Vongsvivut Jitraporn, Heraud Philip, Gupta Adarsha, Thyagarajan Tamilselvi, Puri Munish, McNaughton Don, Barrow Colin J
Centre for Chemistry and Biotechnology (CCB), School of Life and Environmental Sciences, Deakin University, Pigdons Road, Waurn Ponds, Victoria 3216, Australia.
Centre for Biospectroscopy, School of Chemistry, Monash University, Wellington Road, Clayton, Victoria 3800, Australia.
Protist. 2015 Feb;166(1):106-21. doi: 10.1016/j.protis.2014.12.002. Epub 2014 Dec 11.
The superior characteristics of high photon flux and diffraction-limited spatial resolution achieved by synchrotron-FTIR microspectroscopy allowed molecular characterization of individual live thraustochytrids. Principal component analysis revealed distinct separation of the single live cell spectra into their corresponding strains, comprised of new Australasian thraustochytrids (AMCQS5-5 and S7) and standard cultures (AH-2 and S31). Unsupervised hierarchical cluster analysis (UHCA) indicated close similarities between S7 and AH-7 strains, with AMCQS5-5 being distinctly different. UHCA correlation conformed well to the fatty acid profiles, indicating the type of fatty acids as a critical factor in chemotaxonomic discrimination of these thraustochytrids and also revealing the distinctively high polyunsaturated fatty acid content as key identity of AMCQS5-5. Partial least squares discriminant analysis using cross-validation approach between two replicate datasets was demonstrated to be a powerful classification method leading to models of high robustness and 100% predictive accuracy for strain identification. The results emphasized the exceptional S-FTIR capability to perform real-time in vivo measurement of single live cells directly within their original medium, providing unique information on cell variability among the population of each isolate and evidence of spontaneous lipid peroxidation that could lead to deeper understanding of lipid production and oxidation in thraustochytrids for single-cell oil development.
同步加速器傅里叶变换红外光谱（synchrotron-FTIR）实现的高光子通量和衍射极限空间分辨率的卓越特性，使得对单个活的破囊壶菌进行分子表征成为可能。主成分分析显示，单个活细胞光谱能够根据相应菌株明显区分开来，这些菌株包括新的澳大利亚破囊壶菌（AMCQS5-5和S7）以及标准培养物（AH-2和S31）。无监督层次聚类分析（UHCA）表明，S7和AH-7菌株之间相似度很高，而AMCQS5-5则明显不同。UHCA相关性与脂肪酸谱吻合良好，表明脂肪酸类型是这些破囊壶菌化学分类鉴别中的关键因素，同时也揭示出AMCQS5-5的显著特征是高含量的多不饱和脂肪酸。使用交叉验证方法在两个重复数据集之间进行的偏最小二乘判别分析被证明是一种强大的分类方法，能够生成稳健性高且菌株鉴定预测准确率达100%的模型。结果强调了同步加速器傅里叶变换红外光谱具有卓越能力，可在单个活细胞的原始培养基中直接进行实时体内测量，提供关于每个分离株群体中细胞变异性的独特信息，以及自发脂质过氧化的证据，这有助于更深入地理解破囊壶菌中脂质产生和氧化过程，以用于单细胞油的开发。
