Department of Chemical Engineering and Applied Chemistry at the University of Toronto, 200 College St, Toronto M5S 3E5, Ontario, Canada.
Department of Biology, University of Toronto, Mississauga, 3359 Mississauga Road North, Mississauga L5L 1C6, Ontario, Canada.
Bioresour Technol. 2020 Jan;295:122251. doi: 10.1016/j.biortech.2019.122251. Epub 2019 Oct 9.
Two TaqMan® qPCR assays were developed to specifically quantify the absolute abundance of Scenedesmus obliquus and Chlorella vulgaris in mixed-species algal biofilms by targeting the psbA gene. Standard curves were developed with amplification efficiencies of 92.4% and 96.6% for S. obliquus and C. vulgaris, respectively, and an R value of 0.99 for both. Calibration curves for estimating absolute cell abundances resulted in slopes of 0.98 and 1.11 for C. vulgaris and S. obliquus, respectively, and an R value of 0.95 for both. The assays were applied to cultivated mixed-species biofilms and approximately 10 cells of each algal species were quantified when 10 cells were added into biofilms. The developed qPCR assays were concluded to be specific and accurate for the quantification of S. obliquus and C. vulgaris in mixed-species biofilms. This will contribute to the control and optimization of algal cultivation systems for the production of algal biofuels and bioproducts.
两种 TaqMan® qPCR 检测方法被开发出来，通过靶向 psbA 基因，专门用于定量混合物种藻类生物膜中斜生栅藻和普通小球藻的绝对丰度。针对斜生栅藻和普通小球藻，标准曲线的扩增效率分别为 92.4%和 96.6%，相关系数（R 值）均为 0.99。用于估计绝对细胞丰度的校准曲线的斜率分别为 0.98 和 1.11，相关系数（R 值）均为 0.95。该方法应用于培养的混合物种生物膜中，当向生物膜中添加 10 个细胞时，可定量检测到每种藻类约 10 个细胞。开发的 qPCR 检测方法被证明是专门且准确的，可用于定量混合物种生物膜中的斜生栅藻和普通小球藻。这将有助于控制和优化藻类培养系统，以生产藻类生物燃料和生物制品。
