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The Rhipicephalus (Boophilus) microplus tick is a major concern for the livestock market worldwide, as it causes serious economic damage. Plant-derived acaricides are an attractive alternative to control this ectoparasite and limit the development of resistance. Therefore, the aim of this study was to evaluate the acaricidal activity of Furcraea foetida leaf extract against engorged female R. (B.) microplus ticks. Our in vitro bioassays showed that the crude extract of leaves from F. foetida caused hemorrhagic swelling and skin lesions in the ticks, and three days of treatment caused 100% mortality. Dose-response assay indicated that this toxicity effect was dose-dependent. Similar effects were observed when the crude extract from F. foetida leaves was denatured by boiling at 100°C. These results suggest that the toxicity of the leaf extract might be associated with thermostable biomolecules. Together, our results show for the first time that the crude extract of F. foetida leaves has acaricidal activity against engorged female R. (B.) microplus ticks and it acts in a dose-dependent manner.

Advances in physiology and other fields are strongly associated with a solid base knowledge of biochemistry and cell metabolism. On the other hand, the complex and abstract nature of metabolic pathways, the traditional lecture method, and other factors made the teaching-learning process of biochemistry a challenging endeavor. To overcome this, we developed and tested a novel active learning tool called Interactive Metabolism ( iM-tool). The iM-tool was developed with simple and low-cost materials. We used it for interactive teaching of several metabolic pathways and physiological mechanisms for students enrolled in the Biochemistry subject belonging to different undergraduate courses. The results of evaluation tests showed that the iM-tool significantly (ANOVA, P < 0.01) and consistently improved the biochemistry knowledge of students in classrooms with up to 50 students for 7 different and consecutive academic semesters. A survey intended to mine students’ opinions on the tools showed significant satisfaction with the teaching using the iM-tool over traditional lecture-based teaching, and the iM-tool contributed to collaborative learning among students. Therefore, our results showed that the iM-tool improves the biochemistry and cell metabolism teaching-learning process in a more attractive and interactive manner.

Molecular markers have revolutionized analyses in population genetics, enabling precise estimates of the amount of genetic variability and how it is distributed within and among populations. The high diversity of bamboos, distributed throughout the world and of high economic relevance, has deserved several studies on molecular characterization. This chapter describes how distinct categories of molecular markers, such as isozymes, RAPD, AFLP, microsatellites, and SNP, have enabled the analysis of population genetic processes, assessments of the genetic diversity, and structure of natural populations and selected cultivars of bamboo species. One important application is their power of phylogenetic inference, enabling the distinction of the diverse set of bamboo species. With the genomic technologies, gene families have been characterized, mainly for Phyllostachys edulis, which has its genome sequenced and deposited to databases, enabling the detection of markers related with environmental constraints. As vegetative propagation is a common mechanism in bamboos and their cultivation relies on this strategy, molecular markers have been important for attesting genetic fidelity to their original source of propagules. Altogether, we provide a panorama of several applications of molecular markers to bamboo conservation and breeding.

Bamboos comprise more than 1600 species distributed throughout the world, according to recent updates. They are used for a wide range of applications such as utensils, charcoal, cellulose extraction, and hybrid microfibers. Bamboos also have an important ecological role in their environment of origin. Therefore, it is critical to recognize, catalogue, and investigate the genetic resources available for the distinct species. Establishing traits of economic value and their heritability is an important step toward breeding programs. Due to the limited sexual reproduction and their natural ability for vegetative propagation, several techniques have been developed for the propagation of genotypes of interest. We provide a brief panorama of methods used for clonal propagation of bamboo species, such as culm-cuttings and culm-layering. In vitro strategies that have been optimized for bamboo propagation is also described. Finally, we outline the significance and application of molecular markers into the management of bamboos. As components of natural communities and as sustainable alternatives for large-scale economic applications, with the advantage of fast growth and high biomass production, conservation and breeding of bamboos certainly must be given attention for future endeavors.

SUMMARY: Few studies have focused on the in vitro multiplication methods for Eucalyptus cloeziana. In this work we developed an indirect organogenesis protocol to obtain micropropagated plants. The interactions of plant growth regulators in juvenile tissues were evaluated. Hypocotyl, cotyledon and root tissues obtained from in vitro germinated seedlings were subjected to combinations of α-naphthaleneacetic acid (NAA), thidiazuron (TDZ) and 2,4-dichlorophenoxyacetic acid (2,4-D). In vitro callogenesis, adventitious bud induction, shoot elongation, and ex vitro survival and rooting were evaluated. Callogenic structures were observed in all evaluated tissues. Morphogenic characteristics related to the meristematic center was observed. The tissues that presented the callus mass were subcultured in a regeneration culture medium supplemented with 1.0 mg L-1 of 6-benzylaminopurine (BAP). Only hypocotyl and cotyledon induced adventitious bud and shoot elongation. Hypocotyl subjected to 2.0 mg L-1 TDZ had the highest number of elongated shoots. The two tissues (i.e., hypocotyl and cotyledon) combined with thidiazuron were characterized by the ex vitro survival of microcuttings and by adventitious rooting. Results confirmed tissue culture efficiency for microplants production by indirect organogenesis from hypocotyl and cotyledon cultivated with TDZ, and its implementation can be an alternative for forest tree breeding programs of E. cloeziana.

Bamboo species are an alternative for the composition of forest plantations. However, their potential has not been explored due to the hard time in producing large-scale clonal plants. Thus, the aim this work was to evaluate the in vitro establishment, bud multiplication and ex vitro rooting of Bambusa vulgaris. The first experiment tested different systemic and contact fungicide solutions, based on exposure time, during the establishment phase. Established explants were subjected to evaluation of residual fungicide effect on subcultures during the multiplication and elongation phases. The second experiment evaluated the influence of activated carbon on ex vitro survival and on adventitious rooting. Explant immersion in liquid culture medium added with 1.0 mL of fungicide for 120 hours has favored the in vitro establishment and reduced fungal contamination. On the other hand, it favored the shoot emission of shoots per explant during the multiplication phase. Both rooting induction culture medium and mini-incubator system use were effective in enabling adventitious root formation. The presence of activated carbon in the rooting induction culture medium resulted in a higher clonal plant survival rate. As espécies de bambus são uma alternativa para a composição de plantios florestais. Entretanto, esse potencial não tem sido explorado devido à dificuldade de produção de mudas clonais em larga escala. Assim, objetivo deste trabalho foi avaliar o estabelecimento in vitro, a multiplicação e o enraizamento ex vitro de Bambusa vulgaris. No primeiro experimento foram testadas diferentes soluções de fungicida sistêmico e de contato em relação ao tempo de exposição durante a fase de estabelecimento. Os explantes estabelecidos foram avaliados quanto ao efeito residual do fungicida durante subcultivos nas fases de multiplicação e alongamento. No segundo experimento, foi avaliada a influência do carvão ativado sobre a sobrevivência e enraizamento ex vitro. Durante o estabelecimento in vitro, a imersão de explantes em meio de cultura líquido contendo alíquota de 1,0 mL de fungicida durante 120 horas favoreceu o estabelecimento e reduziu a contaminação fúngica, enquanto na fase de multiplicação, houve o favorecimento da emissão de brotos por explante. O meio de cultura de indução ao enraizamento e uso de sistema de mini-estufim foram efetivos para a formação de raízes adventícias e a presença de carvão ativado resultou em uma maior sobrevivência das mudas clonais.

Swietenia macrophylla yields high-quality wood; however, its vulnerability to extinction coupled with challenges for its cultivation have brought attention to its replacement for alternative species such as the Khaya anthotheca. The species has been recently introduced to South America, with potential for the production of high-quality wood. However, limited background on breeding and efficient strategies for its vegetative propagation exist. Here, we achieved significant results with the application of cleft grafting and air layering for the propagation of K. anthotheca plants grown from seeds. First, we analyzed the compatibility of scions and rootstocks for intraspecific and interspecific cleft grafting combinations of K. anthotheca and S. macrophylla. Second, air layering was performed in K. anthotheca seedlings irrigated with three nutrient solution (100%, 50% and 25% of the initial concentration of nutrients) combined with the application of indole-3-butyric acid (IBA) to evaluate adventitious rooting. From cleft grafting, we achieved an overall graft compatibility and survival of 48% after 200 days. However, the interspecific combination of S. macrophylla (scion) and K. anthotheca (rootstock) implicated in no compatibility, while the reciprocal resulted in 52% of compatibility. Through air layering, the irrigation with the nutrient solution with at least 50% of the nutrients concentration and with IBA (3.0 or 8.0 g.L-1) resulted in the best adventitious rooting. Overall, we recommend cleft grafting, except for the combination S. macrophylla(scion) and K. anthotheca (rootstock), with no compatibility. Air layering might also be useful for the propagation of K. anthotheca genotypes in breeding programs.

Cochlospermum regium roots are used in popular medicine and its extract has diverse phytochemical molecules some with antimicrobial activity, consequently exposing this specie to genetic erosion risks. Thus, the objective of this study was to develop an in vitro multiplication protocol using chemical sterilization of culture medium. Therefore, explants obtained from apical buds of C. regium seedlings were inoculated into with 0.05mg L-1 NAA and 1mg L-1 BAP sterilized by chemical agent sodium hypochlorite (NaOCl) at 0.001%, 0.003% and 0.005% of active chlorine (Cl). Autoclaved culture medium was used as control. Result showed that the contamination by bacterial at 91 days of cultivation was significantly (P

A viticultura e geradora de renda tanto em nivel mundial como nacional. Entretanto, todas as regioes produtoras no Brasil sao afetadas pelo mildio (Plasmopara viticola), doenca que pode causar serios danos a cultura. O objetivo do trabalho foi avaliar a resistencia ao mildio de cultivares de uva introduzidas no norte fluminense. Para tanto foram avaliadas onze cultivares a saber, ?Rosalinda?; ?Italia?; ?Patricia?; ?Kyoho?; ?Red Globe?; ?Roberta?; ?Rubi?; ?Niagara Rosada?; ?Isabel?; ?Moscatel de Hamburgo? e ?Romana?. As plantas foram conduzidas em sistema de ?latada?, no espacamento 4,0 x 3,0 m, em delineamento experimental de blocos ao acaso, com tres repeticoes e quatro plantas por parcela. Foram realizadas duas avaliacoes quanto ao nivel da doenca, uma referente a leitura dos sintomas, por tres avaliadores que utilizaram uma escala de notas variando de 1 (resistente) a 5 (suscetivel), e a segunda seguindo a escala proposta por Azevedo (1997). Foi feita analise de variância e teste de comparacao de medias de Scott-Knott (P

O fator de alcalinização rápida ou RALF (do inglês, Rapid ALkalinization Factor) é um peptídeo hormonal ubíquo que induz a atividade de uma MAP quinase, bem como um aumento rápido do pH extracelular do meio de cultura de células em suspensão. O AtRALF1, uma das 37 isoformas de RALF presentes em Arabidopsis thaliana, é um peptídeo secretado de 5 kDa, que inibe o crescimento radicular e causa uma rápida mobilização de Ca2+ extra e intracelular. Em células eucarióticas, calmodulinas são proteínas bem caracterizadas por mediar a transdução de sinais de Ca2+ intracelular. Entretanto, em várias espécies vegetais incluindo Arabidopsis, também existem relatos de calmodulinas secretadas com funções ainda desconhecidas. Neste trabalho, foi caracterizada a AtCML38, uma proteína de Arabidopsis semelhante a calmodulina e que interage com o peptídeo AtRALF1. Análises in silico e por RT-PCR semi-quantitativo mostram a co-expressão de ambos os genes AtRALF1 e AtCML38 em raízes de plântulas de Arabidopsis. Apesar da ausência de um peptídeo sinal típico na proteína AtCML38, a localização sub-celular demonstra que esta é secretada através da via secretória RE-Golgi. O uso de uma versão truncada da AtCML38, cujos 87 primeiros aminoácidos foram retirados, indicou que o N-terminal é essencial para o seu direcionamento. Por meio de ensaios de complementação fluorescente bimolecular, foi demonstrado que AtCML38 interage com AtRALF1 no apoplasto de folhas de tabaco. Ensaios de pulldown indicam que esta interação é específica e dependente de Ca2+ e do pH. Um mutante por inserção de T-DNA, que não produz a proteína AtCML38 (cml38), mostrou-se insensível ao peptídeo AtRALF1 aplicado exogenamente, e suas raízes cresceram normalmente. Plantas transgênicas de Arabidopsis super-expressando o gene AtRALF1 (35S:AtRALF1) têm um fenótipo semi-anão com pronunciada redução do crescimento radicular. Contudo, quando plantas 35S:AtRALF1 foram cruzadas com o mutante cml38, suas progênies exibiram fenótipo e crescimento radicular normais, apesar do acúmulo do peptídeo AtRALF1. Juntos, os resultados mostram que AtCML38 interage com o peptídeo hormonal AtRALF1, e que a proteína AtCML38 é essencial para inibição do crescimento radicular causado pelo peptídeo. Rapid alkalinization factor (RALF) is a ubiquitous peptide hormone that induces a MAP kinase and a rapid increase in the pH of the extracellular media of cell suspension cultures. AtRALF1, one of the 37 RALF isoforms of Arabidopsis thaliana, is a secreted peptide of 5 kDa that inhibits root growth and causes a rapid mobilization of extra and intracellular Ca2+. In eukaryotic cells, calmodulin proteins are well-characterized to mediate intracellular Ca2+ signal transduction. Nevertheless, in several plant species including Arabidopsis, there are also reports of calmodulins being secreted with still unknown function. In this work, we characterized the AtCML38, a calmodulin-like protein from Arabidopsis as an AtRALF1-interacting protein. Analyses in silico and semiquantitative RT-PCR show co-expression of both AtCML38 and AtRALF1 genes in roots of Arabidopsis seedlings. Despite the fact that AtCML38 lacks a typical signal peptide, subcellular localization demonstrates that AtCML38 is secreted via the ER-Golgi secretory pathway. A truncated AtCML38, without the first 87 amino acids indicates that the N-terminal is essential for targeting. Through bimolecular fluorescence complementation assays, we showed that AtCML38 protein interacts with AtRALF1 in the apoplast of epidermal cells from tobacco leaves. Pull down assays indicate that this interaction is specific, Ca2+- and pH-dependent. A T-DNA insertion mutant defective in the AtCML38 protein (cml38) was insensitive to exogenously applied AtRALF1 peptide and their roots grow just as the wild type plants. Transgenic Arabidopsis plants overexpressing the AtRALF1 gene (35S:AtRALF1) have a semi-dwarf phenotype with a pronounced reduction in the root growth. However, when 35S:AtRALF1 plants are crossed with the mutant cml38, their progenies are normal looking and exhibit normal root growth in spite of the high accumulation of AtRALF1 peptide. Taken together, the results show that AtCML38 interacts with the peptide hormone AtRALF1 and that the protein AtCML38 is essential for the root growth inhibition caused by the peptide.

A viticultura é geradora de renda tanto em nível mundial como nacional. Entretanto, todas as regiões produtoras no Brasil são afetadas pelo míldio (Plasmopara viticola), doença que pode causar sérios danos a cultura. O objetivo do trabalho foi avaliar a resistência ao míldio de cultivares de uva introduzidas no norte fluminense. Para tanto foram avaliadas onze cultivares a saber, ?Rosalinda?; ?Itália?; ?Patrícia?; ?Kyoho?; ?Red Globe?; ?Roberta?; ?Rubi?; ?Niágara Rosada?; ?Isabel?; ?Moscatel de Hamburgo? e ?Romana?. As plantas foram conduzidas em sistema de ?latada?, no espaçamento 4,0 x 3,0 m, em delineamento experimental de blocos ao acaso, com três repetições e quatro plantas por parcela. Foram realizadas duas avaliações quanto ao nível da doença, uma referente a leitura dos sintomas, por três avaliadores que utilizaram uma escala de notas variando de 1 (resistente) a 5 (suscetível), e a segunda seguindo a escala proposta por Azevedo (1997). Foi feita análise de variância e teste de comparação de médias de Scott-Knott (P