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.

Recebido para publicação em 03/09/2013 Aceito para publicação em 28/05/2014 RESUMO. O fruto de noni (Morinda citrifolia L.) é consumido há milênios na medicina popular polinésia devido aos benefícios nutricionais e terapêuticos. O consumo de noni em outros países, incluindo o Brasil, cresceu vertiginosamente nos últimos anos em decorrência das atividades biológicas atribuídas a ingestão do suco da fruta, principalmente pela propriedade anticâncer. Contudo, a composição química da planta, que está relacionada com suas propriedades biológicas, é determinada pelo seu local de origem, e por influência do clima e do solo onde é cultivada. Neste sentido, este trabalho teve como objetivo analisar a polpa extraída de frutos maduros de noni cultivados em Maringá-PR. A análise da polpa in natura apresentou 89,16% ABSTRACT: Proximate composition, bioactive compounds and antioxidant activity of fruits of Morinda citrifolia L. (noni) cultivated in Paraná, Brazil. The noni fruit (Morinda citrifolia L.) has been consumed for millennia in the Polynesian folk medicine because of its nutritional and therapeutic benefits. The consumption of the noni fruit in other countries, including Brazil, has increased in recent years because of the biological activities attributed to the ingestion of noni juice, especially the anticancer effect. However, the chemical composition of the plant, which is related to its biological properties, is determined by its geographical origin and is influenced by climate and soil. Therefore, this study aimed to analyze the pulp extracted from noni fruit grown in Maringá, state of Paraná, Brazil. The fresh pulp analysis showed 89.16% of moisture, 0.75% of ash, 2.10% of protein, 2.19% of fat and 5.81% of carbohydrates. The bioactive compounds analyzed were anthocyanins (1.39 mg.100 g -1 pulp), yellow flavonoids (13.01 mg.100 g -1 pulp), carotenoids (0.45 mg.100 g -1 pulp) and vitamin C (12.16 mg.100 g -1 pulp). For the analysis of total phenols and antioxidant activity, different extracts were prepared. The highest total phenolic contents were found in the aqueous extract (1143.56 mg EAG.100 g -1 ), followed by the ethanol extract (966.96 mg EAG.100 g -1 ), methanol/acetone extract (820.88 mg EAG.100 g -1 ) and methanol extract (306.33 mg EAG.100 g -1 ). The results for the antioxidant capacity were determined by the EC 50 , which is concentration of extract required to reduce 50% of the DPPH radical. The best results for the antioxidant capacity were found in the methanol/ acetone extract (EC 50 of 25.18 mg.mL -1 ) and in the methanol extract (EC 50 of 25.96 mg.mL -1 ). The antioxidant activity of the fruit may be related to its vitamin C content, since the extracts with lower total phenolic content were those that had lower EC 50 values.

PCR analysis of tRNA intergenic spacer (tDNA-PCR) and of the 16S-23S internal transcribed spacer (ITS-PCR) and random amplified polymorphic DNA (RAPD) analysis were evaluated for their usefulness in characterization of Enterobacter cloacae strains isolated from both clinical origins and vaccine microbial contamination. tDNA-PCR presented specific and reproducible patterns for Enterobacter sakazakii ATCC 29004, Enterobacter aerogenes ATCC 13048, and Enterobacter cloacae ATCC 13047 and 23355 that presented the same profile for all 16 E. cloacae isolates, offering an alternative tool for species-level identification. ITS-PCR and RAPD analysis yielded completely different banding patterns for the 20 strains studied, except for E. cloacae strains isolated from different batches of vaccine that exhibited a unique pattern, suggesting contamination by the same strain. The combined use of tDNA-PCR and ITS-PCR in a one-step protocol allows accurate identification and typing of E. cloacae strains a few hours after the colony has been isolated.

Tabernaemontana catharinensis A. DC (Apocynaceae) is used as a medicinal plant by the population. In order to contribute to the safe use of the plant as herbal medicine, this study aimed to morphoanatomically characterize the aereal vegetative organs of T. catharinensis and to evaluate the leaves' mutagenic and antimutagenic activities. Histological blades of leaves and stem of T. catharinensis were performed; the methionine system (methG1) and Aspergillusnidulans conidia germination analysis were employed for mutagenic and antimutagenic evaluation. The morphoanatomic analysis did not show trichomes in the stem, petiole and leaf. Besides, it was observed both the presence of bi-collateral bundles -except in the foliar apex where the bundles were from the collateral type -as well as anamphistomatic leaf with paracyte stomata and sub-epidermal layer in the region of the leaf edges. The mutagenicity/ antimutagenicity trial indicated a significant decrease of mutation frequency in comparison with the control group and showed that theT. catharinensis had antimutagenic activity within the type, time and form of treatment. Since the germination test showed that the conidia germination was accelerated from the bud phase, activities at the cell cycle level and polarized growth proved to be possible. The morphoanatomic analysis of the leaf and stem associated with the mutagenic and antimutagenic analyses contributes to the safe use of the plant by humans and also for the quality control of a possible phytotherapeutic drug.Keywords: Aspergillus nidulans, genotoxicity, medicinal herbs, methionine system, morphoanatomy.RESUMO: Caracterização morfoanatômica do caule e da folha de Tabernaemontana catharinensis A.DC (Apocynaceae) e atividade antimutagênica de suas folhas Tabernaemontana catharinensis A. DC (Apocynaceae) é utilizada como planta medicinal pela população. A fim de contribuir para o uso seguro da planta como medicinal, este trabalho teve como objetivo caracterizar morfoanatomicamente os órgãos vegetativos aéreos de T. catharinensis e avaliar a atividade mutagênica e antimutagênica de suas folhas. Foram realizados cortes histológicos da folha e do caule de T. catharinensis e, para a avaliação mutagênica e antimutagênica, foi utilizado o sistema metionina (methG1) e análise da germinação de conídios em Aspergillus nidulans. A análise morfoanatômica evidenciou a ausência de tricomas no caule, pecíolo e folha; presença de feixes bicolaterais, com exceção no ápice foliar cujos feixes são do tipo colateral; folha anfiestomática com estômatos paracíticos e camada subepidérmica na região do bordo foliar. O ensaio de mutagenicidade/antimutagenicidade mostrou uma diminuição significativa da frequência de mutação em relação ao controle, indicando que nesse tipo, tempo e forma de tratamento, T. catharinensis apresentou atividade antimutagênica. O ensaio de germinação evidenciou que houve aceleração da germinação dos conídios, a partir da fase de botão, indicando uma possível atuação em nível de ativação de ciclo celu...

A tropical endophytic isolate of the fungus Fusarium verticillioides (synonym Fusarium moniliforme) obtained from Zea mays was co-transformed with plasmid pNH24 containing the Fusarium oxysporum nitrate reductase nia gene and plasmid pNOM 102 carrying the Escherichia coli β-glucuronidase gusA gene. Transformation frequency for the nia marker was 75 transformants µg -1 vector DNA and introduction of the gusA gene by co-transformation was 57.2% as indicated by the presence of the GUS + phenotype on plates. Southern analyses confirmed the integration of both plasmids into the genome of ten GUS + transformants. All co-transformants showed mitotic stability in respect of the GUS + phenotype. To assess the potential of transformed endophytic fungi as vectors for introducing desirable characteristics into host tropical plants of biotechnological and agricultural importance we successfully infected maize roots and detected GUS + phenotype".

ABSTRACT. Apoptosis and necrosis are among several types of cell death. We stained the nuclei of Aspergillus nidulans grown in microcolonies with ethidium bromide and acridine orange to detect in situ apoptosis. Suspensions of conidia from 5-day-old colonies of the A. nidulans strains biA1methG1, G422, CLC100, and CLB3 were each put into two tubes. The suspension of one tube was irradiated with ultraviolet light for 20 s, whereas the other tube was not exposed to irradiation. The two suspensions were inoculated in complete liquid medium and 50-µL samples were placed on sterilized cover slips, spread on the surface of solid culture media on Petri dishes. After the micro-colonies were formed, the material on the cover slips was stained with ethidium bromide and acridine orange, placed on the lamina and observed under a fluorescence microscope. This staining method was efficient in discriminating normal nuclei from those going apoptosis and necrosis. Results have shown that irradiation provokes apoptosis but does not induce necrosis. There were no differences between the three strains and all data were considered to be statistically significant.

ABSTRACT. Grape juice, in addition to being an energetic food, due to its high sugar content, has several compounds that can prevent or treat various types of diseases. Resveratrol is a compound present in grapes that has attracted a lot of interest; in addition to preventing cardiovascular disease linked to lipid metabolism, it has chemopreventive and chemotherapeutic activities. We evaluated the antimutagenic activity and determined the trans-resveratrol content in grape juice from the varieties Vênus, BRS Violeta and Isabel. The grape juices from the three cultivars and the resveratrol solution were tested in the methG1 system in Aspergillus nidulans. The conidia from the biA1methG1 strain were treated for 4 h in 10% grape juice (v/v). After washing, the conidia were placed in selective media to analyze survival and mutations. The standard resveratrol solution and the grape juice of the cultivar Isabel, both with a trans-resveratrol content of 1 mg/ mL, presented antimutagenic potential in this test system because the frequency of mutation of the treatments was significantly lower than the frequency of spontaneous mutation. However, grape juice from Evaluation of antimutagenicity of grape juice the varieties Vênus and BRS Violeta, both with a lower quantity of trans-resveratrol, gave weak antimutagenic activity in this test system because the frequency of mutation of the treatments was significantly higher than the frequency of spontaneous mutation.

ABSTRACT. Medicinal plants such as Aloe arborescens Miller andAloe barbadensis Miller are used by the general population to treat various diseases. Therefore, the aim of this study was to evaluate the antimutagenicity of these two species using a methG1 system in Aspergillus nidulans and the comet assay in rats. The animals were treated with the plants at concentrations of 360 and 720 mg/kg body weight (1 and 2, respectively) by gavage for 14 days, followed by the administration of etoposide on treatment day 8. Blood samples were prepared for analysis of DNA damage. For the test in A. nidulans, the biA1methG1 lineage conidia were treated for 4 h with both plant species at concentrations of 4 and 8% (w/v). Then, they were washed and plated on a selective medium for frequency analysis of survival and mutation. The results of the comet assay showed that both plants were antigenotoxic compared to etoposide, which was not a typical response of methG1 systems, where only the highest concentration of plant extracts usually exhibit beneficial effects. This study demonstrates the potential antigenotoxicity and antimutagenicity of the Aloe plants tested and, therefore, supports their use as a form of preventive therapy and for health maintenance by the population.