Extração e análise eletroforética em gel de poliacrilamida (SDS-PAGE) de proteínas totais de folhas e raízes de Piper tuberculatum

Abstract

O Estado do Pará é o principal produtor brasileiro de pimenta-do-reino (Piper nigrum Link), entretanto a sua produção tem sido bastante afetada pela doença conhecida como fusariose. O Fusarium solani f. sp. piperis é o agente causador desta doença que afeta o sistema radicular da planta, causando o apodrecimento das raízes e a queda das folhas levando à morte da planta. Algumas piperáceas nativas da região amazônica, entre elas a espécie Piper tuberculatum Jacq., têm se mostrado resistentes à infecção pelo F. solani f. sp. piperis, e desta forma têm sido utilizadas em estudos de interação planta-patógeno. Neste trabalho foram avaliadas cinco condições de extração de proteínas com o objetivo de selecionar tampões adequados para a extração de proteínas totais de folhas e raízes de P. tuberculatum. Os tampões utilizados para a extração de proteínas de raízes e folhas foram: tampão salino, tampão sacarose, tampão glicerol, tampão uréia e tampão fosfato de sódio. As análises quantitativas mostraram que os tampões sacarose, glicerol e uréia foram mais eficientes na extração de proteínas de folhas e raízes. Análises de SDS-PAGE mostraram padrões diferenciados de bandas em extratos protéicos de folhas e raízes obtidos com os diferentes tampões. Os resultados obtidos neste trabalho contribuem para a identificação de tampões de extração adequados para a obtenção de amostras de proteínas totais em estudos de interação P. tuberculatum -F. solani f. sp. piperis. Extraction and polyacrylamide gels electrophoresis analysis (SDS-PAGE) of total protein from leaves and roots of Piper tuberculatum ABSTRACTThe 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.

New Lectins from Mediterranean Flora. Activity against HT29 Colon Cancer Cells

“…The total protein extract from Arbutus unedo L. were obtained by an optimized method of extraction with buffer containing 5% glycerol and 1% Triton X-100, described by Silva and Souza [36].…”

Influence of co-cultures ofStreptococcus thermophilus and probiotic lactobacilli on quality and antioxidant capacity parameters of lactose-free fermented dairy beverages containingSyzygium cumini(L.) Skeels pulp

“…In the T2 trial, with the presence of L. rhamnosus, it was possible to verify the formation of peptides in the region below 14 kDa on days 14 and 21 of the storage only in the gel stained with silver nitrate, since this staining method has high sensitivity, being able to detect protein fragments of 0.1 to 1.0 ng, whereas Coomassie Brilliant staining detects proteins of 30 to 100 ng. 41 On the other hand, in the T3 trial, with L. casei, proteolytic activity on a-LA was observed in Coomassie Brilliant-stained gel. Moreover, the peptides present in T0, TF and D1 samples of T3 trial have been degraded throughout the storage time and their lower derived fractions could not be detected by silver nitrate staining.…”

By-products of Piper aduncum in the control of fusariosis in black pepper plant

“…The fungus F. solani f. sp. piperis affects the root system of the plant, causing root rot and leaf fall, and ultimate death (Silva & Souza, 2009).…”