The molecular mechanisms of plant recognition, colonization, and nutrient exchange between diazotrophic endophytes and plants are scarcely known. Herbaspirillum seropedicae is an endophytic bacterium capable of colonizing intercellular spaces of grasses such as rice and sugar cane. The genome of H. seropedicae strain SmR1 was sequenced and annotated by The Paraná State Genome Programme—GENOPAR. The genome is composed of a circular chromosome of 5,513,887 bp and contains a total of 4,804 genes. The genome sequence revealed that H. seropedicae is a highly versatile microorganism with capacity to metabolize a wide range of carbon and nitrogen sources and with possession of four distinct terminal oxidases. The genome contains a multitude of protein secretion systems, including type I, type II, type III, type V, and type VI secretion systems, and type IV pili, suggesting a high potential to interact with host plants. H. seropedicae is able to synthesize indole acetic acid as reflected by the four IAA biosynthetic pathways present. A gene coding for ACC deaminase, which may be involved in modulating the associated plant ethylene-signaling pathway, is also present. Genes for hemagglutinins/hemolysins/adhesins were found and may play a role in plant cell surface adhesion. These features may endow H. seropedicae with the ability to establish an endophytic life-style in a large number of plant species.

This study was funded by IVF-Spain Foundation. The authors have no conflicts of interest to declare.

The aim of this study was to characterise Discrete Typing Units (DTUs) of 28 isolates of Trypanosoma cruzi from humans (15), triatomines (9), and opossums (4) in the state of Paraná, southern Brazil. For this purpose, we analysed the size polymorphism at the 3' end of the 24Sα ribosomal RNA gene (rRNA) and the restriction fragment length polymorphism (RFLP) of the partial 5' sequence of the mitochondrial Cytochrome Oxidase subunit II gene (COII). Band patterns of the isolates were compared with reference samples of T. cruzi I (Silvio X10 and Col 17G2), T. cruzi II (Esmeraldo and JG), T. cruzi III (222 and 231), T. cruzi IV (CAN III), T. cruzi V (SO3 cl5), and T. cruzi VI (CL Brener). Our results confirmed that rRNA analysis is of limited use for assessing T. cruzi DTUs. COII RFLP analysis was suitable for screening, but for one isolate it was necessary to determine the COII partial sequence to identify the DTU. Only one of the isolates from humans belonged to T. cruzi I; 13 isolates belonged to T. cruzi II and one to T. cruzi III. The four isolates from opossums and five isolates from triatomines were identified as T. cruzi I. Four isolates from triatomines showed patterns of both T. cruzi I and II, indicating mixed infections. This study contributes to the characterisation of the dynamics of T. cruzi populations in southern Brazil.

We have identified bent DNA sites in the distal and proximal DNA puff BhC4-1 amplified gene promoter region of Bradysia hygida. The 2D modeling of the 3D DNA path and the ENDS ratio values calculated in this promoter region resulted in the identification of ten pronounced bent sites named BhC4B - 9 to + 1. The 1847 bp fragment (- 3697 to - 1850) in relation to the transcription start site shows multiple bending sites, BhC4B - 9 to BhC4B - 4, with periodicity approximately 300 bp. The analysis of the other identified bent region, starting at position - 957, reveals that the BhC4B + 1 bent site colocalizes with the putative BhC4-1 minimal promoter. The sequence analysis of bent site BhC4B - 4 shows a distribution of dA*dT at approximately 10 bp intervals between the middle of each tract, but intervals with more than one turn, approximately 20 bp, two helix turns, were detected in the other bent sites described here. The bent sites BhC4B - 6 and BhC4B - 4, contain two consensus sequences, with 60 bp each. The apparent molecular weight of fragments in the BhC4-1 promoter region were estimated in agarose gels and compared with the data obtained in polyacrylamide gels without and with ethidium bromide. The mobility reduction ratios (R-values) were determined, and a high R-value, 1.80, for a 1215 bp fragment in the distal promoter region and a 1.23 significant R-value for a 662 bp fragment in the proximal segment were found. To further analyze the predicted bent DNA sites in these fragments, the 2D trajectories of the 3D DNA path and other parameters, AT percentage, roll angle, ENDS ratio and DeltaG, were determined. The role of these bent sites in the BhC4-1 transcription regulation is discussed.

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