The bark is smooth and powdery, pale- or blue-grey to white in colour, with a skirt of rough brownish bark for the bottom 1—4 m of the tree trunk. They are arranged alternately along the branches. The white flowers appear from mid autumn to late winter from April to August. The mountain blue gum E. It is commonly known as the flooded gum and as rose gum in Queensland. Its two closest relatives are the Sydney blue gum Eucalyptus saligna and the mountain blue gum E.

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E-mail: rb. Received Aug 4; Accepted Nov 7. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

This article has been cited by other articles in PMC. Abstract Eucalyptus urograndis is a hybrid eucalyptus of major economic importance to the Brazilian pulp and paper industry. Although widely used in forest nurseries around the country, little is known about the biochemical changes imposed by environmental stress in this species. In this study, we evaluated the changes in the stem proteome after short-term stimulation by exposure to low temperature. Using two-dimensional gel electrophoresis coupled to high-resolution mass spectrometry-based protein identification, 12 proteins were found to be differentially regulated and successfully identified after stringent database searches against a protein database from a closely related species Eucalyptus grandis.

The identification of these proteins indicated that the E. The results of this study represent the first step in understanding the molecular and biochemical responses of E.

Keywords: abiotic stress, mass spectrometry, omics, proteomics Introduction Eucalyptus is one of the most important plant genera used in the pulp and paper industry. Currently, Eucalyptus plants are cultivated worldwide because of their rapid adaptability to different climatic conditions and easy use in plant breeding programs. The most widely used species in forest plantations and breeding programs are Eucalyptus grandis, Eucalyptus globulus, Eucalyptus urophylla and Eucalyptus camaldulensis.

In addition to these species, several intra- and interspecific hybrids that combine important traits from both parental lines have been successfully bred. Eucalyptus urograndis is one of the most important interspecific hybrids because it combines the rapid growth of E. This species is currently the hybrid preferred by the Brazilian pulp industry and is consequently the mostly propagated species in commercial forest nurseries in this country.

Although E. Large-scale gene expression profiling provides a robust, high-throughput means of detecting important genes and of guiding plant breeding programs to improve specific plant traits, such as thermotolerance.

Proteomics, the large-scale study of the proteins present in a particular biological system, is a powerful multi-disciplinary approach that focuses on the characterization of biological molecules that are synthesized as the final product of gene expression. Information regarding the differential regulation of protein s may therefore be useful for monitoring the metabolic responses of plants to environmental disturbances and assisting in the genetic transformation of target plant species.

Despite their importance for the pulp industry, few studies have examined the proteome of Eucalyptusspecies Celedon et al. In this work, we examined the response of young E. Materials and Methods Plant material Young E. Protein extraction After 24 h of cultivation, E. Protein extracts were obtained by mixing 1. After centrifugation, the phenol fraction was collected, transferred to a new tube and five volumes of cold methanol containing mM ammonium acetate was added. Protein quantitation was done with the Bradford assay using bovine serum albumin as the standard.

Vertical electrophoresis was done in a discontinuous buffer system for 2 h at V. Detection of differentially regulated proteins The 2D gels were analyzed using the program Image Master Platinum v. Gel images were acquired in transparent mode using a green color filter.

Spots were detected automatically using the parameters smooth, minimum area and saliency. Spot matching was done by using only one landmark with manual correction if necessary. Supplementary data from the 2D gel image analyses is provided in Table S1. Identification of proteins by tandem mass spectrometry Prior to the mass spectrometric analyses, proteins were digested in gel according to Shevchenko et al.

Mass spectrometric analyses were done in a Q-Exactive instrument Thermo Scientific using the data-dependent acquisition method for the ten most abundant peptide ions. Spectral correlation was done using the Sequest search tool contained in the Proteome Discoverer software Thermo Scientific and run against the E. The search parameters were adjusted for an error tolerance equal to 0. Oxidation of methionines and carbamidomethylation of cysteines were selected as dynamic and static modifications, respectively, during database searches.

Results and Discussion In this study, we used proteomics to obtain information about the E. As expected, there were no major changes in protein accumulation after 24 h of exposure to low temperature.

Using the extraction procedure described here, protein yields of 0. To gain insight into the fine biochemical alterations that could affect growth and wood biosynthesis, the stem proteome was analyzed using 2D gel electrophoresis applied to young E. A subsequent comparative analysis detected 12 protein spots with significant densitometric variation Figure 1 , Table 1.

Tandem mass spectrometric analyses followed by database searches using a closely related Eucalyptusspecies resulted in the successful identification of all proteins from the selected gel spots. The differential regulation of proteins from a wide variety of cellular functions indicates a complex regulatory network in Eucalyptuscells in response to cold stress. Down-regulation of the proteins fructose bisphosphate aldolase Eucgr.


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