New primers tests for the rbcL gene amplification in Chlorophyta

• Grant number: 15/19263-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2016
• End date: December 31, 2016
• Field of knowledge: Biological Sciences - Genetics - Plant Genetics
• Principal Investigator: Armando Augusto Henriques Vieira
• Grantee: Nathan Eugeni Andolfato
• Host Institution: Centro de Ciências Biológicas e da Saúde (CCBS). Universidade Federal de São Carlos (UFSCAR). São Carlos , SP, Brazil
• Associated research grant: 11/50054-4 - Biodiversity of freshwater microalgae: germplasm bank and obtaining of molecular markers of cryopreserved species, AP.BTA.TEM

Abstract

The rbcL gene has been widely used for phylogeny of green algae, and is one of the potential markers for DNA barcoding these algae. However, for the complete amplification of the rbcL many studies using various combinations of primers, and even gene fragments, failures in the amplification may occur due to the presence of introns. From rbcL sequences obtained from Genbank and sequences obtained in other works inside the thematic, degenerate primers were designed in order to increase coverage / universality of markers and to avoid regions with introns (regions to be tested as DNA barcoding) or to facilitate amplification of introns and, consequently, obtaining the complete gene (for phylogenetic analysis). The objective of this project is to test 7 new primers, and a pair of primers already described in the literature, in microalgae of 3 classes of Chlorophyta. Also, after the choice of primers with greater universality within the group, we intend to test them in other microalgae of Selenastraceae family (Chlorophyceae, Chlorophyta) to obtain sequences that will assist us phylogeny projects and DNA barcoding already underway on the project thematic. (AU)