A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca”

Authors

  • Jaílson do Nascimento Silva Universidade Federal do Piauí https://orcid.org/0000-0002-6083-5320
  • João Paulo Gomes Viana University of Illinois https://orcid.org/0000-0002-9217-9604
  • Marcones Ferreira Costa Universidade Federal do Piauí https://orcid.org/0000-0003-2953-7330
  • Gisele Holanda de Sá Universidade Federal do Piauí
  • Maria Fernanda da Costa Gomes Universidade Federal do Piauí
  • Lidiane de Lima Feitoza Universidade Federal do Piauí
  • Sérgio Emílio dos Santos Valente Universidade Federal do Piauí

DOI:

https://doi.org/10.14393/BJ-v37n0a2021-54133

Keywords:

Conservation, DNA Isolation, Extraction, ISSR.

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.

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References

ALMEIDA, V.M., et al. Comparison of eight methods to isolate genomic DNA from Hancornia speciosa. Genetics and Molecular Research. 2017, 16(3), 1-7. https://doi.org/10.4238/gmr16039724

ANERAO, J., JHA, V. and DESAI, N. Optimization of DNA extraction methods from Garcinia species for ISSR-PCR, RAPD-PCR and DNA barcoding. Asian Journal of Biotechnology. 2017, 9(1), 35-42. https://doi.org/10.3923/ajbkr.2017.35.42

AZÊVEDO, H.S.F., et al. Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA. Bioscience Journal. 2019, 35(4), 1188-1197. https://doi.org/10.14393/BJ-v35n4a2019-42190

BITENCOURT, J.V.T., et al. Comparison of Different Methodologies for DNA Extraction from Aegla longirostri. Brazilian Archives of Biology and Technology. 2007, 50(6), 989-994. https://doi.org/10.1590/S1516-89132007000700010

BRANDÃO, M. and FERREIRA, P.B.D. Flora apícola do cerrado. Informe Agropecuário, 15(168), 7-14. 1991.

CARVALHO, J.C.T., et al. Anti-inflammatory activity of the crude extract from the fruits of Pterodon emarginatus Vog. Journal of Ethnopharmacology. 1999, 64(2), 127-133. https://doi.org/10.1016/S0378-8741(98)00116-0

CORRÊA, T.C.S. and BERTUCI-JUNIOR, L. Avaliação de quebra de dormência tegumentar em sementes de sucupira branca (Pterodon emarginatus Vog.) submetidas a diferentes tratamentos. Cadernos de Agroecologia. 2012, 7(2), 1-5.

COTA-SÁNCHEZ, J.H., REMARCHUK, K. and UBAYASENA, K. Ready to use DNA extracted with a CTAB method adapted for herbarium specimens and mucilaginous plant tissue. Plant Molecular Biology Reporter. 2006, 24(1), 161-167. https://doi.org/10.1007/BF02914055

DANNER, M.A., et al. Proposta de protocolo para extração de DNA de Jabuticabeira. Ciência Florestal. 2011, 21(2), 363-367.

DELLAPORTA, S.L., WOOD. J. and HICKS, J.B. A plant minipreparation: version II. Plant Molecular Biology Report. 1983, 1(4), 19-20. https://doi.org/10.1007/BF02712670

DONATI, M., et al. Radical scavenging and antimicrobial activities of Croton zehntneri, Pterodon emarginatus and Schinopsis brasiliensis essential oils and their major constituents: estragole, trans-anethole, β-caryophyllene and myrcene. Natural Product Research. 2015, 29(10), 939-946. https://doi.org/10.1080/14786419.2014.964709

DOYLE, J.J. and DOYLE, J.L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin. 1987, 19(1), 11-15.

ELSANHOTY, R.M., RAMADAN, M.F. and JANY, K.D. DNA extraction methods for detecting genetically modified foods: a comparative study. Food Chemitry. 2011, 126(4), 1883-1889. https://doi.org/10.1016/j.foodchem.2010.12.013

FERREIRA, M.E. and GRATTAPAGLIA, D. Introdução ao uso de marcadores RAPD e RFLP em análise genética. 1st ed. Brasília: EMBRAPA-CERNARGEN, 1995.

KHANUJA, S.P.S., et al. Rapid isolation of DNA from dry and fresh samples of plants producing large amounts of secondarity metabolites and essential oils. Plant Molecular Biology Reporter. 1999, 17(1), 1-7. https://doi.org/10.1023/A:1007528101452

LACERDA, C.M.B. and KAGEYAMA, P.Y. Estrutura genética de duas populações naturais de Myracrodruon urundeuva M. Allemão na Região Semi-árida, Brasil. Revista Árvore. 2003, 27(2), 145-150. https://doi.org/10.1590/S0100-67622003000200004

LEZA, A.B., HAJARE, S.T. and CHAUHAN, N.M. Comparative analysis of DNA extraction methods in two popular varieties of finger millet (Eleusine coracana) from Ethiopia. Biotechnology Journal International. 2017, 20(2), 1-7. https://doi.org/10.9734/BJI/2017/33145

LORENZI, H. Árvores brasileiras: manual de identificação e cultivo de plantas arbóreas do Brasil. 2nd ed. Nova Odessa: Instituto Plantarum de Estudos da Flora Ltda, 1992.

MAFRA, I., et al. Comparative study of DNA extraction methods for soybean derived food products. Food Control. 2008, 19(12), 1183-1190. https://doi.org/10.1016/j.foodcont.2008.01.004

MONDRAGON-JACOBO, C., DOUDAREVA, N. and BORDELON, B.P. DNA extraction from several cacti. HortScience. 2000, 35(6), 1124-1126. https://doi.org/10.21273/HORTSCI.35.6.1124

OLIVEIRA, A.E., et al. Development of a larvicidal nanoemulsion with Pterodon emarginatus Vogel Oil. PLoS One. 2016, 11(1), 1-16. https://doi.org/10.1371/journal.pone.0145835

OLIVEIRA, G.R.D., et al. Dietary supplementation with plant oils from copaíba (Copaifera langsdorffii) and sucupira (Pterodon emarginatus): quality of physical aspects of eggs stored under different temperatures. Ciência Animal Brasileira. 2018, 19(1), 1-12. https://doi.org/10.1590/1809-6891v19e-41508

RAIMUNDO, J., REIS, C.M.G. and RIBEIRO, M.M. Rapid, simple and potentially universal method for DNA extraction from Opuntia spp. fresh cladode tissues suitable for PCR amplification. Molecular Biology Reports. 2018, 45(5), 1405-1412. https://doi.org/10.1007/s11033-018-4303-8

ROMANO, E. and BRASILEIRO, A.C., 2015. Extração de DNA de tecidos vegetais. In: BRASILEIRO, A.C.M., CARNEIRO, V.T.C. (Eds.). Manual de transformação genética de plantas. 2nd ed. Brasília: EMBRAPA-PI/EMBRAPA – CENARGEN, pp. 163 -177.

SHEDBALKAR, U.U., et al. Opuntia and other cacti: applications and biotechnological insights. Tropical Plant Biology. 2010, 3(1), 136-150. https://doi.org/10.1007/s12042-010-9055-0

SILVA, I.D., et al. Efeito do extrato de sucupira (Pterodon emarginatus Vog.) sobre o desenvolvimento de fungos e bactérias fitopatogênicos. Pesquisa Agropecuária Tropical. 2005, 35(2), 109-115.

SILVA-JUNIOR, M.C. 100 Árvores do cerrado: guia de campo. 1st ed. Brasília: LGE, 2005.

STEFANOVA, P., et al. A modified CTAB method for DNA extraction from soybean and meat products. Biotechnology & Biotechnological Equipment. 2013, 27(3), 3803-3810. https://doi.org/10.5504/BBEQ.2013.0026

SOUSA, C.C., et al. Short Communication Comparison of methods to isolate DNA from Caesalpinia ferrea. Genetics and Molecular Research. 2014, 13(2), 4486-4493. https://doi.org/10.4238/2014.june.16.7

SYAMKUMAR, S., LOWARENCE, B. and SASIKUMAR, B. Isolation and amplification of DNA from rhizomes of turmeric and ginger. Plant Molecular Biology Reporter. 2003, 21(1), 171a–171e. https://doi.org/10.1007/BF02774243

VIANA, J.P.G., et al. Comparison of eight methods of genomic DNA extraction from babassu. Genetics and Molecular Research. 2015, 14(4), 18003-18008. https://doi.org/10.4238/2015.december.22.26

XIA, Y., et al. A modified SDS-based DNA extraction method from raw soybean. Bioscience Reports. 2019, 39(2), 1-10. https://doi.org/10.1042/BSR20182271

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Published

2021-12-29

How to Cite

DO NASCIMENTO SILVA, J.., GOMES VIANA, J.P., COSTA , M.F.., SÁ, G.H. de ., GOMES, M.F. da C.., FEITOZA , L. de L.. and VALENTE , S.E. dos S.., 2021. A simple and cost-effective method for DNA extraction suitable for PCR in “sucupira branca”. Bioscience Journal [online], vol. 37, pp. e37092. [Accessed8 August 2022]. DOI 10.14393/BJ-v37n0a2021-54133. Available from: https://seer.ufu.br/index.php/biosciencejournal/article/view/54133.

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Section

Agricultural Sciences