Vol. 34 No. 2 (2020): Advances in Horticultural Science
Short note

Extraction of total protein from shoots of Cereus morphological variants (Cactaceae) for proteomic analysis: Extraction of protein for proteomic analysis

Maria de Fátima Pires da Silva Machado
Universidade Estadual de Maringá
Dr. V. Neves de Azevedo Fernandes
Postgraduate Program in Genetics and Breeding, Universidade Estadual de Maringá, Maringá PR
Dr. C. Aparecida Mangolin
Department of Biotechnology, Genetics and Cell Biology, Universidade Estadual de Maringá, Maringá PR
Andréa Florindo Neves
Postgraduate Program in Genetics and Breeding, Universidade Estadual de Maringá, Maringá PR
Dr. F.C. Sousa Nogueira
Department of Chemistry of Proteins, Unidad Proteomics, Chemistry Institute, Universidade Federal do Rio de Janeiro, Rio de Janeiro RJ
Dr. Hugo Zeni Neto
Department of Agronomy, Universidade Estadual de Maringá, Maringá PR
Published January 15, 2020
Keywords
  • cactus,
  • phenotypic variants,
  • protein extraction,
  • SDS-PAGE,
  • succulent tissues
How to Cite
Machado, M. de F. P. da S., Neves de Azevedo Fernandes, V., Aparecida Mangolin, C., Florindo Neves, A., Sousa Nogueira, F. C., & Zeni Neto, H. (2020). Extraction of total protein from shoots of Cereus morphological variants (Cactaceae) for proteomic analysis. Advances in Horticultural Science, 34(2), 233-240. https://doi.org/10.13128/ahsc-7848

Abstract

Since there is a hypothesis that qualitative and/or quantitative differences of specific proteins may be associated with morphological variants in cacti of the Cereus genus (phenotypes erect, tortuosus and monstruosus), in current study we tested three different methods for protein extraction from shoots of the phenotypic variants to obtain protein fractions for further proteomic analysis. The TCA/acetone method for protein extraction revealed a larger number of well-defined bands in SDS-PAGE system than the methods with phenol. The quantification of protein extracted by TCA/acetone ranged between 0.488 (tortuosus) and 2.92 µg·mL-1 (monstruosus). Although the use of phenol is the most appropriate procedure for protein extraction from recalcitrant tissues, results have shown that extraction buffer containing two antioxidant agents (EDTA and β-mercaptoethanol) and PMSF to prevent protein degradation was efficient to avoid proteolysis and lower protein yield, than using TCA/acetone precipitation for protein extraction from shoots of Cereus sp. The use of extraction buffer with appropriate combinations of antioxidant agents, phenol-complexing agents, and protease inhibitors may be an efficient alternative for proteins extraction from succulent and recalcitrant tissues (such as cactus plants) using a simple protein extraction method.

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