal induction (p < 0.05, R² = 0.26) and MgSO₄ was not significant (p > 0.05). Further gene expression and pH was analysed under Run 20 conditions (30 mM GlcNAc, 1 mM MgSO₄) which revealed significant transcriptional activation of hyphal-specific genes hyphal wall protein 1(HWP1) and Hypha-specific G1 cyclin 1(HGC1) in the optimized medium. Notably, HWP1 expression increased 100-fold at 4 h and 85-fold at 6 h (p < 0.05), while HGC1 showed a 7.09-fold and 6.07-fold upregulation at the same time points (p < 0.05). Additionally, there was an initial shift toward alkalinity at 1 and 2 h time point and a drop to acidic pH at 4 h and 6 h time point suggesting hyphal formation and upregulation of hyphal specific genes induced with the lowering of pH in medium.

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来源期刊

Journal of microbiological methods 生物-生化研究方法

CiteScore

4.30

自引率

4.50%

发文量

151

审稿时长

29 days

期刊介绍： The Journal of Microbiological Methods publishes scholarly and original articles, notes and review articles. These articles must include novel and/or state-of-the-art methods, or significant improvements to existing methods. Novel and innovative applications of current methods that are validated and useful will also be published. JMM strives for scholarship, innovation and excellence. This demands scientific rigour, the best available methods and technologies, correctly replicated experiments/tests, the inclusion of proper controls, calibrations, and the correct statistical analysis. The presentation of the data must support the interpretation of the method/approach. All aspects of microbiology are covered, except virology. These include agricultural microbiology, applied and environmental microbiology, bioassays, bioinformatics, biotechnology, biochemical microbiology, clinical microbiology, diagnostics, food monitoring and quality control microbiology, microbial genetics and genomics, geomicrobiology, microbiome methods regardless of habitat, high through-put sequencing methods and analysis, microbial pathogenesis and host responses, metabolomics, metagenomics, metaproteomics, microbial ecology and diversity, microbial physiology, microbial ultra-structure, microscopic and imaging methods, molecular microbiology, mycology, novel mathematical microbiology and modelling, parasitology, plant-microbe interactions, protein markers/profiles, proteomics, pyrosequencing, public health microbiology, radioisotopes applied to microbiology, robotics applied to microbiological methods,rumen microbiology, microbiological methods for space missions and extreme environments, sampling methods and samplers, soil and sediment microbiology, transcriptomics, veterinary microbiology, sero-diagnostics and typing/identification.