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Extraction and polyacrylamide gels electrophoresis analysis (SDS-PAGE) of total protein from leaves and roots of Piper tuberculatum

The Pará State is the principal Brazilian producer of black pepper (Piper nigrum Link), however, the spice’s production has been damaged by the Fusarium disease. The Fusarium solani f. sp. piperis, causal agent of this disease which attacks the plant root system promoting the root rot, leaves fall and plant death. Piperaceae plants endemic of Amazon region and resistant to F. solani f. sp. piperis infection, such as Piper tuberculatum Jacq., which has been studied in order to understand this host-pathogen interaction. In this work were evaluated five conditions for total protein extraction aiming to select suitable buffers for extraction of total proteins from P. tuberculatum leaves and roots. The conditions used to roots and leaves protein extractions were salt buffer, sucrose buffer, glycerol buffer, urea buffer and sodium phosphate buffer. Quantitative analysis showed that sucrose, glycerol and urea buffers were more efficient for leaves and roots protein extractions. SDS-PAGE analysis showed distinct band patterns in leaves and roots protein extracts obtained with different buffers. Our results could support the selection of extraction buffers for proteomics analysis in P. tuberculatum - F. solani f sp. piperis interaction studies.

Fusarium disease; Fusarium solani f. sp. piperis; Piper nigrum; Piper tuberculatum; Proteomics; SDS-PAGE


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