Optimization of genomic DNA extraction protocol for black wattle

Authors

DOI:

https://doi.org/10.30612/agrarian.v13i49.9190

Keywords:

Acacia mearnsii. CTAB. Nucleic acids. RNAse.

Abstract

As the integration of classic techniques and molecular genetics analysis is a current trend in forest tree improvement, a protocol for DNA extraction adapted for the study of each species is necessary. The objective of this study was to determine the effect of different adaptations in the CTAB protocol of genomic DNA extraction for black wattle. The following components were tested after the organic extraction phase: chloroform, phenol formaldehyde, and proteinase K. Also, the effects of applying RNase after the precipitation and cleaning phase were analyzed. The efficiency of these treatments was evaluated on fresh leaflets samples, as well as on samples refrigerated for seven days. The authors observed that all treatments enabled the successful genomic DNA extraction using the CTAB protocol. Moreover, storage did not affect the DNA integrity; adding RNase, however, resulted in an enhanced extracted DNA. Therefore, the use of chloroform and RNase according to the CTAB protocol on both fresh and refrigerated samples is suggested.

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Author Biographies

Yasmin Imparato Maximo, Universidade Federal do Paraná (UFPR)

Engenheira Florestal pela UFPR.

Angela Cristina Ikeda, Universidade Federal do Paraná (UFPR)

Doutora em Genética pela UFPR. Técnica do Laboratório de Biotecnologia Florestal da UFPR.

Paulo César Flôres Júnior, Universidade Federal do Paraná (UFPR)

Doutor em Engenharia Florestal pela UFPR.

Giovana Bomfim de Alcantara, Universidade Federal do Paraná (UFPR)

Doutora em Agronomia pela UFPR. Professora do Departamento de Ciências Florestais da UFPR.

Antonio Higa, Universidade Federal do Paraná (UFPR)

Doutor em Engenharia Florestal pela Australian National University. Professor do Programa de Pós-Graduação em Engenharia Florestal da UFPR.

References

AGEFLOR: ASSOCIAÇÃO GAÚCHA DE EMPRESAS FLORESTAIS. A indústria de base florestal no Rio Grande do Sul 2017: ano base 2016. Porto Alegre: AGEFLOR, 2017, 13p.

AUBAKIROVA, K.; OMASHEVA, M.; RYABUSHKINA, N.; TAZHIBAEV, T.; KAMPITOVA, G.; GALIAKPAROV, N. Evaluation of five protocols for DNA extraction from leaves of Malus sieversii, Vitis vinifera, and Armeniaca vulgaris. Genetics and Molecular Research, v.13, n.1, p.1278-1287, 2014.

BORÉM, A.; MIRANDA, G. V. Melhoramento de plantas. 6. ed.. Viçosa: UFV, 2013. 523p.

CAPELOTO, A.; UNÊDA-TREVISOLI, S. H.; ARRIEL, N. H. C.; QUEIROZ, M. A.; DI MAURO, A. O. Quali-quantitative analysis of genomic DNA extraction methods in watermelon. Revista Ceres, v.52, n.299, p.1-12, 2005.

CURY, P. R.; FURUSE, C.; ARAÚJO, N. S. Technique and application of polimerase chain reaction in dentristy. Revista Odontológica de Araçatuba, v.26, n.2, p. 34-39, 2005.

DOYLE, J. J.; DOYLE, J. L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, v.19, p.11-15, 1987.

FLÔRES JR, P. C. Caracterização morfológica e análise de divergência genética entre clones de acácia-negra (Acacia mearnsii De Wildeman). 2015. Ano de obtenção: 2015. 76 f. Dissertação (Mestrado em Engenharia Florestal) – Universidade Federal do Paraná, 2015.

IBÁ: INDÚSTRIA BRASILEIRA DE ÁRVORES. Relatório IBÁ 2016 ano base 2015. Brasília: IBÁ, 2016, 100p.

MAZZA, J. C. M.; BITTENCOURT, J. V. M. Extraction of DNA from plant tissue of Araucaria angustifolia (Araucariaceae). Boletim de Pesquisa Florestal, n.4, p.12-17, 2000.

OLIVEIRA, L. C.; RODRIGUES, D. P.; HOPKINS, M. J. G. Comparison of six DNA extraction protocols to molecular analysis in species of Fabaceae. Scientia Amazonia, v.6, n.3, p.38-45, 2017.

ROMANO, E.; BRASILEIRO, A. C. M. Extração de DNA de plantas. Biotecnologia, v.2, n.9, p.40-43, 1999.

SAHU, S. K.; THANGARAJ, M.; KATHIRESAN, K. DNA Extraction protocol for plants with high levels of secondary metabolites and polysaccharides without using liquid nitrogen and phenol. ISRN Molecular Biology, v.2012, p.1-6, 2012.

SILVA, A. Z. C. da; COSTA, R. B da; CAMPOS, D. T. da S. Comparison of three buffers for extraction of genomic DNA from leaf tissues of Eucalyptus camaldulensis Dehnh. Multitemas, n.48, p.179-193, 2015.

TABERLET, P.; COISSAC, E.; POMPANON, F.; GIELLY, L.; MIQUELL, C.; VALENTINI, A.; VERMAT, T.; CORTHIER, G.;BROCHMANN, C.; WILLERSLEV, E. Power and limitations of the chloroplast trnL (UAA) intron for plant DNA barcoding. Nucleic Acids Research, v.35, p.1-8, 2007.

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Published

2020-07-27

How to Cite

Maximo, Y. I., Ikeda, A. C., Flôres Júnior, P. C., Alcantara, G. B. de, & Higa, A. (2020). Optimization of genomic DNA extraction protocol for black wattle. Agrarian Journal, 13(49), 323–329. https://doi.org/10.30612/agrarian.v13i49.9190

Issue

Vol. 13 No. 49 (2020)

Section

Article - Plant Breeding

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