High-resolution melting analysis for identification of microalgae species
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High-resolution melting analysis for identification of microalgae species Daniela Rojo 1 & Manuel Zapata 1 & Alejandro Maureira 1 & Ricardo Guiñez 2 & Cristian Wulff-Zottele 3 & Mariella Rivas 1 Received: 30 December 2019 / Revised and accepted: 18 August 2020 # Springer Nature B.V. 2020
Abstract There is great microalgae biodiversity and their rediscovery as an inexhaustible source of biotechnological resources for all types of applications is leading to the already demonstrated benefits of their bioactive compounds which has boosted their industrial development. For this development, it is essential to have available technologies to easily control large-scale cultures. In this study, a high-resolution melting (HRM) analysis was developed to identify different microalgae that are currently used by the industry. HRM analysis is a simple, economical, fast, and reproducible method that allows the use of a stable molecule such as DNA to track a culture day by day without the need for visual microscopic controls. Ten microalgae were characterized by qPCRHRM analysis: Isochrysis galbana, Phaeodactylum tricornutum, Muriellopsis sp., Porphyridium cruentum, Botryococcus braunii, Nannochloropsis gaditana, Chlorella sorokiniana, Chlamydomonas reinhardtii, Haematococcus pluvialis, and Scenedesmus obliquus. We determined that through the use of primers designed specifically for the 18S rDNA ribosomal gene, it is possible to significantly discriminate among the ten strains simultaneously by using HRM analysis. The results are also replicable over time, facilitating the daily and low-cost control of species used in the biotechnology industry. Keywords High-resolution melting . Microalgae identification . 18S rDNA ribosomal gene . Real-time PCR
Introduction Microalgae are unicellular organisms adapted to live in aquatic systems, on land, or as symbionts (Centella et al. 2017). They perform photosynthesis which produces oxygen and consumes CO2 and their cultivation produces biomass that contains highvalue bioactive compounds such as carotenoid pigments (Guedes et al. 2011; Christaki et al. 2013), lipids, ω3 and ω6 Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10811-020-02240-y) contains supplementary material, which is available to authorized users. * Mariella Rivas [email protected] 1
Laboratorio de Biotecnología Ambiental Aplicada, Departamento de Biotecnología, Facultad de Ciencias del Mar y Recursos Biológicos, Universidad de Antofagasta, Avda. Angamos, 601 Antofagasta, Chile
2
Instituto de Ciencias Naturales Alexander von Humbolt, Facultad de Ciencias del Mar y Recursos Biológicos, Universidad de Antofagasta, Antofagasta, Chile
3
Departmento de Ciencias Biomédicas, Facultad de la Salud, Universidad de Antofagasta, Avda. Angamos, 601 Antofagasta, Chile
polyunsaturated fatty acids (PUFA) (Sharma et al. 2012), proteins (Becker 2004) and carbohydrates, with numerous properties that are applicable and of great interest to the pharmaceutical, food and agricultural indu
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