Comparison of protocols for genomic DNA extraction from ‘velame pimenta’ (Croton linearifolius), a native species to the Caatinga, Brazil

The Caatinga biome occupies some 12% of the Brazilian territory, which is present in at least nine states. The species that constitute its biodiversity have the potential to be used as natural resources, among them are approximately 700 species of the genus Croton. As an example of this potential, the Croton linearifolius specie is used by local communities as a natural insecticide. Associated with the economic potential of the Caatinga species, one must stress the risk of extinction or genetic erosion due to the growing deforestation of natural areas of this biome. These factors make it relevant in genetic studies in order to guide conservation strategies. Considering the lack of molecular studies involving C. linearifolius, we compared the efficiency of six protocols for genomic DNA extraction previously described in literature. The DNA extraction buffers [based on the use of Cetyl trimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), mannitol and sorbitol] were different in their efficiency to obtaining the genomic DNA of C. linearifloius. In general, protocols using CTAB buffer were more efficient. The use of liquid nitrogen in the maceration process was also evaluated and its use was considered a no necessary factor in obtaining DNA in adequate quantity and quality for PCR platform procedures.Keywords: DNA Isolation, molecular markers, native species of CaatingaAfrican Journal of Biotechnology Vol. 12(30), pp. 4761-476

Comparison of coefficients and distance measurements in passion fruit plants based on molecular markers and physicochemical descriptors

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)We investigated seven distance measures and 14 similarity coefficients in a set of observations of variables of the 'yellow' passion fruit plant (Passiflora edulis Sims), submitted to multivariate analyses (distance, projection and grouping). Fourteen genotypes were characterized, based on DNA amplification with 16 random amplified polymorphic DNA primers and the assessment of nine fruit physical-chemical descriptors. The distance measurements and the similarity coefficients were compared by the Spearman correlation test, projection in two-dimensional space and grouping efficiency, using five grouping methods; the genotype ranking varied with the different techniques. Coler-Rodger distance measures, Euclidean distance square measures and Yule similarity coefficients proved to be inadequate for projection in two-dimensional space or for grouping matrices. Regardless of the origin of the distance matrix, the unweighted pair group method with arithmetic mean grouping method proved to be the most adequate. The various distance measurements, similarity coefficients and grouping methods gave different values of distortion, cophenetic correlation and stress; they influence the characterization of genetic variability and this should be taken into account for studies of yellow passion fruit plants.83870879Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Programa de Pos-Graduacao em Genetica e Biologia Molecular, UESCConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Genetic variability in wild genotypes of Passiflora cincinnata based on RAPD markers

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The genetic diversity and characteristics of commercial interest of Passiflora species make it useful to characterize wild germplasm, because of their potential use for fruit, ornamental and medicinal purposes. We evaluated genetic diversity, using RAPD markers, of 32 genotypes of Passiflora cincinnata collected from the wild in the region of Vitoria da Conquista, Bahia, Brazil. Thirteen primers generated 95 polymorphic markers and only one monomorphic marker. The mean genetic distance between the genotypes estimated by the complement of the Dice index was 0.51 (ranging from 0.20-0.85), and genotype grouping based on the UPGMA algorithm showed wide variability among the genotypes. This type of information contributes to identification and conservation of the biodiversity of this species and for the identification of pairs of divergent individuals for maximum exploitation of existing variability.9424212428Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)UESCConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

New polymorphic microsatellite loci for Theobroma cacao: isolation and characterization of microsatellites from enriched genomic libraries

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Seventeen polymorphic microsatellite markers were isolated from enriched genomic libraries for Theobroma cacao, providing additional tools for studying the genetic diversity and map saturation of this species. These markers were characterized in 32 accessions of the T. cacao germplasm collection from the Centro de Pesquisas do Cacau. The number of alleles at each locus varied from 2 to 8, with an average of 4.41 alleles per locus. The polymorphism information content varied from 0.060 to 0.695, with an average of 0.333. The markers characterized in this study will be employed in map saturation studies and diversity assessments of cacao genotypes.564789792Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CAPES [PROCAD-NF2008

Genetic variation in a wild population of the 'sleep' passion fruit (Passiflora setacea) based on molecular markers

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Little is known about the molecular genetic diversity of most Passiflora species. We used RAPD markers to evaluate the genetic diversity of 24 genotypes of the 'sleep' passion fruit (Passiflora setacea). Twelve primers generated 95 markers, 88% of which were polymorphic. The genetic distance estimated by the complement of the Dice index ranged from 0.29 (among accessions Ps-G1 and Ps-G13) to 0.69 (among accessions Ps-G21 and Ps-G23). Genotype grouping based on the UPGMA algorithm showed considerable variability among genotypes. We conclude that P. setacea has a broad genetic base that could be exploited in breeding programs.111731738Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)UESCConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Genetic diversity in wild species of passion fruit (Passiflora trintae) based on molecular markers

In spite of the importance of and the considerable variability observed in Passiflora (Passifloraceae), little is known about the genetic diversity of most of the species of this genus. We evaluated the genetic diversity by RAPD markers in 18 genotypes of Passiflora trintae. The 15 primers generated 112 markers, 84% of which were polymorphic. The genetic distance estimated by the complement of the Dice index (average dissimilarity = 0.30) and genotype grouping based on the UPGMA algorithm showed low variability among genotypes. More attention should be given to the study and conservation of the biodiversity of this economically important genus.942123213

DEVELOPMENT AND CHARACTERIZATION OF MICROSATELLITE MARKERS FOR THE WILD SOUTH AMERICAN PASSIFLORA CINCINNATA (PASSIFLORACEAE)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Premise of the study: We developed the first set of microsatellite markers for the passion fruit, Passiflora cincinnata, to provide tools for further study of its genetic diversity and to support current conservation and genetic studies. Methods and Results: We used a microsatellite-enriched library approach to isolate and characterize 25 new molecular markers. Seven primer pairs successfully amplified polymorphic microsatellite regions, with an average of five alleles per locus. The mean values of expected and observed levels of heterozygosity were 0.516 and 0.525, respectively. Conclusions: The microsatellite markers identified in this study may be valuable tools for population genetic studies, and this set of markers also may be useful in the design of a genetic conservation strategy, mating system, and hybridization studies.994E170E172Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)CAPES [PROCAD-NF2008