Abstract
Croton urucurana, known as “sangra-d’agua”, is a tree species with timber and beekeeping potential, and also with medicinal potential. The use of microsatellite markers, simple sequence repeats (SSRs) is efficient in population genetic studies, with high transferability rate between correlated species. The present study aimed to optimize the polymerase chain reaction (PCR) protocol and to test the transferability of seven primers developed for Croton floribundus to C. urucurana. The amplification of 113 accessions from different Brazilian biomes (Cerrado, Pantanal and Mata Atlântica) was tested. For optimization, the concentration and label of the MgCl2 reagent was varied in addition to annealing temperature. Four tested primers showed amplification products, guaranteeing 57% of transferability. This result confirms the efficiency of C. floribundus microsatellite loci for future studies of genetic variability in C. urucurana. Key words: Genetic variability, sangra-d’agua, molecular markers, simple sequence repeats (SSRs), primers.
Highlights
Croton urucurana Baill., commonly known by the names "sangra-d'água", "sangre del dragón" or "dragon's blood", is a small tree species from the Euphorbiaceae family, pioneer, found mainly in riparian forests from various forest formations, widely distributed by Paraguay, Argentina, Uruguay and Brazil (Caruzo and Cordeiro, 2007)
Adenohylli, respectively (Van Ee et al, 2011). Considering this and the lack of studies involving the genetic diversity of C. urucurana, the present study aimed to test the transferability of microsatellite markers (SSR) designed for C. floribundus species to C. urucurana, as well as to optimize a polymerase chain reaction (PCR) protocol for the amplification of these primers
Unlike the extract reported in C. floribundus (Silvestrini et al, 2013) and what is expected for pioneer species (Coley, 1993), the DNA extracts obtained in the present study needed to be purified with ammonium acetate for secondary compounds
Summary
Croton urucurana Baill., commonly known by the names "sangra-d'água", "sangre del dragón" or "dragon's blood", is a small tree species from the Euphorbiaceae family, pioneer, found mainly in riparian forests from various forest formations, widely distributed by Paraguay, Argentina, Uruguay and Brazil (Caruzo and Cordeiro, 2007). The species most used are Campylocentrum sellowii (antibiotic activity), Croton tigluim (antitumor), C. cortesianos (treatment of dermatitis), C. macrostahys (vermifuge), C. lineares (antipyretic), C. sublyratus (against ulcer) and C. penduliflorus (purgative) (Randau et al, 2004) Because of this great medicinal importance, genetic studies are essential to develop conservation strategies and products (Alves et al, 2012). Microsatellites or Single Sequence Repeats (SSRs) are very efficient molecular markers in studies of population genetics, genetic variability, evolutionary studies and genetic mapping (Varshney et al, 2005; Hodel et al, 2016) Because of characteristics such as high polymorphism, codominant inheritance pattern, multiple alleles, abundance, and broad distribution throughout the genome, microsatellites are very powerful and popular tools to characterize genetic variation among individuals or closely related taxa (Guichoux et al, 2011). Considering this and the lack of studies involving the genetic diversity of C. urucurana, the present study aimed to test the transferability of microsatellite markers (SSR) designed for C. floribundus species to C. urucurana, as well as to optimize a PCR protocol for the amplification of these primers
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