Recent advances in genomics and proteomics have provided an excellent opportunity to understand complex biological processes such as wood formation at the gene and protein levels. The aim of this work was to describe the proteins participating in the processes involved in juvenile wood formation by isolating proteins from the cambial region of Eucalyptus grandis, at three ages of growth (6-month-old seedlings, 3- and 6-year-old trees), and also to identify proteins differentially expressed. Using a 2-D-LC-MS/MS strategy we identified a total of 240 proteins, with 54 corresponding spots being present in at least two ages. Overall, nine proteins classified into the functional categories of metabolism, cellular processes, and macromolecular metabolism showed significant changes in expression. Proteins were classified into seven main functional categories, with metabolism representing 35.2% of the total proteins identified. The comparison of the reference maps showed not only differences in the expression pattern of individual proteins at each age, but also among isoforms. The results described in this paper provide a dynamic view of the proteins involved in the formation of juvenile wood in E. grandis.
Despite the importance of Eucalyptus spp. in the pulp and paper industry, functional genomic approaches have only recently been applied to understand wood formation in this genus. We attempted to establish a global view of gene expression in the juvenile cambial region of Eucalyptus grandis Hill ex Maiden. The expression profile was obtained from serial analysis of gene expression (SAGE) library data produced from 3- and 6-year-old trees. Fourteen-base expressed sequence tags (ESTs) were searched against public Eucalyptus ESTs and annotated with GenBank. Altogether 43,304 tags were generated producing 3066 unigenes with three or more copies each, 445 with a putative identity, 215 with unknown function and 2406 without an EST match. The expression profile of the juvenile cambial region revealed the presence of highly frequent transcripts related to general metabolism and energy metabolism, cellular processes, transport, structural components and information pathways. We made a quantitative analysis of a large number of genes involved in the biosynthesis of cellulose, pectin, hemicellulose and lignin. Our findings provide insight into the expression of functionally related genes involved in juvenile wood formation in young fast-growing E. grandis trees.
Neuroendocrine tumours are a heterogeneous group of diseases with a significant variety of diagnostic tests and treatment modalities. Guidelines were developed by North American and European groups to recommend their best management. However, local particularities and relativisms found worldwide led us to create Brazilian guidelines. Our consensus considered the best feasible strategies in an environment involving more limited resources. We believe that our recommendations may be extended to other countries with similar economic standards.
A method for genetic transformation of germinating seeds and seedlings of
Eucalyptus grandis ×
E. urophyllais described using the sonication-assisted
Agrobacterium-mediated transformation (SAAT) system.
Seeds germinated for 2 d, and 15-d-old seedlings, sonicated for 30 s, had the
highest percentage of β-glucuronidase (GUS) transient expression (21.7 and
37.4%, respectively). Pre-sonication greatly enhanced the efficiency of
transformation. The differential transformation of tissues was also
investigated, with seeds imbibed for 2 d having over 90% of the blue
sectors localised in cotyledons and in the intersection of the hypocotyls and
roots, whereas in 5-d-old seedlings, 70% of GUS activity was detected
in cotyledons. However, 15–17-d-old seedlings had around 60% of
transformed sectors localised in the first pair of leaves. The efficiency of
the method was also assessed using a chimeric construct containing the
Lhcb1*2 gene of the 28 kDa chlorophyll
a/b binding pea protein from the LHCII antenna. Four
stable transformants were confirmed by genomic blotting.
Our results characterized the changes in the proteome of P. gingivalis following exposure to nicotine and cotinine, suggesting that these substances may modulate, with minor changes, protein expression. The present study is, in part, a step toward understanding the potential smoke-pathogen interaction that may occur in smokers with periodontitis.
Volatile organic compounds (VOCs) released by Saccharomyces cerevisiae inhibit plant pathogens, including the filamentous fungus Phyllosticta citricarpa, causal agent of citrus black spot. VOCs mediate relevant interactions between organisms in nature, and antimicrobial VOCs are promising, environmentally safer fumigants to control phytopathogens. As the mechanisms by which VOCs inhibit microorganisms are not well characterized, we evaluated the proteomic response in P. citricarpa after exposure for 12h to a reconstituted mixture of VOCs (alcohols and esters) originally identified in S. cerevisiae. Total protein was extracted and separated by 2D-PAGE, and differentially expressed proteins were identified by LC-MS/MS. About 600 proteins were detected, of which 29 were downregulated and 11 were upregulated. These proteins are involved in metabolism, genetic information processing, cellular processes, and transport. Enzymes related to energy-generating pathways, particularly glycolysis and the tricarboxylic acid cycle, were the most strongly affected. Thus, the data indicate that antimicrobial VOCs interfere with essential metabolic pathways in P. citricarpa to prevent fungal growth.
The aryloxyphenoxypropionate herbicides (APPs) are graminicides with excellent control of many grass weeds species, including weedy rice (Oryza sativa L.). These herbicides block the fatty acid biosynthesis by inhibiting the enzyme acetyl-CoA carboxylase (ACCase), resulting in the death of susceptible plants. Inducing mutation with gamma rays to rice seeds, two lines resistant to APPs herbicides were developed. Plant dose-response assays confirmed the resistance to the APPS herbicides quizalofop-p-ethyl and haloxyfop-p-methyl. The carboxyltransferase domain fragments of ACCase from the resistant biotype and susceptible control were sequenced and compared. A point mutation was detected in the amino acid position 2,027 (Rice Genome Annotation Project: Os05g22940.1). Results indicated that resistance to APPs is a consequence of an altered ACCase enzyme that confers resistance. The use of APPs herbicide-resistant rice lines represents an innovative and promising alternative for weedy rice control in paddy rice systems.
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