Department of Food Science, Stellenbosch University, Private Bag X1, Matieland (Stellenbosch), 7602, South Africa.
Appl Microbiol Biotechnol. 2012 Nov;96(3):803-13. doi: 10.1007/s00253-012-4380-x. Epub 2012 Sep 8.
Colony growth of three Fusarium spp. on potato dextrose agar was followed by collecting near-infrared (NIR) hyperspectral images of the colonies at regular intervals after inoculation up to 55 h. After principal component analysis (PCA), two clusters were apparent in the score plot along principal component 1. Using the brushing technique, these clusters were divided into four groups of pixels with similar score values. These could be visualised as growth zones within the colonies in the corresponding score image. Three spectral bands, i.e. 1,166, 1,380 and 1,918 nm, were prominent in the multiplicative scatter corrected and Savitzky-Golay second derivative spectra. These indicated chemical changes, associated with carbohydrates (1,166 and 1,380 nm) and protein (1,918 nm), that occurred as the mycelium grew and matured. The protein band was more prominent in the mature fungal material while the carbohydrate band was less pronounced. The younger material and the agar were characterised by the carbohydrate spectral band. Integrating whole mycelium colonies as the sum of pixels over time made it possible to construct curves that resembled growth curves; this included the lag phase, active growth phase, deceleration phase and phase of constant growth. Growth profiles constructed from individual growth zones indicated more detailed growth characteristics. The use of NIR hyperspectral imaging and multivariate image analysis (MIA) allowed one to visualise radial growth rings in the PCA score images. This would not have been possible with bulk spectroscopy. Interpreting spectral data enabled better understanding of microbial growth characteristics on agar medium. NIR hyperspectral imaging combined with MIA is a powerful tool for the evaluation of growth characteristics of fungi.
三种镰刀菌在马铃薯葡萄糖琼脂上的菌落生长情况,在接种后每隔一段时间采集近红外(NIR)高光谱图像,直到 55 小时。主成分分析(PCA)后,在沿主成分 1 的得分图上,两个聚类在得分图上明显。使用刷涂技术,这些聚类可以分为具有相似得分值的四个像素组。这些可以在相应的得分图像中可视化为菌落内的生长区。三个光谱带,即 1,166、1,380 和 1,918nm,在乘法散射校正和 Savitzky-Golay 二阶导数光谱中很突出。这些表明了与碳水化合物(1,166 和 1,380nm)和蛋白质(1,918nm)相关的化学变化,这些变化发生在菌丝生长和成熟过程中。蛋白质带在成熟真菌材料中更为突出,而碳水化合物带则不那么明显。较年轻的材料和琼脂由碳水化合物光谱带表征。随着时间的推移,将整个菌丝体菌落作为像素总和进行积分,就可以构建类似于生长曲线的曲线;这包括迟滞期、活跃生长期、减速期和恒速生长期。从单个生长区构建的生长曲线表明了更详细的生长特征。使用近红外高光谱成像和多元图像分析(MIA)可以在 PCA 得分图像中可视化径向生长环。这是批量光谱学无法做到的。解释光谱数据可以更好地了解微生物在琼脂培养基上的生长特性。近红外高光谱成像与 MIA 相结合,是评估真菌生长特性的有力工具。