Selection of protein extraction method for electrophoretic analysis of seeds of the Physic nut genus
Abstract
Abstract: The study aimed to partially extract and purify the proteins present in the seed of Jatropha curcas L., select the best extraction method to characterize the protein electrophoretic profile of genotypes belonging to the Center for Genetic Improvement and Biotechnology, prioritizing ribosome-inactivating proteins (RIPs). The standardization was conducted in four ways, which distinguished in the form of protein extraction and in the extraction solutions, which are: I) Sodium Acetate Buffer (0.45 M) and II), HCl (0.1%)/NaCl (0.6 M), III) NaOH (1 M) and IV) Phosphate-saline buffer (PBS). The method of protein extraction with the best electrophoretic resolution, which was with PBS, using albumen from the stored seeds. The protein concentrations of the samples ranged from 2.75 to 5.15 mg / mL between the genotypes but did not differ statistically from each other by the Tukey test at 5% probability. It was possible to identify RIPs through the electrophoretic profile, with a molecular weight of 28 kDa, in all evaluated genotypes. Profiles from different bands were observed in the UFRB05 and UFRB15 genotypes. The UFRB05 genotype stood out when associated with the quality of stored seeds and oil content.
Downloads
References
ALMEIDA, A.Q.; SILVA, S.A.; ALMEIDA, V.O.; SOUZA, D.R.; ARAÚJO, G.M. Genetic divergence and morpho-agronomic performance of Jatropha curcas L. clones for selection of clonal varieties. Revista Caatinga, v. 29, p. 841-849, 2016. DOI: 10.1590/1983-21252016v29n408rc.
CAVALCANTI, M.T.; BORA, P.S. Análise das proteínas e estudo reológico dos isolados proteicos das amêndoas da faveleira (Cnidosculus phyllacanthus (Mart.) Pax. et K. Hoffm.) com e sem espinhos. Revista do Instituto Adolfo Lutz (Impresso), v. 69, p. 243-51, 2010. DOI: 10.53393/rial.2010.v69.32663.
FARIAS, L.F. Biologia floral, reprodutiva, visitantes florais e desempenho morfoagrônico de Jatropha curcas L. (Euphorbiaceae). 2018. 94 f. Dissertação (Mestrado em Ciências Agrárias) - Universidade Federal do Recôncavo da Bahia, Cruz das Almas, 2018.
LAEMMLI U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, v. 227, p. 680-5, 1970. DOI: 10.1038/227680a0.
LAVIOLA, B.G.; RODRIGUES, E.V.; TEODORO, P.E.; PEIXOTO, L.A.; BHERING, L.L. Biometric and biotechnology strategies in Jatropha genetic breeding for biodiesel production. Renewable and Sustainable Energy Reviews, v. 76, p. 894-904, 2017. DOI: 10.1016/j.rser.2017.03.116.
LAVIOLA, B.G.; SILVA, D.A.S.; JUHASZ, A.C.P.; ROCHA, R.B.; OLIVEIRA, R.J.P.; ALBRECHT, J.C.; ALVES, A.A.; ROSADO, T.B. Desempenho agronômico e ganho genético pela seleção de pinhão-manso em três regiões do Brasil. Pesquisa Agropecuária Brasileira, v. 49, p. 356-363, 2014. DOI: 0.1590/S0100-204X2014000500005.
LIN, J.; ZHOU, X.; WANG, J.; JIANG, P.; TANG, K. Purification and characterization of curcin, a toxic lectin from the seed of Jatropha curcas. Preparative Biochemistry and Biotechnology, v. 40, p. 107–18, 2010. DOI: 10.1080/10826060903558588.
LIU, W.Y.Research on ribosome-inactivating proteins from angiospermae to gymnospermae and cryptogamia. American Journal of Translational Research, v. 9, p. 5719-5742, 2017. Disponível em: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5752922/. Acesso em: 04 out. 2020.
OLATUNJI, T.;MORAKINYO, J. Crude Protein Profiling of Varieties of Capsicum annuum and Capsicum frutescens using SDS-PAGE. IOSR Journal of Pharmacy and Biological Sciences, v. 10, p. 64-71, 2015. DOI: 10.9790/3008-10316471.
PEREIRA, I.R.; D’ABADIA, P.L.; DO PRADO, A.D.L.; MATOS, F.S.; NABOUT, J.C.; GONÇALVES, P.J.; ALMEIDA, L. M. Trends and gaps in the global scientific literature about Jatropha curcas L.(Euphorbiaceae), a tropical plant of economic importance. Semina: Ciências Agrárias, v. 39, n. 1, p. 7-17, 2018. DOI: 10.5433/1679-0359.2018v39n1p7.
PESTANA-CALDAS, C.N.; SILVA, S.A.; MACHADO, E.L; DE SOUZA, D.R; CERQUEIRA-PEREIRA, E.C.; SILVA, M.S. Geneticdivergencethrough joint analysisofmorphoagronomicand molecular characters in accessionsofJatropha curcas. Genetics and Molecular Research, v. 4, p. 1-11, 2016. DOI: 10.4238/gmr.15048385.
R Development Core Team. R: A Language and Environment for statistical Computing. R foundation for Statistical Computing, Vienna: R Foundation, 2020.
SOUZA, L.M.; CARVALHO, L.P.; ARAÚJO, J.S.; MELO, E.J.T.; MACHADO, O.L.T. Cell toxicity by ricin and elucidation of mechanism of Ricin inactivation. International journal of biological macromolecules, v. 113, p. 821-828, 2018. DOI: 10.1016/j.ijbiomac.2018.03.024.
RUST, A.; PARTRIDGE, L.J.; DAVLETOV, B.; HAUTBERGUE, G.M. The use of plant-derived ribosome inactivating proteins in immunotoxin development: Past, present and future generations. Toxins, v. 9, n. 11, p. 344, 2017. DOI: 10.3390/toxins9110344.
STEPHAN, M.P.; SILVA, B.M.; AZEVEDO, T.L.; ASCHERI, J.L.M. Metodologia de extração de proteínas em torta de mamona e pinhão manso para análise por eletroforese (SDS-PAGE). 2010. Disponível em: https://www.infoteca.cnptia.embrapa.br/bitstream/doc/874481/1/pub151.pdf. Acesso em: 06 mar. 2020.
SUHARTI, T.; DJAM'AN, D.D. Potensi RIP (ribosome inactivating protein) yang berasaldaritumbuhansebagaibiopestisida (The Potential of RIP (Ribosome Inactivating Protein) as Biopesticides). BuletinEboni, v.1, p. 33-39, 2019. DOI: 10.20886/buleboni.
VIRGENS, I.O.; CASTRO, R.D.D.; LOUREIRO, M.B.; FERNANDEZ, L.G. Revisão: Jatropha curcas L.: aspectos morfofisiológicos e químicos. Brazilian Journal of Food Technology, v. 20, p. 1-11, 2017. DOI: 10.1590/1981-6723.3016.
WANI, T.A.; KITCHLU, S.; RAM, G. Genetic variability studies for morphological and qualitative attributes among Jatropha curcas L. accessions grown under subtropical conditions of North India. South African Journal of Botany, v. 79, p. 102-105, 2012. DOI: 10.1016/j.sajb.2011.10.009.
WANG, X.H.; LIU, J.Q.; CHEN, S.; YIN, Y.; LIU, Y.; ZHANG, C. Hydroxy-octadecenoic acids instead of phorbol esters are responsible for the Jatropha curcas kernel cake’s toxicity. Communications biology, v. 3, n. 1, p. 1-14, 2020. DOI: 10.1038/s42003-020-0919-z.
ZHU, F.; ZHOU, Y.K.; JI, Z.L.; CHEN X.R. The Plant Ribosome-Inactivating Proteins Play Important Roles in Defense against Pathogens and Insect Pest Attacks. Frontiers in Plant Science, v. 9, p. 146, 2018. DOI: 10.3389/fpls.2018.00146.
ZHU, F.; ZHU, P.X.; XU, F.; CHE, Y.P.; MA, Y.M.; JI, Z.L. Alpha momorcharin enhances Nicotiana benthamiana resistance to tobacco mosaic virus infection through modulation of reactive oxygen species. Molecular Plant Pathology, v. 21, p. 1212-1226, 2020. DOI: 10.1111/mpp.12974.
Copyright (c) 2023 Francielly Carvalho Oliveira, Simone Alves Silva, Ciro Ribeiro Filadelfo, Jacqueline Araújo Castro, Cecília Dominical Poy, Hilçana Ylka Gonçalves de Albuquerque
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors retain copyright and grant the Journal the right to the first publication. Authors are encouraged to and may self-archive a created version of their article in their institutional repository, or as a book chapter, as long as acknowledgement is given to the original source of publication. As the Journal provides open access to its publications, articles may not be used for commercial purposes. The contents published are the sole and exclusive responsibility of their authors; however, the publishers can make textual adjustments, adaptation to publishing standards and adjustments of spelling and grammar, to maintain the standard patterns of the language and the journal. Failure to comply with this commitment will submit the offenders to sanctions and penalties under the Brazilian legislation (Law of Copyright Protection; nº 9,610; 19 February 1998).
.jpg)
