Quali-quantitative analysis of genomic DNA taken from the leaf and stipe of x Butyagrus nabonnandii (Prosch.) Vorster
DOI:
https://doi.org/10.33448/rsd-v11i10.33005Keywords:
Arecaceae; DNA extraction; Liquid nitrogen; DNA yield; Hybrid palm.Abstract
DNA-based identification tools are of great importance in plant genetic studies, where DNA isolation and purification are crucial steps. The aim of the present study was to test the efficiency of conservation and maceration methods of biological materials in the extraction of genomic DNA from leaf and stipe of x Butyagrus nabonnandii (Prosch.) Vorster. The data were analyzed with the help of the software Genes via variance analysis, considering the factorial scheme 3x2x2x2, with three replications (three specimens of the hybrid; two types of biological material – stipe and leaf; two types of conservation – fresh and dehydrated; two types of maceration – with and without liquid nitrogen). The means were analyzed by Tukey test at 5% error probability. Genomic DNA was evaluated by spectrophotometry to determine the concentration. Integrity was determined by agarose gel eletrophoresis. Two primers from the WRKY gene Family were used to test the quality of the DNA obtained in PCR reactions. Statistical analyses revealed significant differences in DNA concentrations between the evaluated methods. Higher amounts of DNA were obtained from the extraction of fresh leaves when compared to dehydrated ones. DNA yield from fresh and dehydrated stipe did not vary between the methods tested. The use of liquid nitrogen can be dispensed, as a result of band patterns in PCR reactions. It was possible to extract quality DNA and sufficient amounts of biological materials from x B. nabonnandii, which amplified the genomic regions of interest.
References
Aboul-Maaty, N, A. & Oraby, H. A. (2019). Extraction of high-quality genomic DNA from different plant orders applying a modified CTAB-based method. Bulletin of the National Research Centre, 43 (25), 1-10.
Arif, I. A., Bakir, M. A., Khan, H. A., Ahamed, A., Farhan, A. H. A., Homaidan, A. A. A., Sadoon, M. A., Bahkali, A. H. & Shobrak, M. (2010). A simple method for DNA extraction from mature date palm leaves: impact of sand grinding and composition of lysis buffer. International Journal of Molecular Sciences, 11, 3149-3157.
Aydın, Z. U., Şenova, M. K., Koch, M. A. & Dönmez, A. A. (2020). DNA from Leaf or Stem: A Comparative Work on Dianthus L. for DNA Barcoding Analysis with Four Commercial Extraction Kits. Hacettepe Journal of Biology and Chemistry, 48 (4), 333-339.
Azêvedo, H. S. F. S., Rufino, P. B., Azevedo, J. M. A., Silva, L. M. S., WAdt, L. H. O. & Campos, T. (2019). Preservação e maceração de tecido de folíolos de açaí da Amazônia para obtenção de DNA genômico. Bioscience Journal, 35 (4), 1188-1197.
Barbosa Rodrigues, J. (1903). Sertum Palmarum Brasiliensium. Imprimerie Typographique Veuve Monnm, 2 (1),116.
Burret, M. (1940). Um caso de hibridação entre Arecastrum romanzoffianum e Butia capitata. Rodriguesia, 13, 170-173.
Carvalho, M. S., Noia, L. R., Ferreira, M. F. S. & Ferreira, A. (2019). DNA de alta qualidade isolado a partir do córtex de Euterpe edulis Mart. (Arecaceae). Ciência Florestal, 29 (1), 396-402.
Colpaert, S. C., Bandou, E., Caron, H., Gheysen, G. & Lowe, A. J. (2005). Sampling tissue for DNA analysis of trees trunk cambium as an alternative to canopy leaves. Silvae Genetica, 54 (6), 265-269.
Cruz, C. D. (2016). Genes software – extended and integrated with the R, Matlab and Selegen. Acta Scientiarum, 38 (4), 547-552.
Furtado Filho, J. C., Silva, J. N., Viana, J. P. G., Costa, M. F., Sousa, C. C., Gomes, M. F. C, Lopes, A. C. A. & Valente, S. E. S. (2021). Comparação de métodos de isolamento do DNA em Handroanthus Serratifolius. Brazilian Journal of Development, 7 (4), 35765-35779.
Hedge, S., Pai, S. R. & Roy, S. (2017). Combination of DNA isolation and RP-HPLC analysis method for bark samples of Saraca asoca and its adulterant. Biotech, 7 (208), 208.
Ihase, L. O., Horn, R., Anoliefo, G. O, Eke, C. R., Afolab, A. B. & Asemota, O. (2016). Development of a method for DNA extraction from oil palm leaves and the effects of pH and ionic strengh on nucleic acid quantification. Journal of Biological Methods, 3 (2), e37.
Laindorf, B. L., Metz, G. F., Küster, M. C. T., Lucini, F., Freitas, K. E. J., Victória, F. C. & Pereira, A. B. (2019). Transfer of microsatellite markers from other Arecaceae species to Syagrus romanzoffiana (Arecaceae). Genetics and Molecular Research, 18 (3), gmr18183.
Lanes, E. C. M., Nick, C., Kuki, K. N., Freitas, R. D. & Motoike, S. Y. (2013). Genomic DNA isolation of Acrocomia aculeata (Arecaceae) from leaf and stipe tissue samples for PCR analysis. Genetics and Molecular Research, 12 (3), 3905-3911.
Lorenzi, H., Noblick, L. R., Kahn, F. & Ferreira, E. (2010). Flora Brasileira: Arecaceae (Palmeiras). Instituto Plantarum, Nova Odessa, 214-255.
Mangaravite, E., Terra, V., Hattori, E. K. O, Dal’sasso, T. C. S, Bhering, L. L. & Oliveira, L. O. (2020). A feasible method to extract DNA from the cambium of high-canopy trees: from harvest to assessment. Acta Amazonica, 50, 335-338.
Mauro-Herrera, M., Meerow A. W., Borrone, J. W., Kuhn, D. N. & Schnell, R. J. (2006) Ten informative markers developed from WRKY sequences in coconut (Cocos nucifera). Molecular Ecology Notes, 6, 904–906.
Meerow, A. W., Noblick, L., Borrone, J. W., Couvreur, T. L. P., Mauro-Herrera, M., Hahn W. J., Kuhn, D. N., Nakamura, K., Oleas, N. H. & Schnell, R. J. (2009). Phylogenetic analysis of seven WRKY Genes across the palm Subtribe Attaleinae (Arecaceae) identifies Syagrus as sister group of the coconut. Plos ONE, 4 (10), e7353.
Mello, L. M., Reiniger, L. R., Meneghello, G. E., Villela, F. A. & Mota, M. S. (2015). Isolamento de DNA genômico a partir de folhas e câmbio congelados de Erythrina crista-galli L., FABACEAE (Corticeira-do-banhado). Revista Thema, 12 (02), 82-93.
Mo, X. C. & Wangsomnuk, P. P. (2021). A modified non-liquid nitrogen protocol for extraction of high-quality genomic DNA from the inner bark tissues of Dalbergia cochinchinensis (Fabaceae). Genetics and Molecular Research, 20 (2), gmr18836.
Novaes, R. M. L., Rodrigues, J. G. & Lovato, M. B. (2009). An efficient protocol for tissue sampling and DNA isolation from the stem bark of Leguminosae trees. Genetics and Molecular Research, 8 (1), 86-96.
Proschowsky, A. R. (1921). Un beau palmier hybride: Butiarecastrum nabonnandi. Revue Horticole, 93, 290–291.
Soares, K. P., Longhi, S. J., Witeck Neto, L. & Assis, L. C. (2014). Palmeiras (Arecaceae) no Rio Grande do Sul, Brasil. Rodriguésia, 65 (1), 113-139.
Tan, H., Tie, M., Zhang, L., Zhu, Y. & Li, H. (2013). The effects of three diferente grinding methods in DNA extraction of cowpea (Vigna unguiculata L. Walp). African Journal of Biotechnology, 12 (16), 1946-1951.
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Copyright (c) 2022 Patrícia de Oliveira-Neves; Antônio Batista Pereira; Andrés Delgado Cañedo; Velci Queiroz de Souza; Bruna Lucia Laindorf; Maike Brum Azambuja; Lurdes Zanchetta da Rosa
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