Optimized method for dna extraction and PCR amplification in aroeira tree

Molecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of ß-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers.

Medicinal plants of the brazilian cerrado: knowing to preserve / Plantas medicinais do cerrado brasileiro: sabendo preservar

The Cerrado is the second largest biome in Brazil and presents an immeasurable and still underexplored ecological diversity. Despite the exuberance of its endemic species, it is one of the 25 global hotspots, due to a high natural biodiversity wealth along with an expressive environment destruction. In this study, we surveyed the knowledge on medicinal use of Cerrado plants held by individuals living in a predominantly Cerrado region. A semi-structured individual questionnaire for qualitative socio-economic characterization and medicinal use of plants was applied to Public Health Service users of the city of Assis, State of Sao Paulo, Brazil. This study was approved by the Ethics Committee of the Assis Regional Hospital (Protocol 4812010). Out of 149 respondents, 90.6% reported knowledge and/or use of plants for therapeutic purposes. Among 115 mentioned plant species, only 11.4% belong to the Cerrado. We also evaluated how matching were the reported uses and the respective published data from scientific studies on the plants' medicinal properties. It was verified that for the few Cerrado plants cited, the respondents described several medicinal applications not scientifically described. More precisely, 60.5% of all medicinal applications were not found in the scientific literature. Besides that, many of the therapeutic effects described in the literature for the Cerrado species were not cited by the interviewed population. Our results indicate a relative unawareness of people on the medicinal potential of the native species of their residence region. We suggest that strengthening connection between popular and scientific knowledge, along with spreading such knowledge, could contribute for an improved valuation about the Cerrado biome and consecutive preservation of it.KEYWORDS: Popular knowledge. Scientific knowledge. Valuation about the biome. INTRODUCTION Use of medicinal plants dates back to ancient times (DUTRA et al., 2016) and, over time, accumulated empirical evidence produced from popular knowledge has enabled incorporation of phytotherapy into traditional medicine with positive results (SANTOS et al., 2011). With the advent of ethnobotany and ethnopharmacology, many of these plants began to be safely used in the production ofherbal medicines for treatment of different diseases, such as infections, tissue inflammation, and pain (EKOR, 2014). It is currently known that several secondary metabolites produced by plants not only protect themselves from herbivory and pathogens, for instance (FÜRSTENBERG-HÄGG; ZAGROBELNY; BAK, 2013; SANCHÉZ-SANCHÉZ; MORQUECHO-CONTRERAS, 2017), but also have beneficial effects on human health (TAIZ; ZEIGER, 2010). However, in many countries, including those that hold a large biodiversity, the native medicinal flora is still barely studied by scientific methods (RIBEIRO et al., 2014). The Cerrado is the second largest biome after the Amazon in Brazil. Although the extensive area of 2,036,448 km2 occupied nowadays by this biome (BARBOSA, 2017), much of the Cerrado has already been transformed into pasture, grain crops or devastated for other uses (KLINK; MACHADO, 2005). Currently, only 8,21% of its total area is fully protected (BARBOSA, 2017). As a result, it is one of the most threatened biomes in the world. According to Mittermeier et al. (2005), the Brazilian Cerrado is one of the 25 global hotspots, areas with concentration of endemic species and exceptional ongoing destruction. Many communities resident in Cerrado regions are comprised of poor people with restricted access to public health services and often dependent Received: 14/05/18 Accepted: 20/11/19

O Cerrado é o segundo maior bioma do Brasil, apresenta uma diversidade ecológica imensurável e ainda pouco explorada. Apesar da exuberância de suas espécies endêmicas, é um dos 25 hotspots globais, pois apresenta alta riqueza natural em termos de biodiversidade e destruição expressiva de seu meio ambiente. Neste estudo, pesquisamos o conhecimento sobre o uso medicinal de plantas do Cerrado entre indivíduos que vivem em uma região predominantemente do Cerrado. Um questionário individual semiestruturado com perguntas de caráter socioeconômicas qualitativas e referentes ao uso medicinal de plantas foi aplicado aos usuários do Serviço de Saúde Pública da cidade de Assis, Estado de São Paulo, Brasil. Este estudo foi aprovado pelo Comitê de Ética do Hospital Regional de Assis (Protocolo 4812010). Dos 149 entrevistados, 90,6% relataram conhecimento e / ou uso de plantas para fins terapêuticos. Entre as 115 espécies de plantas mencionadas, 11,4% pertencem ao Cerrado. Também avaliamos a correlação entre os usos relatados e os respectivos dados publicados em estudos científicos sobre as propriedades medicinais das plantas.Verificou-se que, para as poucas plantas de Cerrado citadas, os entrevistados descreveram diversas aplicações medicinais não descritas cientificamente. Mais precisamente, 60,5% do total das aplicações medicinais não foram encontrados na literatura científica. Além disso, muitos dos efeitos terapêuticos descritos na literatura para as espécies do Cerrado não foram citados pela população entrevistada. Portanto, sugerimos que o fortalecimento da conexão entre conhecimento popular e científico, aliado à disseminação desse conhecimento, poderia contribuir para uma maior valorização do bioma Cerrado e consequentemente a preservação do mesmo.

Genetic diversity in section Rhizomatosae of the genus Arachis (Fabaceae) based on microsatellite markers

The genus Arachis (Fabaceae) native to South America, contains 80 species divided into nine sections, three of which contain species of special economic importance such as the cultivated peanut (Arachis hypogaea), belonging to the section Arachis, and some perennial forage species from sections Caulorrhizae and Rhizomatosae. We used microsatellite markers to assay genetic variability among 77 accessions of four species from section Rhizomatosae, the diploid Arachis burkartii (2n = 2x = 20) and the tetraploid Arachis glabrata, Arachis pseudovillosa and Arachis nitida (2n = 4x = 40). A total of 249 alleles were found in the fifteen loci analyzed and a high degree of intra and interspecific polymorphism was detected. The lowest intraspecific variation occurred in Arachis burkartii, while the smallest estimated interspecific value was between A. nitida and A. pseudovillosa and the largest was between A. burkartii and A. nitida. High observed heterozygosity was detected in A. glabrata. The diploid accessions grouped in one cluster and the tetraploid accessions in another. It was possible to distinguish all 77 accessions and the genetic distance between accessions could not be correlated with geographic origin.

Frequency and distribution of microsatellites from ESTs of citrus

Nearly 65,000 citrus EST (Expressed Sequence Tags) have been investigated using the CitEST project database. Microsatellites were investigated in the unigene sequences from Citrus spp. and Poncirus trifoliata. From these sequences, approximately 35 percent of the non-redundant ESTs contained SSRs. The frequencies of different SSR motifs were similar between Citrus spp and trifoliate orange. In general, mononucleotide repeats appeared to be the most abundant SSRs in the CitEST database, but we also identify di-, tri-, tetra-, penta- and hexanucleotide repeats. The AG/CT and AAG/CTT were the most common dinucleotide and trinucleotide motifs, with frequencies of 54.4 percent and 25.2 percent, respectively. Primer sequences flanking SSR motifs were successfully designed and synthesized. After in silico polymorphism analysis, a subset of sixty-eight primers was validated in different Citrus spp. and Poncirus trifoliata. PCR-amplification revealed polymorphism in citrus with all tested primer pairs and showed the potential of these markers for linkage mapping. Our study showed that the CitEST database can be exploited for the development of SSR markers that can amplify Citrus spp. and related genus for comparative mapping and other genetic analyses.