Department of Biotechnology and Biophysics, Julius-Maximilian University Würzburg, Biocenter - Am Hubland, D-97074, Würzburg, Germany.
Department of Systemphysiology, Leibniz Institute for Neurobiology, Brenneckestraße 6, D-39118, Magdeburg, Germany.
Sci Rep. 2018 Jan 12;8(1):605. doi: 10.1038/s41598-017-19103-1.
The dynamics of early fungal development and its interference with physiological signals and environmental factors is yet poorly understood. Especially computational analysis tools for the evaluation of the process of early spore germination and germ tube formation are still lacking. For the time-resolved analysis of conidia germination of the filamentous ascomycete Fusarium fujikuroi we developed a straightforward toolbox implemented in ImageJ. It allows for processing of microscopic acquisitions (movies) of conidial germination starting with drift correction and data reduction prior to germling analysis. From the image time series germling related region of interests (ROIs) are extracted, which are analysed for their area, circularity, and timing. ROIs originating from germlings crossing other hyphae or the image boundaries are omitted during analysis. Each conidium/hypha is identified and related to its origin, thus allowing subsequent categorization. The efficiency of HyphaTracker was proofed and the accuracy was tested on simulated germlings at different signal-to-noise ratios. Bright-field microscopic images of conidial germination of rhodopsin-deficient F. fujikuroi mutants and their respective control strains were analysed with HyphaTracker. Consistent with our observation in earlier studies the CarO deficient mutant germinated earlier and grew faster than other, CarO expressing strains.
早期真菌发育的动态及其对生理信号和环境因素的干扰机制还远未被理解。特别是，用于评估早期孢子萌发和芽管形成过程的计算分析工具仍然缺乏。为了对丝状子囊菌藤仓镰刀菌的分生孢子萌发进行时间分辨分析，我们在 ImageJ 中开发了一个简单的工具框。它允许对从漂移校正和数据减少开始的分生孢子萌发的微观采集（电影）进行处理，然后再进行芽体分析。从图像时间序列中提取出与芽体相关的感兴趣区域（ROI），并对其面积、圆度和时间进行分析。在分析过程中，会忽略来自穿过其他菌丝或图像边界的芽体的 ROI。每个分生孢子/菌丝都被识别，并与其来源相关联，从而允许后续进行分类。HyphaTracker 的效率已得到证明，并在不同信噪比下的模拟芽体上进行了准确性测试。用 HyphaTracker 分析了视蛋白缺陷型藤仓镰刀菌突变体及其相应对照菌株的分生孢子萌发的明场显微图像。与我们在早期研究中的观察结果一致，CarO 缺陷型突变体比其他表达 CarO 的菌株更早萌发，生长更快。
