Genetic variation among sweet, purple, and yellow passion fruit accessions was assessed using inter-simple sequence repeat (ISSR) markers. Eighteen ISSR primers were used to evaluate 45 accessions. The number of polymorphic bands per primer varied from 4 to 22, with 12.4 bands per primer on average. Nei's genetic distance between accessions ranged from 0.04 to 0.35. Clustering using the neighbor-joining method resulted in the formation of 11 major clusters. It was not possible to classify the accessions according to their geographic origin, showing that there is no structure in the gene bank. The overall mean Shannon-Weaver diversity index was 0.32, indicating good resolution of genetic diversity in passion fruit germplasm using ISSR markers. Our results indicate that ISSR can be useful for genetic diversity studies, to provide practical information for parental selection and to assist breeding and conservation strategies.

Conventional methods for the selection of papaya pure lines are time-consuming. Procedures involving the use of molecular markers and the indirect selection of homozygous plants can reduce this time considerably. The objective of this study was to evaluate the informativeness of a microsatellite marker set when used in marker-assisted selection (MAS) for the development of new papaya lines. Eighty-three lines originating from two segregating F 3 populations and from papaya germplasm were used for the molecular analysis of 27 microsatellite primers. Twenty polymorphic microsatellite primers were identified, allowing the identification of 86 alleles, with an average of 3.18 alleles per primer. The observed heterozygosity values were low for both the markers (0.00-0.29) and the individual lines (0.00-0.35). The inbreeding coefficient (f) ranged from 0.634 to 1.00. Eleven lines with f = 1.00 and 18 lines with f varying from 0.953 to 0.961 were identified. In addition, papaya lines showed high genetic diversity, which will certainly contribute to the development of new varieties. Our results show that the use of microsatellites in MAS is a quick and effective procedure for the development of papaya lines.

-Due to the low genetic variability reported in the commercial plantations of papaya (Carica papaya L.), the objective of this study was analyze the genetic diversity of 32 genotypes including cultivars, landraces, inbred lines, and improved germplasm using the AFLP technique (Amplified Fragment Length Polymorphism). The genetic distance matrix was obtained using the Nei and Li genetic distance and clustering was performed using the unweighted pair-method with arithmetic mean (UPGMA). Using 11 combinations of EcoRI/MseI primers, 383 polymorphic bands were obtained. On average, 34.8 polymorphic bands were obtained per primer combination. Five clusters were formed. The traditional cultivar 'Sunrise' and the inbred line CMF-L30-08 were the closest genotypes, and the improved germplasm (CMF041) and landrace (CMF233) the most distant. The main papaya cultivars commercially grown in Brazil, as well as four inbred lines and three improved germplasm, were clustered together, however, were not grouped in the same branch. The genetic distance between the Sunrise and Golden cultivars was 0.329, and even arising from mutation and selection within the Sunrise variety, the Golden stores considerable genetic variability. Additional variability was observed in the inbred lines derived from papaya breeding program at Embrapa Cassava and Fruits. Index terms: papaya breeding, genetic resources, genetic diversity, Carica papaya L.. CARACTERIZAÇÃO MOLECULAR DE GENÓTIPOS DE MAMOEIRO COM USO DE MARCADORES DO TIPO AFLPRESUMO -Em virtude da baixa variabilidade genética relatada nos plantios comerciais de mamoeiro (Carica papaya L.), o objetivo deste trabalho foi analisar a divergência genética de 32 genótipos incluindo cultivares, variedades locais, linhagens e germoplasma melhorado, utilizando a técnica de AFLP (Amplified Fragment Length Polymorphism). A matriz de distâncias genéticas foi obtida com a distância de Nei e Li, e o agrupamento foi realizado utilizando o método UPGMA (unweighted pair-method with arithmetic mean). Utilizaram-se 11 combinações de iniciadores com corte das enzimas EcoRI / MseI. Foram geradas 383 bandas polimórficas, com média de 34,8 por combinação de iniciadores. O agrupamento dos dados permitiu a formação de cinco grupos, sendo a cultivar Sunrise e a linhagem CMF-L30-08 os genótipos mais relacionados, e CMF041 (germoplasma melhorado) e CMF233 (variedade local), os mais dissimilares. As cultivares de mamoeiro mais cultivadas no Brasil, bem como quatro linhagens e três acessos de germoplasma melhorado foram agrupadas, porém não no mesmo ramo. Mesmo sendo oriunda de mutação e seleção dentro de Sunrise, a cultivar Golden ainda possui considerável variabilidade genética, uma vez que a distância genética entre estas cultivares foi de 0,329. Variabilidade adicional foi observada nas linhagens do programa de melhoramento. Termos para indexação: melhoramento, recursos genéticos, diversidade genética, Carica papaya L..Rev. Bras.

Ayahuasca is a hallucinogenic beverage produced from the decoction of Banisteriopsis caapi (Bc) and Psychotria viridis (Pv), β-carboline- and N,N-dimethyltryptamine(DMT)-containing plants, respectively. Accumulating evidence suggests that ayahuasca may have therapeutic effects on ethanol abuse. It is not known, however, whether its effects are dependent on the presence of DMT or if non-DMT-containing components would have therapeutic effects. The aim of the present study was to investigate the rewarding properties of ayahuasca (30, 100, and 300 mg/kg, orally), Bc (132, 440, and 1320 mg/kg, orally) and Pv (3.75, 12.5 and 37.5 mg/kg, i.p.) extracts and their effects on ethanol (1.8 g/kg, i.p.) reward using the conditioned place preference (CPP) paradigm in male mice. Animals were conditioned with ayahuasca, Bc or Pv extracts during 8 sessions. An intermediate, but not a high, dose of ayahuasca induced CPP in mice. Bc and Pv did not induce CPP. Subsequently, the effects of those extracts were tested on the development of ethanol-induced CPP. Ayahuasca, Bc or Pv were administered before ethanol injections during conditioning sessions. While Bc and Pv exerted no effects on ethanol-induced CPP, pretreatment with ayahuasca blocked the development of CPP to ethanol. Finally, the effects of a post-ethanol-conditioning treatment with ayahuasca, Bc or Pv on the expression of ethanol-induced CPP were tested. Animals were conditioned with ethanol, and subsequently treated with either ayahuasca, Bc or Pv in the CPP environment previously associated with saline or ethanol for 6 days. Animals were then reexposed to ethanol and ethanol-induced CPP was quantified on the following day. Treatment with all compounds in the ethanol-paired environment blocked the expression of ethanol-induced CPP. Administration of an intermediate, but not a high, dose of ayahuasca and Bc, as well as Pv administration, in the saline-paired compartment blocked the expression of ethanol-induced CPP. The present study sheds light into the components underlying the therapeutic effects of ayahuasca on ethanol abuse, indicating that ayahuasca and its plant components can decrease ethanol reward at doses that do not exert abuse liability. Importantly, the treatment environment seems to influence the therapeutic effects of ayahuasca and Bc, providing important insights into clinical practice.

The objective of this work was to characterize 203 lineages and five parents of Ricinus communis L. from the germplasm bank at the Federal University of Recôncavo da Bahia (UFRB), which was established by the Genetic Improvement and Biotechnology Program (NBIO) at the Center for Agrarian, Environmental and Biological Sciences. The study used 35 morpho-agronomic descriptors, proposed by the Ministry of Agriculture, Livestock and Supply, and 12 quantitative descriptors suggested by NBIO. The experiment was conducted in a randomized block design, composed of four blocks, in the experimental field at UFRB in 2014. The frequency and entropy level of the qualitative descriptors were estimated with the Renyi procedure, and an analysis of variance was used for the quantitative descriptors. The analyses were made with the statistical program R. Of the qualitative morpho-agronomic descriptors evaluated, 22.86% had a high level of entropy (above 1.0), and all 12 quantitative descriptors showed significant differences. This indicates genetic variability in the germplasm bank and a satisfactory performance for most of the descriptors evaluated, as well as the possibility of direct and indirect use of the lineages and parents in genetic improvement programs of the species.

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The cupuassu tree (Theobroma grandiflorum) (Willd. ex Spreng.) Schum. is a fruitful species from the Amazon with great economical potential, due to the multiple uses of its fruit´s pulp and seeds in the food and cosmetic industries, including the production of cupulate, an alternative to chocolate. In order to support the cupuassu breeding program and to select plants presenting both pulp/seed quality and fungal disease resistance, SSRs from Next Generation Sequencing ESTs were obtained and used in diversity analysis. From 8,330 ESTs, 1,517 contained one or more SSRs (1,899 SSRs identified). The most abundant motifs identified in the EST-SSRs were hepta- and trinucleotides, and they were found with a minimum and maximum of 2 and 19 repeats, respectively. From the 1,517 ESTs containing SSRs, 70 ESTs were selected based on their functional annotation, focusing on pulp and seed quality, as well as resistance to pathogens. The 70 ESTs selected contained 77 SSRs, and among which, 11 were polymorphic in cupuassu genotypes. These EST-SSRs were able to discriminate the cupuassu genotype in relation to resistance/susceptibility to witches’ broom disease, as well as to pulp quality (SST/ATT values). Finally, we showed that these markers were transferable to cacao genotypes, and that genome availability might be used as a predictive tool for polymorphism detection and primer design useful for both Theobroma species. To our knowledge, this is the first report involving EST-SSRs from cupuassu and is also a pioneer in the analysis of marker transferability from cupuassu to cacao. Moreover, these markers might contribute to develop or saturate the cupuassu and cacao genetic maps, respectively.

Cupuassu (Theobroma grandiflorum [Willd. ex Spreng.] Schum) is a species of high economic importance in Brazil with great potential at international level due to the multiple uses of both its seeds and pulp in the industry of sweets and cosmetics. For this reason, the cupuassu breeding program focused on the selection of genotypes with high pulp and seed quality—selection associated with the understanding of the mechanisms involved in fruit formation. Gene expression is one of the most used approaches related to such understanding. In this sense, quantitative real-time PCR (qPCR) is a powerful tool, since it rapidly and reliably quantifies gene expression levels across different experimental conditions. The analysis by qPCR and the correct interpretation of data depend on signal normalization using reference genes, i.e. genes presenting a uniform pattern of expression in the analyzed samples. Here, we selected and analyzed the expression of five genes from cupuassu (ACP, ACT, GAPDH, MDH, TUB) to be used as candidates for reference genes on pulp and seed of young, maturing and mature cupuassu fruits. The evaluation of the gene expression stability was obtained using the NormFinder, geNorm and BestKeeper programs. In general, our results indicated that the GAPDH and MDH genes constituted the best combination as reference genes to analyze the expression of cupuassu samples. To our knowledge, this is the first report of reference gene definition in cupuassu, and these results will support subsequent analysis related to gene expression studies in cupuassu plants subjected to different biotic or abiotic conditions as well as serve as a tool for diversity analysis based on pulp and seed quality.