BackgroundGluconacetobacter diazotrophicus Pal5 is an endophytic diazotrophic bacterium that lives in association with sugarcane plants. It has important biotechnological features such as nitrogen fixation, plant growth promotion, sugar metabolism pathways, secretion of organic acids, synthesis of auxin and the occurrence of bacteriocins.ResultsGluconacetobacter diazotrophicus Pal5 is the third diazotrophic endophytic bacterium to be completely sequenced. Its genome is composed of a 3.9 Mb chromosome and 2 plasmids of 16.6 and 38.8 kb, respectively. We annotated 3,938 coding sequences which reveal several characteristics related to the endophytic lifestyle such as nitrogen fixation, plant growth promotion, sugar metabolism, transport systems, synthesis of auxin and the occurrence of bacteriocins. Genomic analysis identified a core component of 894 genes shared with phylogenetically related bacteria. Gene clusters for gum-like polysaccharide biosynthesis, tad pilus, quorum sensing, for modulation of plant growth by indole acetic acid and mechanisms involved in tolerance to acidic conditions were identified and may be related to the sugarcane endophytic and plant-growth promoting traits of G. diazotrophicus. An accessory component of at least 851 genes distributed in genome islands was identified, and was most likely acquired by horizontal gene transfer. This portion of the genome has likely contributed to adaptation to the plant habitat.ConclusionThe genome data offer an important resource of information that can be used to manipulate plant/bacterium interactions with the aim of improving sugarcane crop production and other biotechnological applications.

Metallo-beta-lactamase production is emerging worldwide as an important mechanism of carbapenem resistance among nonfermentative Gram-negative isolates, and this mechanism is becoming frequently observed in Brazil. This study documents the occurrence and characteristics of an epidemic SPM-1-producing Pseudomonas aeruginosa strain in a teaching hospital located in Rio de Janeiro City, Brazil. The bla (SPM-1) gene and a class 1 integron were detected in 13 isolates, representing 20% of the 65 imipenem-resistant P. aeruginosa isolates obtained from January, 2000, to August, 2001. DNA sequencing revealed that this integron carries three gene cassettes that confer resistance to antimicrobials, aacA4, bla (OXA-56), and aadA7, and an orf1 encoding a putative transposase. All 13 SPM-producing P. aeruginosa isolates had closely related pulsed-field gel electrophoresis (PFGE) profiles, designated as clonal group A, suggesting nosocomial spread of the strain. This clonal group was the same as that observed in other SPM-1-producing P. aeruginosa isolates from distinct Brazilian states. The dissemination of this clone throughout Brazil could not be explained by transfer of infected patients and/or sharing of common health-care staff. It is likely that the spread of these strains occurred indirectly via the community.

The regeneration patterns of shoot apices derived from in vitro plants of four varieties of sugar cane in response to different growth regulators and light were evaluated. The cellular origin of the regeneration processes was also investigated. Explants cultivated on medium supplemented with NAA and incubated under light showed direct bud regeneration from cells of external layers of the ground parenchyma of the stem. Explants cultivated in the dark on medium supplemented with low concentrations of picloram (PIC) or 2,4D (4.0 and 4.5 mM, respectively) showed callus formation derived from the ground parenchyma of stem and development of preembryogenic masses derived from bundle sheath cells facing the phloem tissue of immature leaves. Somatic embryos at further developmental stages were visible following transfer to medium devoid of growth regulators and incubation under light. When incubated under light since the begining of the experiment, explants cultivated in the presence of higher PIC or 2,4D concentrations (40 and 22.6 mM, respectively) first displayed direct organogenesis from external layers of the ground parenchyma of the stem, followed by the development of organogenic calluses. Preembryogenic masses were also observed from bundle sheath cells of immature leaves. However, in contrast to the cultures pre-incubated in darkness for 30 days, the subsequent stages of embryo development were not detected. The regeneration efficiency of calluses induced by 2,4D and PIC was generally increased following desiccation in laminar flow or incubation on medium solidified with phytagel.

Resumo -O objetivo deste trabalho foi estabelecer sistemas de multiplicação de plantas de cana-de-açúcar in vitro e avaliar sua utilização, como material inicial, para a indução de regeneração a partir de ápices caulinares. Três métodos de cultivo foram avaliados: cultura em meio semi-sólido, cultura líquida estacionária e cultura líquida sob agitação. A taxa de multiplicação mais elevada foi alcançada por meio da cultura líquida sob agitação. Ápices caulinares, excisados dessas plantas, apresentaram taxas de regeneração in vitro compatíveis com sua utilização em protocolos de transformação. Calos resistentes a PPT e GUS-positivos foram obtidos de explantes da variedade Chunnee com inoculação de Agrobacterium tumefaciens C58C1 (pMP90) (pDUBarA9). O protocolo estabelecido a partir de cultivo in vitro pode ser utilizado para a produção de plantas transgênicas de cana-deaçúcar, visando à realização de estudos de regulação da expressão gênica, assim como à introdução de características de interesse agronômico.Termos para indexação: Saccharum, micropropagação, transformação genética, Agrobacterium tumefaciens. In vitro morphogenesis of Brazilian sugarcane varietiesAbstract -The objective of this work was to establish in vitro systems for sugarcane plant multiplication and for regeneration from shoot apices excised from these plants. Three methods were analyzed: culture on semi-solid medium and liquid culture with or without agitation. The highest regeneration rates were obtained from cultures in liquid-medium with agitation. Shoot tips derived from these plants presented regeneration rates suitable for utilization in transformation protocols. PPT-resistant and GUS positive calluses were obtained from explants of in vitro plants of Chunnee variety inoculated with Agrobacterium tumefaciens C58C1 (pMP90) (pDUBarA9). The established in vitro culture system can be applied for transgenic sugarcane production, aiming at gene regulation studies and introduction of agronomical traits.

Oesophageal squamous cell carcinoma (ESCC) is one of the most common malignancies and is the sixth cause of cancer-related death in the world. Inactivation of cell-cycle regulating genes, such as p14ARF and p16INK4a, and cell adhesion genes, such as E-cadherin, is common in cancer, and results from genetic and/or epigenetic alterations. Therefore, we have analysed the mRNA expression of p14ARF, p16INK4a and E-cadherin in 17 matched ESCC and normal mucosal samples obtained from Brazilian patients by semi-quantitative RT-PCR. The expression of p14ARF and p16INK4a was absent or reduced in several ESCC samples. Hypermethylation of CpG islands, caused by the action of DNA methyltransferases (DNMTs), is a major form of epigenetic inactivation of the p14ARF and p16INK4a genes in tumours. Hence, we also investigated the mRNA expression of the human DNA methyltransferases in normal oesophageal mucosa and in the tumour matched samples. All DNMTs were constitutively expressed in the normal oesophageal mucosa but a significantly higher expression of DNMT3B was observed in the tumours. Data analysis by the Spearman rank test showed that the expression of DNMT3B was inversely correlated with that of p14ARF and p16INK4a. Our results suggest that DNMT3B over-expression may be involved in the suppression or lower expression of p14ARF and p16INK4a observed in ESCC.