Optimized method for dna extraction and PCR amplification in aroeira tree

Authors

DOI:

https://doi.org/10.14393/BJ-v39n0a2023-62577

Keywords:

CTAB, DNA isolation, Myracrodruon urundeuva.

Abstract

Molecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of β-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers.

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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

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

BHANDARI, S., et al. Role of molecular markers to study genetic diversity in bamboo: a review. Plant Cell Biotechnology and Molecular Biology. 2021, 22(3), 86-97.

CASTRO, C.B., et al. Metabolomics-Based Discovery of Biomarkers with Cytotoxic Potential in Extracts of Myracrodruon urundeuva. Journal of the Brazilian Chemical Society. 2020, 31(4), 775-787. https://doi.org/10.21577/0103-5053.20190242.

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

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.

EGHLIMA, G., et al. Study of genetic diversity of Glycyrrizha glabra L. populations using ISSR molecular markers. Plant Genetic Researches. 2021, 8(1), 81-94.

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

FURTADO FILHO, J.C., et al. Comparação de métodos de isolamento do DNA em Handroanthus serratifolius. Brazilian Journal of Development, 2021, 7(4), 35765-35779. https://doi.org/10.34117/bjdv7n4-168

GOMES, R.L.F., et al. A lima bean core collection based on molecular markers. Scientia Agricola. 2020, 77(2), e20180140. https://doi.org/10.1590/1678-992X-2018-0140

GOMES, M.F.C., et al. Genetic diversity and structure in natural populations of Cajui from Brazilian cerrado. Bioscience Journal. 2021, 37, e37080. https://doi.org/10.14393/BJ-v37n0a2021-53974

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

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. and MATOS, F.J.A. Plantas medicinais no Brasil: nativas e exóticas cultivadas. Nova Odessa: Plantarum. 512p. 2002.

NANODROP TECHNOLOGIES, INC. ND-1000 Spectrophotometer V3.5 User’s Manual. Wilmington, USA, 2007. 61p.

PALMIERI, D.A., et al. Genetic diversity analysis in the section Caulorrhizae (genus Arachis) using microsatellite markers. Genetics and Molecular Biology. 2010, 33(1), 109-118. https://doi.org/10.1590/S1415-47572010005000001

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. and CARNEIRO, V.T.C. 2ed. Manual de transformação genética de plantas. Brasília: EMBRAPA-PI/EMBRAPA - CENARGEN, pp. 163-177. Available from: http://livimagens.sct.embrapa.br/amostras/00053750.pdf

SILVA, C.L.S.P., et al. Avaliação de protocolos para extração de DNA de aroeira (Myracrodruon urundeuva). Cultura Agronômica. 2010, 19(1), 12-22. Available from: https://ojs.unesp.br/index.php/rculturaagronomica/article/view/2163

SILVA, J.N., et al. A simple and cost-effective method for DNA extraction suitable for PCR in sucupira branca. Bioscience Journal. 2021, 37, e37092. https://doi.org/10.14393/BJ-v37n0a2021-54133

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. http://dx.doi.org/10.4238/2014.June.16.7

SOUSA, T.B., et al. Chemical and structural characterization of Myracrodruon urundeuva barks aiming at their potential use and elaboration of a sustainable management plan. Biomass Conversion and Biorefinery. 2022, 12(5), 1583–1593. https://doi.org/10.1007/s13399-020-01093-2

SOUZA, S.M.C., et al. Antiinflammatory and antiulcer properties of tannins from Myracrodruon urundeuva Allemão (Anacardiaceae) in Rodents. Phytotherapy Research. 2007, 21(3), 220-225. https://doi.org/10.1002/ptr.2011

STEFANOVA, P., et al. A modified CTAB method for DNA extraction from soybean and meat products. Biotechnology & Biotechnological Equipment. 2014, 27(3), 3803-3810. https://doi.org/10.5504/BBEQ.2013.0026

TIWARI, K.L., JADHAV, S.K. and GUPTA, S. Modified CTAB Technique for Isolation of DNA from some Medicinal Plants. Research Journal of Medicinal Plants. 2012, 6(1), 65-73. https://doi.org/10.3923/rjmp.2012.65.73

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

VIANA, J.P.G., et al. Divergência genética em germoplasma de alho. Ciência Rural. 2016, 46(2), 203-209. https://doi.org/10.1590/0103-8478cr20130988

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

2023-03-03

How to Cite

SILVA, J. do N., LINHARES, A.C. dos S., PALMIERI, D.A., COSTA, M.F., SÁ, G.H. de, GOMES, M.F. da C., SANTOS, M.F. dos, FEITOZA, L. de L. and DOS SANTOS VALENTE, S.E., 2023. Optimized method for dna extraction and PCR amplification in aroeira tree. Bioscience Journal [online], vol. 39, pp. e39001. [Accessed21 May 2024]. DOI 10.14393/BJ-v39n0a2023-62577. Available from: https://seer.ufu.br/index.php/biosciencejournal/article/view/62577.

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Section

Agricultural Sciences