Abstract
“Sucupira branca” is a plant found in the Brazilian Cerrado and is adapted to low fertility soils, and its fruit extract has anti-inflammatory, healing, antiulcerogenic, antimicrobial, cercaricidal, leishmanicidal and antioxidant activities. Furthermore, it provides protection against oxidative stress, is a natural biocontrol agent of Aedes aegypti, has very resistant wood, is a melliferous plant and has been used in reforestation programs. The development of conservation strategies is important for maintaining diversity in natural populations of “sucupira branca” since these populations are in the process of genetic erosion. Molecular biology techniques, which are important for characterizing the genetic diversity of plants to develop conservation strategies, require sufficient high-quality genomic deoxyribonucleic acid (DNA). This study aimed to compare five methods to extract DNA from “sucupira branca”. The quality and concentration of DNA were revealed by agarose gel electrophoresis, and only the protocols of Dellaporta, Wood and Hicks et al. (1983) and Khanuja et al. (1999) did not result in satisfactory quantities of DNA. When PCR (Polymerase Chain Reaction) was performed with three inter-simple sequence repeat (ISSR) primers, DNA was successfully amplified from extractions performed with the protocols proposed by Doyle and Doyle (1987), Romano and Brasileiro (1998) and Ferreira and Grattapaglia (1995), which are less expensive than commercial purification kits. These protocols resulted in DNA of sufficient quality and quantity after the amplification reactions were performed.
Highlights
Pterodon emarginatus Vog. (Leguminosae) is a tree popularly known as “sucupira branca”
When polymerase chain reaction (PCR) (Polymerase Chain Reaction) was performed with three inter-simple sequence repeat (ISSR) primers, deoxyribonucleic acid (DNA) was successfully amplified from extractions performed with the protocols proposed by Doyle and Doyle (1987), Romano and Brasileiro (1998) and Ferreira and Grattapaglia (1995), which are less expensive than commercial purification kits
Because no research has focused on selecting a DNA extraction method for “sucupira branca”, this work aimed to compare the efficiency of five protocols for DNA extraction by PCR amplification in “sucupira branca” leaves using three inter-simple sequence repeat (ISSR) primers, aiming to achieve an efficient, low-cost procedure to obtain high-quality DNA for molecular analysis
Summary
Pterodon emarginatus Vog. (Leguminosae) is a tree popularly known as “sucupira branca”. (Leguminosae) is a tree popularly known as “sucupira branca”. It is widely found in the Brazilian Cerrado and is adapted to low fertility soils (Brandão and Ferreira 1991). “Sucupira branca” is widely used in traditional medicine in Brazil since its fruit extract has antiinflammatory, healing, antiulcerogenic, antimicrobial, cercaricidal, and antioxidant activities (Carvalho et al 1999; Donati et al 2015; Oliveira et al 2018). A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca”. It is a melliferous plant with potential for urban reforestation and landscaping; it has very resistant and long-lasting wood, which is used in the manufacture of furniture and in civil and naval construction (Silva-Junior 2005). Because it is a highly resistant plant tolerant to direct light, “sucupira branca” has been used in reforestation programs in degraded areas and permanent preservation areas (Lorenzi 1992)
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