Artículos Científicos

Permanent URI for this collectionhttp://20.38.43.173:4000/handle/UCP/879

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A simple and efficient method for high-quality total RNA isolation from oleaginous microalgae
(Plant Cell Biotechnology and Molecular Biology, 18(1-2), 15-21, 2017-01-06) Castro, Juan C.; Rodríguez, Hicler N.; Maddox, J. Dylan; Jiu, Bruce; Petterman, Joe B.; Marapara, Jorge L.; Cobos, Marianela
Microalgae are a very diverse group of microorganisms with great potential for biotechnological applications. To best exploit these organisms is necessary to expand our knowledge at its molecular levels. To realize these studies, however, it is essential to have efficient and reproducible methods for total RNA isolation. The objective of this research was to develop a method for high-quality total RNA isolation from three oleaginous microalgae species promissories for biotechnological applications. Three strains Ankistrodesmus sp., Chlorella sp., and Scenedesmus sp. were cultured on CHU-10 medium. Then, were harvested by centrifugation and total RNA was isolated with three commercial kits, one previously published method, and our improved CTAB-LiCl method. Quality and quantity of total RNA isolated were assayed with standard spectrophotometric and electrophoretic techniques. The highest quality (A260/A280: 1.95-2.02, A260/A230: 1.12-1.84) and yield (16.40-93.63 mg/g fresh weight) of total RNA from the three microalgae strains were obtained with our improved CTAB-LiCl method. In addition, the time required to complete the isolation process and the cost were relatively lowest. In conclusion, the improved CTAB-LiCl method, in contrast to frequently used commercial kits, and a previously published method, is effective and reliable to produce total RNA of high quality and quantity that is suitable for cDNA synthesis and polymerase chain reaction.
De novo RNA-Seq analysis of the oleauginous microalgae Ankistrodesmus sp. UCP0001: Gene identification and metabolic pathways reconstruction for the biosynthesis of fatty acids and triacylglycerols
(Plant Cell Biotechnology and Molecular Biology 18(5-6): 219-230, 2017-05-15) Castro, Juan C.; Maddox, J. Dylan; Paredes, Jae D.; Rodríguez, Hicler N.; Aguilar, Carla P.; Marapara, Jorge L.; Castro, Carlos G.; Casuso, María Z.; Cobos, Marianela
Microalgae have great potential as feedstock to produce next-generation biofuels. The scarceness of genomic level information, however, prevents the rational de novo microalgae strain design. In this research, we describe the next-generation sequencing, de novo assembly, and functional annotation of the transcriptome of Ankistrodesmus sp. UCP0001. In total 48,867,830 high-quality sequence reads were de novo assembled into 38,414 unigenes (mean length = 508 bp, N50 = 1038 bp). Seventy-two percent of unigenes presented mapping information. Based on the KEGG pathway assignment, the fatty acids and the triacylglycerol biosynthesis pathways were reconstructed. Our results demonstrate that the synergy among high-throughput sequencing technologies and appropriate bioinformatics tools provides a fast, low-cost, and effective approach to generate invaluable functional genomic information in non-model microalgae species (e.g., Ankistrodesmus sp.). With the de novo assembled and annotated transcriptome we have successfully identified genes encoding enzymes and reconstructed metabolic pathways for the biosynthesis of fatty acids and triacylglycerols in this microalgae species. This genetic information could be used for the de novo microalgae strain design with desirable characteristics to produce biodiesel and capabilities for the biosynthesis of others valuable bioactive compounds of interest to the pharmacological, food, and cosmetic industries.