Abstract Background: The association of scaffolds to repair extensive bone defects can contribute to their evolution and morphophysiological recomposition. The incorporation of particulate biomaterials into three-dimensional fibrin bioproducts together with photobiomodulation therapy (PBM) has potential and can improve regenerative medicine procedures. The objective of this experiment was to evaluate the effects of PBM therapy on critical size defects filled with xenogenic bone substitute associated with fibrin biopolymer. Methods: A critical defect of 8 mm was performed in 36 Wistar male adult rats that were divided into four groups. Groups BC and BC-PBM were defined as controls with defects filled by a clot (without or with PBM, respectively) and groups XS and XS-PBM that comprised those filled with biocomplex Bio-OssTM in association with fibrin biopolymer. PBM was applied immediately after the surgery and three times a week every other day, with the parameters: wavelength of 830 nm, energy density 6.2 J/cm2, output power 30 mW, beam area of 0.116 cm2, irradiance 0.258,62 W/cm2, energy/point 0.72 J, total energy 2.88 J. Fourteen and 42 days after the surgery, animals were euthanatized and subjected to microtomography, qualitative and quantitative histological analysis. Results: The BC-PBM and XS-PBM groups had a similar evolution in the tissue repair process, with a higher density of the volume of new formed bone in relation to the groups without PBM (p = 0.04086; p = 0.07093, respectively). Intense vascular proliferation and bone deposition around the biomaterial particles were observed in the animals of the groups in which biocomplex was applied (XS and XS-PBM). Conclusion: PBM therapy allowed an improvement in the formation of new bone, with a more organized deposition of collagen fibers in the defect area. Biocomplex favored the insertion and permanence of the particulate material in bone defects, creating a favorable microenvironment for accelerate repair process.

Abstract Squash is a food that meets the requirements of a healthy diet and is highly appreciated by consumers for its mild flavour and high nutritional value. In southern Brazil, many farmers grow several pumpkin landraces that are widely used in food. The consumption of its fruits and derivatives benefits human health due to the bioactive compounds, minerals, and antioxidants in the fruit pulp, making pumpkin a functional food. The objective of this study was to characterize the bioactive compounds, antioxidant activity, and minerals in the pulp of fruits of pumpkin landraces (Cucurbita maxima) from southern Brazil. Phenolic compounds, carotenoids, antioxidant activity, and minerals were evaluated in 10 accessions of pumpkin landraces from the Cucurbitaceae Genebank of Embrapa Temperate Agriculture (Pelotas, RS, Brazil). In mature fruits, the seeds and peels were discarded, and opposite longitudinal slices of pulp were manually prepared for analysis. All analyses were performed in triplicate. The data obtained showed genetic variability in total phenolic compounds, carotenoids, antioxidant activity, and minerals. Accessions C49, C307, and C216 had high levels of total phenolic compounds and carotenoids and high antioxidant activity. Of these accessions, accession C49 had the highest antioxidant capacity (504.1 μg·g-1). C216 and C178 were high in copper and iron, and the latter was highest in potassium, calcium, and phosphorus. Thus, accessions C216 and C178 have great potential for exploitation by genetic improvement programs for the development of biofortified cultivars in efforts to promote consumer health.

RESUMO O objetivo deste trabalho foi caracterizar morfologicamente variedades crioulas de Cucurbita maxima do sul do Brasil. Nove variedades crioulas de C. maxima foram caracterizadas com sete descritores quantitativos e 13 descritores qualitativos. As sementes foram cultivadas em casa de vegetação, em sacos de poliestireno preto, preenchidos com substrato. Quando as plantas apresentavam no mínimo duas folhas definitivas, 20 mudas de cada variedade foram transplantadas para o campo experimental, em delineamento completamente casualizado. Os dados quantitativos foram submetidos à análise de variância e foi utilizado o teste de Tukey para a comparação de médias, com o programa SAS 9.2. Os dados de caracteres quantitativos foram submetidos à análise de componentes principais, com o programa R. Para os caracteres qualitativos, a caracterização foi expressa pela moda de cada acesso, para cada descritor. Os caracteres qualitativos foram analisados como variáveis multicategóricas. A matriz de similaridade foi gerada no programa computacional GENES e foi construído um dendrograma, pelo método UPGMA, com o programa NTSYS. Ficou evidenciada a variabilidade genética para os caracteres quantitativos e qualitativos avaliados. A análise de variância evidenciou diferenças significativas entre as médias. Peso, formato, cor do fruto, espessura da casca e número de sementes por fruto são caracteres relevantes para a indicação de variedades crioulas promissoras para o melhoramento genético, com potencial para uso como fontes de genes e desenvolvimento de cultivares.

ABSTRACT The objective of this work was to morphologically characterize Cucurbita maxima landraces. Nine landraces were characterized by seven quantitative descriptors and 13 qualitative descriptors. The seeds were cultivated in black polystyrene bags filled with substrate in greenhouse. When the plants had at least two definitive leaves, 20 seedlings of each landrace were transplanted to the experimental field, spaced 2.5 m between plants and 4 m between rows, in a completely randomized design. Quantitative data were subjected to analysis of variance. Tukey test was used for the comparison of means, using the SAS 9.2 program. Data of quantitative traits were subjected to the principal component analysis, using the R program. For the qualitative traits, the characterization was expressed by the mode of each accession for each descriptor. Qualitative traits were analyzed as multicategorical variables. Similarity matrix was generated in the GENES program, and a dendrogram was constructed by the UPGMA method using the NTSYS program. Genetic variability was evidenced for the quantitative and qualitative traits evaluated. The analysis of variance showed significant differences between the means. Weight, shape, fruit color, bark thickness, and number of seeds per fruit are relevant characters to indicate promising landraces for breeding, with potential for use as sources of genes and development of cultivars.

Resumo: O objetivo deste trabalho foi avaliar a dinâmica evolutiva do pool genético silvestre-arvense-doméstico da abóbora (Cucurbita argyrosperma) ao estimar os níveis de diversidade genética e o fluxo gênico em sua provável área domesticação. Foram coletadas nove populações, tendo-se extraído o DNA de folhas jovens de cerca de 20 plantas de cada população. Os fragmentos de DNA foram amplificados por meio de oito pares de iniciadores de sequência simples repetida (SSR) e separados por eletroforese em 5% de gel de desnaturação de poliacrilamida. Estimaram-se a diversidade genética e o fluxo gênico para as populações, e utilizou-se o agrupamento bayesiano para determinar os níveis de infiltração gênica e probabilidade de ancestralidade. A investigação etnobotânica mostrou que a dinâmica evolutiva na área ocorreu em cinco cenários ecológicos diferentes. Foram identificados 87 alelos com 75 e 100% de loci polimórficos. A maior diversidade genética nas populações arvense-domésticas pode ter sido resultante da recombinação devido ao alto fluxo gênico entre estas populações, ocasionado pelos polinizadores e pela seleção humana. Há grande fluxo gênico entre as populações silvestres e cultivadas de C. argyrosperma no centro de domesticação, o que indica a importância da conservação e da manutenção destes recursos genéticos.

Abstract: The objective of this work was to evaluate the evolutionary dynamics of the wild-weedy-domestic gene pool of Cucurbita argyrosperma squash by estimating the levels of genetic diversity and gene flow in the putative area of its domestication. Nine populations were collected, and DNA was extracted from young leaves harvested separately from approximately 20 individuals in each population. The DNA fragments were amplified with eight pairs of SSR primers and separated by electrophoresis in 5% denaturing polyacrylamide gels. Genetic diversity and the amount of gene flow were estimated in the populations, and Bayesian grouping was used to determine the levels of gene infiltration and probability of ancestry. The ethnobotanical exploration indicated that the evolutionary dynamics in the area occurred under five different ecological scenarios. Eighty-seven alleles with 75% to 100% polymorphic loci were identified. The greater genetic diversity in the weedy-domestic populations may have been the product of recombination due to the high gene flow between these populations promoted by pollinators and human selection. There is high gene flow between the wild and cultivated populations of C. argyrosperma in its domestication centre, highlighting the importance of conserving and maintaining these genetic resources.

Abstract The objective of this work was to evaluate the genetic variability for the synthesis of bioactive compounds and minerals in pumpkin (Cucurbita moschata) landraces. Total phenolic compounds, carotenoids, antioxidant activity and minerals were evaluated in 10 accessions of Cucurbita moschata landraces from the Genebank of Embrapa Temperate Agriculture (Pelotas - RS, Brazil). Twenty plants of each accession were cultivated in the field during the spring/summer of 2013/2014. After harvesting of mature fruits, the seeds were discarded and opposite longitudinal portions of the pulp were manually prepared for analysis of the bioactive compounds. For the determination of minerals, pumpkin samples were frozen in plastic bags, and after freeze-dried and milled. All analysis were performed in triplicate. The data obtained showed high genetic variability for the synthesis of phenolic compounds, carotenoids, antioxidant activity and minerals. The accessions C52, C81, C267 e C389 showed high levels of antioxidants and minerals, being recommended for use in pumpkin breeding programs. The accessions C52 and C389 are promising, especially because they present the highest levels of total carotenoids.

Atualmente, muitas variedades crioulas de Capsicum baccatumsão cultivadas pelos agricultores no Brasil. Essas populações são fontes de genes para tolerância ou resistência aos estresses bióticos e abióticos. Porém, grande parte dessas variedades está sendo perdida devido à erosão genética causada pela substituição de cultivares. A fim de conservar e caracterizar as variedades crioulas, a Embrapa Clima Temperado mantém, desde 2002, um Banco Ativo de Germoplasma de Capsicum. Com o objetivo de caracterizar variedades crioulas de C. baccatum com marcadores moleculares de microssatélites, foram analisados vinte acessos que integram o acervo do Banco Ativo de Germoplasma de Capsicum da Embrapa Clima Temperado. O DNA foi extraído individualmente das folhas jovens de cinco plantas de cada acesso. Foram analisados oito loci de microssatélites, identificando um total de 43 alelos. Todos os loci analisados foram polimórficos. A análise molecular da variância mostrou uma diversidade genética de 51,37% entre acessos e 48,63% dentro dos acessos, o que evidenciou a existência de segregação em diversas variedades crioulas de pimentas avaliadas. Os grupos obtidos pela análise de agrupamento não tiveram relação com as características morfológicas dos frutos. A maior proporção da variabilidade genética encontra-se entre as diferentes variedades crioulas, embora também exista variabilidade genética dentro de cada acesso.

Currently, many landraces of C. baccatum are grown by farmers in Brazil. These populations are sources of genes for resistance or tolerance to abiotic and biotic stresses. However, most of these landraces are being lost due to genetic erosion caused by substitution of cultivars. In order to characterize and conserve landraces, since 2002 Embrapa Temperate Agriculture maintains an Active Germplasm Bank of Capsicum. This work has the objective to characterize landraces of C. baccatum using microsatellite markers. 20 accessions from the Active Germplasm Bank of Capsicum of Embrapa Temperate Agriculture were analyzed. DNA was extracted individually from young leaves collected from five plants of each accession. Eight microsatellite loci were analyzed. All loci were polymorphic. Molecular analysis of variance showed values of genetic diversity as 51.37% among accessions and 48.63% within the accessions. This fact showed the existence of segregation in many of these pepper landraces. The groups obtained by cluster analysis did not correlate with morphological fruit traits. Most of the genetic variability is found among different landraces, although there is also genetic variability within each accession.

No Sul do Brasil são cultivadas variedades locais das cinco espécies domesticadas de Cucurbita. Dentre elas, as menos estudadas são C. argyrosperma e C. ficifolia. Análises de microssatélites são importantes para a caracterização molecular dessas duas espécies, mas existe a necessidade de primers específicos, ainda não disponíveis. O objetivo desse trabalho foi avaliar a transferibilidade de primers e otimizar a amplificação de locos SSR de C. pepo em C. argyrosperma e C. ficifolia, bem como averiguar a diversidade genética entre diferentes acessos de germoplasma destas espécies. Foram utilizados dez acessos de C. pepo, nove de C. argyrosperma e cinco de C. ficifolia do Banco Ativo de Germoplasma de Cucurbitáceas da Embrapa Clima Temperado. O DNA foi extraído em bulk das folhas de cinco indivíduos de cada acesso. Foram usados 40 primers SSR desenvolvidos para C. pepo. Em C. pepo, 35 primers (87,5%) amplificaram, em C. argyrosperma, 34 primers (85%) e em C. ficifolia, 32 primers (80%). Portanto a taxa de transferibilidade de locos SSR de C. pepo para C. argyrosperma e C. ficifolia foi alta, indicando que podem ser empregados na caracterização molecular destas espécies.

Landraces of five domesticated species of Cucurbita are cultivated in southern Brazil. Among them C. argyrosperma and C. ficifolia are the least studied. Microsatellite markers are important for the molecular characterization of these two species, but there is a need for specific primers, not yet available. The aim of this study was to evaluate the transferability of primers and optimize the amplification of SSR loci in C. pepo, C. argyrosperma and C. ficifolia and also evaluate genetic diversity in different accessions of germplasm in these species. We used ten accessions of C. pepo, nine of C. argyrosperma and five of C. ficifolia from the Genebank of Cucurbitaceae from Embrapa Temperate Agriculture. DNA was extracted in bulk from the leaves of five individuals of each accession. 40 SSR primers developed for C. pepo were used. In C. pepo 35 primers (87.5%) amplified, in C. argyrosperma, 34 primers (85%), and in C. ficifolia, 32 primers (80%). The transferability of SSR loci in C. pepo to C. argyrosperma and C. ficifolia was 85 and 80% respectively. The transferability of SSR loci from C. pepo to C. argyrosperma and C. ficifolia was 85 and 80% respectively. Thus, the transferability rate of SSR loci developed for C. pepo to C. argyrosperma and C. ficifolia was high, indicating that they can be used in molecular characterization of these species.

Dentre as cinco espécies de abóboras domesticadas, Cucurbita pepo apresenta a maior variabilidade para características de fruto, como cor, formato e tamanho. O Banco Ativo de Germoplasma de Cucurbitáceas da Embrapa Clima Temperado conta com 61 acessos desta espécie, todos estes, variedades crioulas cultivadas no sul do Brasil. Estas variedades crioulas fazem parte da cultura regional, na gastronomia, no artesanato e na ornamentação de ambientes. O objetivo deste trabalho foi avaliar a variabilidade genética entre e dentro de variedades crioulas de C. pepo cultivadas no Rio Grande do Sul, utilizando marcadores microssatélites. Foram avaliados 10 acessos de variedades crioulas de C. pepo. que apresentavam diferentes formatos e tamanhos de fruto, cores e texturas da casca. O DNA genômico foi extraído individualmente de cinco plantas de cada acesso. Foram analisados 40 locos de microssatélites. Destes, 34 amplificaram, identificando 100 alelos (variação de um a cinco alelos por loco). Dos locos analisados 85,3% foram polimórficos, evidenciando a variabilidade genética entre os acessos. A análise molecular da variância mostrou que 45,39% da variabilidade genética é atribuída à variação dentro dos acessos e 54,60% a diferenças entre acessos. Assim, apesar de haver variabilidade genética dentro dos acessos, a maior proporção da variabilidade em C. pepo encontra-se distribuída entre as diferentes variedades crioulas. A variação dentro dos acessos é explicada pela alogamia da espécie, pelo intercâmbio de sementes das variedades crioulas entre os agricultores e também pelos mesmos cultivarem mais de uma variedade de Cucurbita na mesma área.

Among the five cultivated species of pumpkins, Cucurbita pepo has the highest variability for fruit characteristics, such as color, shape and size. The Active Germplasm Bank of Cucurbitaceae from Embrapa Clima Temperado (Brazil) has 61 accessions of this species, all of them landraces grown in southern Brazil. These landraces are part of regional culture, in gastronomy (preparation of traditional dishes) for crafts and ornaments. The aim of this study was to evaluate genetic variability within and among landraces of C. pepo cultivated in Rio Grande do Sul state, by microsatellite markers. Ten accessions of C. pepo landraces with different shapes and sizes of fruit, and colors and textures of fruit skin were evaluated. Genomic DNA was extracted individually from five plants of each accession. Forty microsatellite loci were analyzed, and 34 amplified. In total 100 alleles were found (one to five alleles per locus). From the analysed loci, 85.3% were polymorphic, showing the presence of genetic variability among the accessions. The analysis of molecular variance showed that 45.39% of genetic variability is attributed to differences within accessions, and 54.60% is attributable to variation among accessions. Thus, despite the presence of genetic variability within accessions, the largest proportion of variability in C. pepo is distributed among the accessions. The genetic variability found within accessions can be due to the allogamy, the exchange of landraces seeds between the farmers and also by cultivation of more than one Cucurbita landrace in the same area.

As pimentas do gênero Capsicum apresentam grande importância para o mercado de condimentos e para o uso ornamental no Brasil. A estimativa da diversidade genética é importante na escolha de progenitores de programa de melhoramento genético. Este trabalho teve por objetivo verificar a eficiência de descritores multicategóricos de flores para estimar a dissimilaridade genética entre acessos de Capsicum chinense, do Banco Ativo de Germoplasma Capsicum, da Embrapa Clima Temperado. O experimento foi realizado no período de agosto de 2009 a março de 2010, no campo experimental da Embrapa Clima Temperado, em blocos ao acaso, utilizando-se 22 acessos, com dez plantas por parcela. Para a caracterização morfológica das flores, foram avaliadas cinco flores de cada planta, de dez plantas por acesso. Foram utilizados 15 descritores, sendo cinco quantitativos e dez qualitativos multicategóricos. Foram realizadas análises de comparação de médias, utilizando-se agrupamento pelos métodos UPGMA e de Tocher, para os dados quantitativos, e agrupamento pelo método de Tocher, para os dados qualitativos. Os acessos estudados apresentam ampla diversidade genética em relação a descritores de flores, existindo grande variabilidade entre os acessos avaliados, o que recomenda seu uso em programas de melhoramento genético. O uso de descritores multicategóricos de flores é eficiente para estimar a dissimilaridade genética entre acessos de Capsicum chinense.

Capsicum peppers are extremely important in Brazil for the spice market and as ornamental. The estimation of genetic diversity is important for the choice of parents in a breeding program. This study aimed to evaluate the efficiency of multicategoric descriptors for flowers to estimate the genetic dissimilarity among Capsicum chinense accessions from the Capsicum Active Germplasm Bank of Embrapa Temperate Agriculture The experiment was conducted from October 2009 to March 2010 in the experimental field of Embrapa Temperate Agriculture, with 22 accessions evaluated in randomized blocks and ten plants per plot. Five flowers of each plant, ten plants per access were used for the morphological characterization of the flowers. Five quantitative and ten qualitative multicategoric descriptors were used, making a total of 15. Mean comparison tests were carried out, and the clustering methods UPGMA and Tocher were used for quantitative data, and the Tocher method for qualitative data. The accessions showed wide genetic diversity in relation to descriptors of flowers, with great variability among accessions, which recommends its use in breeding programs. The use of multicategoric descriptors for flowers is efficient to estimate the genetic dissimilarity among accessions of Capsicum chinense.