Two‐dimensional electrophoresis of acidic proteins isolated from ozone‐stressed Norway spruce needles (Picea abies L. Karst): Separation method and image processing

Abstract

The effect of ozone stress (< 10,200 or 400 micrograms m-3) on the protein pattern of Picea abies L. Karst needles was studied by fumigating potted grafts from mature trees. Clonal trees of Norway spruce were fumigated in open-top chambers for one growing season. Proteins, soluble and membrane bound, were extracted from the needles in a four-step procedure: (i) grinding in liquid nitrogen and then in a pH 5.0 buffer with thiourea added to inhibit phenol oxidase and polyvinylpolypyrrolidone (PVP) added to bind phenolic compounds; (ii) precipitation of proteins with acetone at -20 degrees C; (iii) solubilization in a modified lysis buffer; and (iv) dialysis against the modified lysis buffer. Proteins were separated by two-dimensional electrophoresis according to O'Farrell's method. The changes in the silver-stained protein pattern were evaluated by using image processing methods. Electrophoretograms from stressed and nonstressed trees are presented.

Proteins responding to drought and high-temperature stress in Populus × euramericana cv. ‘74/76’

“…Maritime pine (Costa et al 1999;Gion et al 2005), poplar (Renaut et al 2004;Ferreira et al 2006;Plomion et al 2006;Bogeat-Triboulot et al 2007), spruce (Lippert et al 2005;Blödner et al 2007) and oak (Jorge et al 2005(Jorge et al , 2006 are the main forest tree species that have been subjected to extensive proteomic analysis so far. Also, there are some reports on the application of proteomics to the study of responses of forest trees to abiotic stress such as drought (Costa et al 1998;Jorge et al 2006;Plomion et al 2006;Blödner et al 2007;Bogeat-Triboulot et al 2007), ozone (Davidsen 1995) and temperature (Renaut et al 2004;Ferreira et al 2006). The poplar proteomics effort will result in discoveries about the cell's protein machinery that could yield important applications in forestry and lead to an understanding of the molecular basis of poplar responses to abiotic stress.…”

Analysis of bean (Phaseolus vulgaris L.) proteins affected by copper stress

“…Among these inducible plant proteins are pathogenesis related (PR) proteins (Chamnongpol et al, 1996(Chamnongpol et al, , 1998Davidsen, 1995;Kim and Hwang, 2000;Poupard et al, 2001;Przymusiński and Gwóźdź, 1999). Fourteen families of PR-proteins (PR-1 to PR-14) have been previously recognised.…”