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Abstract：
Quantifying the growth of entomopathogenic fungi is crucial for understanding their virulence and pathogenic potential. Traditional methods for determining growth, such as biomass determination or colony growth area, are time-consuming and quantitatively and spatially limited in scope. In this study, we introduce a high-throughput method for rapidly measuring fungal growth using spectrophotometry in small-volume, liquid media cultures in 96-well microplates. Optical density (OD) changes were directly correlated with dry weight of samples for six isolates from three species of the genus Metarhizium to validate spectrophotometric growth measurements, and investigate species- and isolate-specific effects. We quantified fungal biomass from the microcultures by extracting, drying, and weighing mycelial mats. From the relationship established between OD and biomass, we generated standard curves for predicting biomass based on the OD values. The OD measurements clearly distinguished growth patterns among six isolates from three Metarhizium species. The logistic growth phase, as captured by the OD measurements, could be accurately assessed within a span of 80 h. Using isolates of M. acridum, M. brunneum, and M. guizhouense, this technique was demonstrated to be an effective, reproducible, and simple method for rapidly measuring filamentous fungal growth with high precision. This technique offers a valuable tool for studying the growth dynamics of entomopathogenic fungi and investigating the factors that influence their growth.
摘要：
定量昆虫病原真菌的生长对于了解其毒力和致病潜力至关重要。确定生长的传统方法，如生物量测定或菌落生长面积，是耗时的，在数量上和空间上都是有限的。在这项研究中,我们介绍了一种高通量方法，使用分光光度法在小体积下快速测量真菌生长，液体培养基培养在96孔微孔板。光密度（OD）的变化与来自三种绿僵菌属的六个分离株的样品干重直接相关，以验证分光光度生长测量。并研究物种和分离株的特定效应。我们通过提取来量化微培养物中的真菌生物量，干燥,和称重菌丝垫。从OD和生物量之间建立的关系来看，我们生成了基于OD值预测生物量的标准曲线。OD测量清楚地区分了来自三种绿僵菌物种的六个分离株之间的生长模式。物流增长阶段，如OD测量所捕获的，可以在80小时的时间内准确评估。使用根草分离株，M.Brunneum,和贵州先生，这项技术被证明是有效的，可重复,方法简便，可快速测量丝状真菌生长，精度高。该技术为研究昆虫病原真菌的生长动力学和研究影响其生长的因素提供了有价值的工具。
