In vitro conservation and low cost micropropagation of Cochlospermum regium (Mart. Ex. Scharank)

Cochlospermum regium (Mart. Ex. Scharank), a Cochlospermaceae is a Cerrado medicinal species showing antimicrobial activity on the female reproductive system. The species risks being genetically eroded since it is highly explored, the biome devastated and the plant roots are employed for phytotherapeutic preparations. This study aims to develop a micropropagation protocol for large scale production of plantlets and establish the species conservation in a germplasm bank, in vitro. Nodal segments were inoculated into Murashige and Skoog (MS) medium supplemented with different concentrations of sucrose, BAP, kinetin, zeatin, Phytagel (R) and activated charcoal. The bud position effect on the multiplication index was evaluated and the auxin IBA utilized in rooting experiments. The best multiplication response was with the apical bud in culture medium in absence of cytokinins. MS inoculated explants with different IBA concentrations did not root in vitro, but ex vitro rooting was satisfactory. Survival of explants, 47.62%, was for 4 months in germplasm bank conditions without culture medium transference.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Histochemical investigation of Cochlospermum regium (Schrank) Pilg. leaves and chemical composition of its essential oil

Essential oil from Cochlospermum regium (Schrank) Pilg. leaves (CR-EO) has been extracted by hydrodistillation; we analysed the CR-EO by gas chromatography coupled with mass spectrometry. We also conducted histochemical analysis on cross-sections of the central vein of young and adult leaves. A total of 32 compounds were qualitatively and quantitatively analysed, which represented 94.87% of the total CR-EO oil content. The CR-EO basically consisted of sesquiterpenes (96.87%); its main component was beta-copaen-4-alpha-ol (18.73%), followed by viridiflorol (12.67%). The histochemical analyses identified the main classes of compounds present in both young and adult leaves.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Genetic diversity of Stryphnodendron adstringens (Mart.) Coville determined by AFLP molecular markers

Bark extracts of Stryphnodendron adstringens (Mart) Coville a Leguminosae species, well known in Brazil as barbatimao, are popularly used as healing agent. The objective of this work was to determine the genetic diversity of S. adstringens populations and to correlate genetic distances to the production of tannins. S. adstringens accessions from populations found in Cerrado regions in the states of Goias, Minas Gerais and São Paulo were analyzed using the AFLP (Amplified Fragment Length Polymorphism) technique. A total of 236 polymorphic bands were scored and higher proportion of genetic diversity was found inter populations (70.9%), rather than intra populations (29.1%). F-ST value was found to be significantly greater than zero (0.2906), demonstrating the complex genetic structure of S. adstringens populations. Accessions collected in Cristalina, GO, showed higher percentage of polymorphic loci (87.3%) and the highest genetic diversity. The lowest genetic variability was detected among accessions from the population growing in Caldas Novas, GO. The genetic distance among populations was estimated using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA), which grouped populations into 3 clusters. Moreover, chemotypes with tannin concentration above 40% showed higher genetic similarity. AFLP analysis proved to be an efficient gene mapping technique to determine the genetic diversity among remaining populations of S. adstringens. Obtained results may be employed to implement further strategies for the conservation of this medicinal plant. (C) 2011 Elsevier Ltd. All rights reserved

Chloroplast genome characterization of Uncaria guianensis and Uncaria tomentosa and evolutive dynamics of the Cinchonoideae subfamily

Abstract Uncaria species are used in traditional medicine and are considered of high therapeutic value and economic importance. This work describes the assembly and annotation of the chloroplast genomes of U. guianensis and U. tomentosa, as well as a comparative analysis. The genomes were sequenced on MiSeq Illumina, assembled with NovoPlasty, and annotated using CHLOROBOX GeSeq. Addictionaly, comparative analysis were performed with six species from NCBI databases and primers were designed in Primer3 for hypervariable regions based on the consensus sequence of 16 species of the Rubiaceae family and validated on an in-silico PCR in OpenPrimeR. The genome size of U. guianensis and U. tomentosa was 155,505 bp and 156,390 bp, respectively. Both Species have 131 genes and GC content of 37.50%. The regions rpl32-ccsA, ycf1, and ndhF-ccsA showed the three highest values of nucleotide diversity within the species of the Rubiaceae family and within the Uncaria genus, these regions were trnH-psbA, psbM-trnY, and rps16-psbK. Our results indicates that the primer of the region ndhA had an amplification success for all species tested and can be promising for usage in the Rubiaceae family. The phylogenetic analysis recovered a congruent topology to APG IV. The gene content and the chloroplast genome structure of the analyzed species are conserved and most of the genes are under negative selection. We provide the cpDNA of Neotropical Uncaria species, an important genomic resource for evolutionary studies of the group

Genetic structure and chemical diversity in natural populations of Uncaria guianensis (Aubl.) J.F.Gmel. (Rubiaceae).

Uncaria guianensis is native to the Amazon and is used traditionally as an anti-inflammatory. Natural populations of the species have declined markedly in recent times because of strong anthropic pressure brought about by deforestation and indiscriminate collection. The aim of the present study was to assess the genetic and chemical diversity among eight natural populations of U. guianensis located in the Brazilian states of Acre, Amapá and Amazonas. A set of four primer combinations was employed in sequence-related amplified polymorphism (SRAP) amplifications of leaf DNA, and the fragments were analyzed in an LI-COR model 4300 DNA Analyzer. Genetic variability within the populations (81%) was substantially greater than that detected between them (19%). The highest percentage of polymorphic loci (90.21%) and the largest genetic variability were observed in the population located in Mazagão, Amapá. Genetic differentiation between populations was high (Fst = 0.188) and the studied populations formed three distinct genetic groups (K = 3). The population located in Assis Brasil, Acre, presented the highest average content of the mitraphylline (0.60 mg/g dry weight,). However, mitraphylline and isomitraphylline not detected in most individuals in the studied populations, and it is questionable whether they should be considered as chemical markers of the species. The genetic data confirm the urgent need for conservation programs for U. guianensis, and for further studies aimed at ascertaining the genetic basis and heritability of alkaloid accumulation

Sampling sites of <i>Uncaria tomentosa</i> populations located in the Amazon region of Brazil.

<p>The sites of collection of the eight populations collected found in the states of Acre (AC), Amapá (AP) and Pará (PA) are showed on the map (1–8), 1- Mâncio Lima, AC; 2- Cruzeiro do Sul, AC; 3- Tarauacá, AC 4-Feijó, AC; 5- Mazagão, AP; 6- Santana, AP; 7- Macapá, AP and 8-Afuá, PA. Populations with the largest (1) and smallest (3) genetic variability are evinced by white circles. (Map source: <a href="https://eros.usgs.gov/" target="_blank">https://eros.usgs.gov/</a>).</p

Dendrogram, constructed using the unweighted pair group method with arithmetic mean (UPGMA), showing eight populations of <i>Uncaria tomentosa</i> separated into two clusters.

<p>Group I contained UTTA (Tarauacá, Acre), UTFJ (Feijó, Acre) and UTMC (Macapá, Amapá), while group II comprised UTMZ (Mazagão, Amapá), UTML (Mâncio Lima, Acre), UTCS (Cruzeiro do Sul, Acre), UTSA (Santana, Amapá) and UTAF (Afuá, Pará).</p

Genetic parameters of the populations of <i>Uncaria tomentosa</i> collected in the Amazon region of Brazil.

<p>Genetic parameters of the populations of <i>Uncaria tomentosa</i> collected in the Amazon region of Brazil.</p

High performance liquid chromatographic (HPLC) analyses of leaf extracts of three specimens of <i>Uncaria tomentosa</i> populations from the Amazon region of Brazil.

<p>The chromatograms were obtained from specimens collected in: (<b>A</b>) Cruzeiro do Sul, AC, in which peaks labeled <b>a</b> and <b>b</b> correspond to mitraphylline and isomitraphylline, respectively; (<b>B</b>) Afuá, PA, in which the peak labeled <b>b</b> corresponds to isomitraphylline;; (<b>C</b>) Mâncio Lima, AC, in which peaks <b>a</b> and <b>b</b> are absent. Standards (D) mitraphylline and (E) isomitraphylline.</p