Modern genotyping techniques, such as SNP analysis and genotyping by sequencing (GBS), are hampered by poor DNA quality and purity, particularly in challenging plant species, rich in secondary metabolites. We therefore investigated the utility of a pre-wash step using a buffered sorbitol solution, prior to DNA extraction using a high salt CTAB extraction protocol, in a high throughput or miniprep setting. This pre-wash appears to remove interfering metabolites, such as polyphenols and polysaccharides, from tissue macerates. We also investigated the adaptability of the sorbitol pre-wash for RNA extraction using a lithium chloride-based protocol. The method was successfully applied to a variety of tissues, including leaf, cambium and fruit of diverse plant species including annual crops, forest and fruit trees, herbarium leaf material and lyophilized fungal mycelium. We consistently obtained good yields of high purity DNA or RNA in all species tested. The protocol has been validated for thousands of DNA samples by generating high data quality in dense SNP arrays. DNA extracted from Eucalyptus spp. leaf and cambium as well as mycelium from Trichoderma spp. was readily digested with restriction enzymes and performed consistently in AFLP assays. Scaled-up DNA extractions were also suitable for long read sequencing. Successful RNA quality control and good RNA-Seq data for Eucalyptus and cashew confirms the effectiveness of the sorbitol buffer pre-wash for high quality RNA extraction.

In this paper random amplified polymorphic DNA (RAPD) was used to evaluate the degree of among-population differentiation and associated spatial patterns of genetic divergence for Dipteryx alata Vogel populations from Cerrado region of central Brazil, furnishing support for future programs of conservation of this species. We analyzed patterns of genetic and spatial population structure using 45 RAPD loci scored for 309 trees, sampled from five different regions with two populations each. Genetic structure analysis suggested that panmixia null hypothesis can be rejected, with significant among-population components of 15%. Hierarchical partition by Analysis of Molecular Variance (AMOVA) shows that 5% of genetic variation is within regions, whereas 10% of variation is among regions, and these results were confirmed by a Bayesian analyses on HICKORY. The Mantel correlogram revealed that this divergence is spatially structured, so that local populations situated at short geographic distances could not be considered independent units for conservation and management. However, genetic discontinuities among populations were found in the northwest and southeast parts of the study area, corresponding to regions of recent socio-economic expansion and high population density, respectively. Taking both geographic distances and genetic discontinuities into account it is possible to establish a group of population to be conserved, covering most of D. alata geographic distribution and congruent with previously established priority areas for conservation in the Cerrado region.

• Premise of the study: Microsatellite markers were developed for the population genetic analyses of the neotropical tree Dipteryx alata (Fabaceae). • Methods and Results: Microsatellites were developed from a genomic shotgun library. Polymorphism at each microsatellite loci was analyzed based on 94 individuals from three populations. Eight loci amplified successfully and presented one to 10 alleles, and expected heterozygosities ranged from 0.097 to 0.862. Four loci also amplified in Pterodon emarginatus and presented similar polymorphism. • Conclusion: The eight microsatellite primer pairs are potentially suitable for population genetic studies and successfully amplified in another Fabaceae species.

We used microsatellite loci to test the paternity of two male jaguars involved in an infanticide event recorded during a long-term monitoring program of this species. Seven microsatellite primers originally developed for domestic cats and previously selected for Panthera onca were used. In order to deal with uncertainty in the mother's genotypes for some of the loci, 10000 values of W were derived by simulation procedures. The male that killed the two cubs was assigned as the true sire. Although the reasons for this behavior remain obscure, it shows, in principle, a low recognition of paternity and kinship in the species. Since the two cubs were not very young, one possibility is that the adult male did not recognize the cubs and killed them for simple territorial reasons. Thus, ecological stress in this local population becomes a very plausible explanation for this infanticide, without further sociobiological implications.

Resumo -O objetivo deste trabalho foi avaliar o padrão espacial da variabilidade genética entre plantas, dentro de três populações naturais de barueiro (Dipteryx alata Vogel), pela genotipagem por RAPD e técnicas de autocorrelação espacial. Os cinco iniciadores RAPD permitiram a codificação de 45 locos, utilizados nas análises de diversidade, estrutura e distribuição espacial da variabilidade genética entre populações. As populações apresentaram diversidade genética (Hs) com valor médio 0,314. Verificou-se que 12% da variação total se encontra entre as populações, o que indica que estas mantêm um considerável nível de variabilidade genética. Foi observada tendência de autocorrelação espacial positiva nas primeiras classes de distâncias, nas três populações, o que indica a formação de grupos de vizinhança com estruturação familiar, dentro das populações de barueiro. Entretanto, o tamanho desses grupos de vizinhança varia entre as populações; isso mostra que outros processos ecológicos influenciaram a distribuição espacial da variabilidade genética. As populações naturais de barueiro apresentam consideráveis níveis de diversidade genética, com base nos 45 locos RAPD avaliados.Termos para indexação: autocorrelação espacial, baru, conservação, estrutura familiar, fluxo gênico, RAPD. Spatial distribution of intrapopulational genetic variability in Dipteryx alataAbstract -The aim of this paper was to evaluate the spatial pattern of the genetic variability among plants, within local populations of "barueiro" (Dipteryx alata Vogel), using RAPD markers and spatial autocorrelation analyses. The five RAPD primers used allowed the coding of 45 loci used for standard analyses of diversity, structure and spatial distribution of genetic variability within each local population. These populations showed considerable amount of genetic diversity, with an average Hs of 0.314. From the total genetic variation 12% occur among the three local populations studied, indicating that each of these contains a significant amount of variation. Within each population, this variability is structured with significant and positive spatial autocorrelation in the first three distance classes, suggesting the existence of neighbor groups of genetically similar individuals. The size of these neighbor groups, however, varies among local populations; this shows that other ecological components affected the spatial structure of genetic diversity. Natural populations of "barueiro" present considerable levels of genetic diversity, based on 45 RAPD loci analysed.