Standardization of a protein extraction protocol for the proteomic analysis of the leaves of Oryza sativa L. cv. BRS AG. Issue 8 (5th July 2021)

• Record Type: Journal Article
• Authors: Vighi, Isabel Lopes; Seixas Neto, Amilton; de Sousa, Guilherme Feijó; de Carvalho, Juliana Oliveira; da Silva Pinto, Luciano
• Is Part Of: Separation science plus. Volume 4:Issue 8(2021)
• Page Start: 305
• Page End: 313
• Publication Date: 2021-07-05
• Subjects: protein extraction -- proteomics -- rice -- two‐dimensional electrophoresis; Chromatographic analysis -- Periodicals; 543.8
• DOI: 10.1002/sscp.202100019
• Languages: English
• ISSNs: 2573-1815

Abstract

Abstract: The rice cultivar BRS AG ( Oryza sativa L.) is a cereal developed to serve as a raw material for ethanol production or animal feed, contributing to an increasing energy demand and the economic sustainability of the rice sector. However, only few studies have reported this new cultivar. As proteomic analysis is crucial in understanding the physiological and adaptive behavior of BRS AG rice leaves under biotic and abiotic stresses, the present study compared the following three precipitation protocols for the protein extraction of leaf tissue in two‐dimensional electrophoresis analysis: phenol method, trichloroacetic acid‐acetone method, and a third method developed in our laboratory (BIOPRO). The efficiency and repeatability of the protein extraction protocols were evaluated by two‐dimensional electrophoresis analysis. Our results indicate that the BIOPRO protocol was the most efficient in protein extraction as compared to the efficiency of other protocols. We detected a greater amount of protein (15.44 μg/μL) and 414 spots, and the BIOPRO protocol provided the best results regarding reproducibility, focusing, resolved spots, the intensity of spots, and single spot detection. This novel protocol is useful for extracting high‐quality proteins from leaves and presents an efficient removal of contaminants, which is directly linked to a better resolution for two‐dimensional electrophoresis analysis.