Lippia origanoides essential oil: an efficient and safe alternative to preserve food, cosmetic and pharmaceutical products.

AIMS: The aim of this work was to evaluate the efficacy and safety of Lippia origanoides essential oil as a preservative in industrial products. METHODS AND RESULTS: The composition, antimicrobial activity, mutagenic and toxic potential of L. origanoides were determined. Then, the effect of essential oil as a preservative in food, cosmetics and pharmaceutical products was evaluated. The essential oil of L. origanoides consisted mainly of oxygenated monoterpenes (38·13%); 26·28% corresponded to the compound carvacrol. At concentrations ranging from 0·312 to 1·25 µl ml-1 and in association with polysorbate 80, the essential oil of L. origanoides inhibited the growth of all the tested micro-organisms. The medium lethal dose in mice was 3·5 g kg-1 , which categorizes it as nontoxic according to the European Union criteria, and negative results in the Ames test indicated that this oil was not mutagenic. In combination with polysorbate 80, the essential oil exerted preservative action on orange juice, cosmetic and pharmaceutical compositions, especially in the case of aqueous-based products. CONCLUSIONS: Lippia origanoides essential oil is an effective and safe preservative for orange juice, pharmaceutical and cosmetic products. SIGNIFICANCE AND IMPACT OF THE STUDY: This study allowed for the complete understanding of the antimicrobial action and toxicological potential of L. origanoides essential oil. These results facilitate the development of a preservative system based on L. origanoides essential oil.

Genetic diversity analysis of Croton antisyphiliticus Mart. using AFLP molecular markers.

Croton antisyphiliticus Mart. is a medicinal plant native to Cerrado vegetation in Brazil, and it is popularly used to treat urogenital tract infections. The objective of the present study was to assess the genetic variability of natural C. antisyphiliticus populations using AFLP molecular markers. Accessions were collected in the states of Minas Gerais, São Paulo, and Goiás. The genotyping of individuals was performed using a LI-COR® DNA Analyzer 4300. The variability within populations was found to be greater than the variability between them. The F(ST) value was 0.3830, which indicated that the populations were highly structured. A higher percentage of polymorphic loci (92.16%) and greater genetic diversity were found in the population accessions from Pratinha-MG. Gene flow was considered restricted (N(m) = 1.18), and there was no correlation between genetic and geographic distances. The populations of C. antisyphiliticus exhibited an island-model structure, which demonstrates the vulnerability of the species.

Improving plant transformation using Agrobacterium tumefaciens.

Here, we report a quick and low-cost method to improve plant transformation using Agrobacterium tumefaciens. This method involves the use of physical wounding, ultrasound, and an increase in exposure time to the bacteria. We show how the transformation rate increased from 0 to 14% when an ultrasound pulse of 10 s was used in conjunction with 96 h of bacterial exposure in Eclipta alba explants.

Enraizamento in vitro de catuaba (Anemopaegma arvense (Vell. ) Stell. ex de Souza), uma planta medicinal do Cerrado / Study on the in vitro rooting of Anemopaegma arvense (Vell. ) Stell. ex de Souza, a medicinal plant of the Brazilian Cerrado

Este trabalho foi realizado com o objetivo de otimizar o protocolo para enraizamento in vitro de Anemopaegma arvense, planta medicinal do Cerrado em risco de extinção e conhecida popularmente como catuaba, a qual é amplamente utilizada na medicina popular. Brotações cultivadas in vitro foram inoculadas em meio de cultura MS/2 liquido e MS sólido suplementado com diferentes concentrações de auxinas, poliaminas ou dithiothreitol (DTT). As avaliações foram realizadas quanto à porcentagem de enraizamento, número e comprimento das raízes. A presença de NAA (Ácido naftaleno acético) no meio de cultura foi essencial para promover a indução de raízes adventícias nas brotações. A maior porcentagem de enraizamento, 50%, foi obtida no tratamento 2 mg L-1 de NAA com tempo de permanência de 15 dias nesta auxina. No experimento com poliaminas o melhor tratamento foi MS/2 + 5 mg L-1 de putrescina, com 27% de brotações enraizadas. Na presença de DTT (Dithiothreitol), 23% das brotações enraizaram em 0,10 mg L-1 de DTT. A presença da auxina NAA e a alternância no tempo de permanência foi a melhor condição para promover o enraizamento in vitro da de A. arvense. .

This work was carried out in order to optimize an efficient protocol for the in vitro rooting of Anemopaegma arvense, a medicinal plant of the Brazilian Cerrado in danger of extinction, popularly known as Catuaba in Portuguese and widely used in folk medicine. Shoots cultivated in vitro were inoculated in liquid MS/2 and solid MS culture medium supplemented with different concentrations of auxins, polyamines or dithiothreitol (DTT). Evaluations were performed for the rooting percentage and for the number and length of roots. The presence of NAA (naphthaleneacetic acid) in the culture medium was essential to promote the induction of adventitious roots. Higher rooting percentage (50%) was obtained in the treatment with 2 mg L-1 NAA and duration of stay of 15 days in this auxin. In the experiment with polyamines, the best treatment was MS/2 + 5 mg L-1 putrescine with 27% of shoots rooted. In the presence of DTT (dithiothreitol), 23% of shoots rooted at 0.10 mg L-1 DTT. The presence of the auxin NAA and the alternation in length of stay was the best condition to promote in vitro rooting of A. arvense. .

Search for methylation-sensitive amplification polymorphisms in mutant figs.

Fig (Ficus carica) breeding programs that use conventional approaches to develop new cultivars are rare, owing to limited genetic variability and the difficulty in obtaining plants via gamete fusion. Cytosine methylation in plants leads to gene repression, thereby affecting transcription without changing the DNA sequence. Previous studies using random amplification of polymorphic DNA and amplified fragment length polymorphism markers revealed no polymorphisms among select fig mutants that originated from gamma-irradiated buds. Therefore, we conducted methylation-sensitive amplified polymorphism analysis to verify the existence of variability due to epigenetic DNA methylation among these mutant selections compared to the main cultivar 'Roxo-de-Valinhos'. Samples of genomic DNA were double-digested with either HpaII (methylation sensitive) or MspI (methylation insensitive) and with EcoRI. Fourteen primer combinations were tested, and on an average, non-methylated CCGG, symmetrically methylated CmCGG, and hemimethylated hmCCGG sites accounted for 87.9, 10.1, and 2.0%, respectively. MSAP analysis was effective in detecting differentially methylated sites in the genomic DNA of fig mutants, and methylation may be responsible for the phenotypic variation between treatments. Further analyses such as polymorphic DNA sequencing are necessary to validate these differences, standardize the regions of methylation, and analyze reads using bioinformatic tools.

Ocorrência de fungos micorrízicos em catuaba (Anemopaegma arvense (Vell. ) Stell. ex de Souza - Bignoniaceae), uma planta medicinal do Cerrado em risco de extinção / Michorrizical funghi studies in catuaba (Anemopaegma arvense (Vell. ) Stell. ex de Souza), a medicinal plant from the Cerrado region at risk of extinction

Este trabalho foi realizado com o objetivo de verificar a ocorrência de fungos micorrízicos, identificar suas espécies, e avaliar a colonização em raízes de plantas de diferentes populações e variedades de Anemopaegma arvense, uma planta medicinal do Cerrado em risco de extinção. As avaliações da colonização micorrízica e identificação de espécies de fungos micorrízicos arbusculares (FMAs) foram realizadas no Laboratório de Microbiologia do Solo da Universidade Federal de Lavras (UFLA). Foram utilizadas raízes de 10 plantas coletadas em diferentes locais juntamente com as amostras de solo próximo ao sistema radicular de populações naturais das variedades: glabra, puberula e petiolata. Os esporos foram extraídos do solo por meio da metodologia de peneiramento úmido e, para a avaliação e observação da colonização radicular, as raízes coradas foram observadas em microscópio estereoscópico (ampliação 10 a 40x). Em todas as amostras analisadas foi possível identificar espécies de FMAs, exceto para a variedade puberula, coletada no município de Mogi Guaçu-SP. O número de esporos variou de maneira significativa entre as amostras e os locais amostrados, não havendo distribuição regular. Acaulospora scrubiculata, A. spinosa, A. longula, Escustelospora heterogama, Paraglomus occultum, Gigaspora margarita, Gigaspora sp., dentre outros, encontram-se entre as espécies identificadas. Quanto à avaliação da colonização micorrízica nas raízes, observou-se que todas as variedades foram colonizadas no sitema radicular, verificada por meio da presença de hifas. No entanto, a colonização não foi constatada em todas as amostras avaliadas e também não foi observada a formação de arbúsculos e/ou vesículas.

This work was carried out to verify the occurrence, identify species of mycorrhizal fungi and to evaluate the colonization in roots of plants of different varieties and populations of Anemopaegma arvense, a medicinal plant from the Savannah, in danger of extinction. Assessments of mycorrhizal infection and identification of species of mycorrhizal fungi (AMF) were conducted at the Laboratory of Soil Microbiology, Federal University of Lavras (UFLA). We used roots from 10 plants collected at different sites, along with samples of soil near the root system of natural populations of the varieties glabra, puberula and petiolata. Spores were extracted from soil by the wet sifting methodology and, for the assessment and observation of the root colonization, the stained roots were observed under a stereomicroscope (magnification 10 to 40x), respectively. In all samples it was possible to identify AMF species, except for the variety puberula, collected in the municipality of Mogi Guaçu - SP. The number of spores varied significantly between samples and sampling sites, with no regular distribution. Acaulospora scrubiculata, A. spinosa, A. longula, Escustelospora heterogama, Paraglomus occultum, Gigaspora margarita, Gigaspora sp. among others, are some of the species identified. Regarding the assessment of mycorrhizal infection in the roots, we observed that all the varieties analyzed showed colonization by hyphae. However, not all samples assessed presented colonization and the formation of arbuscules and/or vesicles was not observed.

Genetic characterization of fig tree mutants with molecular markers.

The fig (Ficus carica L.) is a fruit tree of great world importance and, therefore, the genetic improvement becomes an important field of research for better crops, being necessary to gather information on this species, mainly regarding its genetic variability so that appropriate propagation projects and management are made. The improvement programs of fig trees using conventional procedures in order to obtain new cultivars are rare in many countries, such as Brazil, especially due to the little genetic variability and to the difficulties in obtaining plants from gamete fusion once the wasp Blastophaga psenes, responsible for the natural pollinating, is not found in Brazil. In this way, the mutagenic genetic improvement becomes a solution of it. For this reason, in an experiment conducted earlier, fig plants formed by cuttings treated with gamma ray were selected based on their agronomic characteristics of interest. We determined the genetic variability in these fig tree selections, using RAPD and AFLP molecular markers, comparing them to each other and to the Roxo-de-Valinhos, used as the standard. For the reactions of DNA amplification, 140 RAPD primers and 12 primer combinations for AFLP analysis were used. The selections did not differ genetically between themselves and between them and the Roxo-de-Valinhos cultivar. Techniques that can detect polymorphism between treatments, such as DNA sequencing, must be tested. The phenotypic variation of plants may be due to epigenetic variation, necessitating the use of techniques with methylation-sensitive restriction enzymes.

Conservação e enraizamento in vitro de infalível (Mandevilla velutina K. Schum.), uma planta medicinal do Cerrado / In vitro conservation and rooting of "infalível" (Mandevilla velutina K. Schum.), a medicinal plant of Cerrado

Mandevilla velutina (Apocynaceae) é uma planta medicinal endêmica do Cerrado brasileiro conhecida popularmente como infalível, utilizada pela população em tratamentos de processos inflamatórios e acidentes com serpentes. Atualmente, esta espécie encontra-se em risco de extinção, devido à coleta extrativista. Os objetivos deste trabalho foram otimizar o protocolo para o enraizamento in vitro de M. velutina e introduzir diferentes genótipos em banco de germoplasma in vitro, a fim de se estabelecer a conservação da espécie. Foram realizados cinco experimentos de enraizamento in vitro utilizando ANA, AIB, di e poliaminas, dithiothreitol e floroglucinol. As avaliações foram realizadas aos 30 e 60 dias quanto à porcentagem de enraizamento, número e comprimento de raiz. Para a introdução dos genótipos in vitro, foram utilizados segmentos nodais (1 cm) como explantes, contendo uma gema axilar ou apical, coletados de plantas mantidas em casa de vegetação, submetidos previamente à assepsia. As avaliações foram realizadas durante quatro semanas, quanto à porcentagem de contaminação dos explantes. Os resultados obtidos nas avaliações evidenciaram que a presença de compostos fenólicos no meio de cultura foi importante na promoção do enraizamento adventício in vitro de M. velutina e a metodologia de assepsia para a introdução de diferentes genótipos in vitro foi eficiente.

Mandevilla velutina (Apocynaceae) is a medicinal plant endemic to the Brazilian Cerrado, commonly known as "infalivel" and used by the population for treatments of inflammatory processes and accidents with snakes. This species is currently endangered due to extraction. The aims of this study were to optimize the protocol for in vitro rooting of M. velutina and to introduce different genotypes in the in vitro germplasm bank to establish the species conservation. Five experiments for in vitro rooting were conducted using NAA, IBA, di and polyamines, dithiothreitol and phloroglucinol. Evaluations were performed at 30 and 60 days as to rooting percentage, and root number and length. For the introduction of genotypes in vitro, nodal segments (1 cm) were used as explants; they had an axillary or apical bud and were collected from plants kept in a greenhouse after being subjected to asepsis. Evaluations were carried out for four weeks as to the percentage of explant contamination. Results showed that the presence of phenolic compounds in the culture medium was important to promote in vitro adventitious rooting in M. velutina and that the asepsis methodology for the introduction of in vitro of different genotypes was efficient.

Influência de diferentes meios de cultura sobre o crescimento de Pfaffia glomerata (Spreng. ) Pedersen (Amaranthaceae) para conservação in vitro / Influence of different culture media on the growth of Pfaffia glomerata (Spreng. ) Pendersen (Amaranthaceae) for conservation in vitro

Pfaffia glomerata ocorre em vários estados do Brasil e países limítrofes da região Sul às margens de rios e nas orlas das matas de galerias, é espécie hidrófita e heliófita. As raízes de espécies do gênero Pfaffia são usadas na medicina popular brasileira, especialmente como tônico, afrodisíaco e no controle do diabete. O objetivo deste trabalho foi estabelecer um banco de germoplasma in vitro de Pfaffia glomerata. O experimento em delineamento inteiramente casualizado foi conduzido com seis tratamentos: 1) MS + 2 por cento de sacarose + 4 por cento de sorbitol; 2) MS/2 + 2 por cento de sacarose + 4 por cento de sorbitol; 3) MS + 2 por cento de sacarose + 4 por cento de sorbitol + 2 mg L-1 de pantotenato de cálcio; 4) MS/2 + 2 por cento de sacarose + 4 por cento de sorbitol + 2 mg L-1de pantotenato de cálcio; 5) MS + 2 por cento de sacarose + 3 por cento de manitol + 2 mg L-1de pantotenato de cálcio; 6) MS/2 + 2 por cento de sacarose + 3 por cento de manitol + 2 mg L-1de pantotenato de cálcio. Os resultados obtidos foram submetidos à análise de variância e ao teste de separação de médias de Scott Knott. Os tratamentos um, três e quatro apresentaram, significativamente, o maior número de segmentos nodais por haste, quando comparados com os tratamentos dois, cinco e seis. O tratamento dois foi o mais indicado para a conservação in vitro da espécie por ter promovido menor crescimento das plantas (altura de 3,1±1,9 cm), alto índice de sobrevivência, 100 por cento de explantes com brotação e o maior número de brotos por explante, após seis meses de cultivo. Todas as plântulas produziram raízes e não houve formação de calos, também não ocorreu hiperhidricidade nos tratamentos avaliados. As plantas aclimatizadas apresentaram 100 por cento de sobrevivência no ambiente ex vitro. A manutenção de acessos de P. glomerata no banco de germoplasma in vitro é viável tanto do ponto de vista da conservação quanto economicamente.

Pfaffia glomerata occurs in several states of Brazil and its neighboring countries in the south region at riverbanks and gallery forests. It is a hydrophyte and heliophyte species. The roots of the genus Pfaffia are used in Brazilian folk medicine especially as tonic, aphrodisiac and to control diabetes. The aim of this work was to establish an in vitro germplasm bank for Pfaffia glomerata. The experiment was carried out in completely randomized design with six treatments: 1) DM + 2 percent sucrose + 4 percent sorbitol; 2) DM/2 + 2 percent sucrose + 4 percent sorbitol; 3) DM + 2 percent sucrose + 4 percent sorbitol + 2 mg L-1 calcium pantothenate; 4) DM/2 + 2 percent sucrose + 4 percent sorbitol + 2 mg L-1 calcium pantothenate; 5) DM + 2 percent sucrose + 3 percent mannitol + 2 mg L-1 calcium pantothenate; 6) DM/2 + 2 percent sucrose + 3 percent mannitol + 2 mg L-1 calcium pantothenate. Results were subjected to analysis of variance and Scott Knott test for mean grouping. Treatments 1, 3 and 4 had a significantly larger number of nodal segments per stem, compared to Treatments 2, 5 and 6. Treatment 2 was the most appropriate for the in vitro conservation of this species since it led to the lowest growth (3.1±1.9 cm height), high survival rate, 100 percent explants with sprouting, and the largest number of sprouts per explant after six months of culture. All seedlings produced root and showed no formation of calluses or hyperhydricidity under the evaluated treatments. Acclimatized plants showed 100 percent survival in the ex vitro environment. Maintaining P. glomerata accessions in an in vitro germplasm bank is viable both economically and for conservation.

Genetic variability among natural populations of Zaprionus indianus (Drosophilidae) in the States of São Paulo and Minas Gerais, Brazil.

Random amplified polymorphic DNA (RAPD) was used to detect polymorphisms among Zaprionus indianus fly populations collected from six municipalities in the States of São Paulo and Minas Gerais, Brazil. This species is an important, recently introduced fruit fly pest of figs and other fruit. Among 21 primers, 16 produced 73 reproducible polymorphic fragments; primer AM-9 produced the greatest number of polymorphic bands (nine), with 52% genetic variability among populations. Genetic divergence analysis of the Z. indianus populations demonstrated two major groups, named Western and Eastern groups. There was greater gene flow within than between groups. The correlation coefficient for genetic and geographic distances (Mantel test) was significant, demonstrating isolation by distance.

Genetic diversity of natural populations of Anemopaegma arvense (Bignoniaceae) in the Cerrado of São Paulo State, Brazil.

Catuaba (Anemopaegma arvense), a Bignoniaceae species endemic to Cerrado regions, shows anticancer properties and is widely used as a stimulant in traditional medicine. We evaluated the genetic diversity of seven populations found in the State of São Paulo, using random amplified polymorphic DNA markers. After optimization of the amplification reaction, 10 selected primers produced 70 reproducible bands, with 72.8% polymorphism. The greatest genetic diversity was observed within populations (71.72%). Variation estimates, theta(B) (0.2421) and Phi(ST) (0.283), obtained by inter- and intra-populational analysis of genetic variability of catuaba, indicated considerable population structure. However, the r value 0.346 (P = 0.099), calculated by the Mantel test, indicates that the genetic diversity among populations is not strongly structured in geographical space, although there appears to be a tendency towards structuring.

Assessment of genetic relationship between Klebsiella pneumoniae and Klebsiella oxytoca samples isolated from a dental office

The present study aimed to analyze the genetic similarity between genomic profiles of thirteen Klebsiella oxytoca and seven Klebsiella pneumoniae samples isolated from two different collections carried out in different places of dental offices. Random amplified polymorphic DNA (RAPD) technique and similarity coefficients (calculated by Sorensen-Dice and simple matching) were applied to determine their genetic profile of randomic DNA sequences. The majority of the isolates of K. pneumoniae and K. oxytoca presented similar coefficient values (¡Ý 0.80). Thus, it was possible to identify that strain dissemination occurred mainly via the hands of the surgeon-dentists and, finally, to determine the genetic similarity of the strains from dental office environments.(AU)

Direct organogenesis of Mandevilla illustris (Vell) Woodson and effects of its aqueous extract on the enzymatic and toxic activities of Crotalus durissus terrificus snake venom.

In order to produce explants of Mandevilla illustris (Vell) Woodson for the "Cerrado in vitro", the Germplasm Bank of UNAERP, we carried out a micropropagation protocol using MS or MS/3 medium supplemented with different concentrations of 6-benzyladeninepurine (BA), Zeatin or 2-isopentenyladenine for nodal segment growth, and alpha-naphthaleneacetic acid, indole-3-butyric acid (IBA) or 1,4 dithiothreitol for rooting. For nodal segments, all the cytokinins tested yielded similar results. However, 2.22 micro M BA is more economical to use. MS/3 medium supplemented with 0.49 micro M IBA was the most appropriate medium for rooting, resulting in 29% rooted explants. The crude aqueous extract from the subterranean system (SS) of M. illustris was assayed for its inhibitory action on the enzymatic activity of Crotalus durissus terrificus snake venom, isolated basic phospholipase A2 (CB) and crotoxin. It totally inhibited the phospholipase activity of crude Cdt venom and CB toxin and inhibited the phospholipase activity of crotoxin by 49%. The toxic action of both the crude venom and crotoxin was partially inhibited-there was a prolonged survival time and a 40.0% decrease in lethality.