ABSTRACT. Molecular markers are important for characterizing the genetic diversity of plants and can provide the basis for strategies to protect and conserve endangered populations. However, numerous molecular techniques are used, requiring an evaluation of fast and efficient methods to extract DNA. Since molecular studies of Caesalpinia ferrea are rare, it is important to develop and/or adapt a DNA extraction protocol that produces quality DNA samples to enable the design of strategies for the conservation of this threatened species. This study aimed to compare five methods for DNA extraction and to determine the most efficient protocol for C. ferrea. Sufficient genomic DNA was obtained from the leaves of C. ferrea using all the tested protocols to perform techniques DNA extraction from Caesalpinia ferrea involving molecular markers. Two protocols based on the detergent cetyl trimethyl ammonium bromide, as well as a commercial kit, yielded high concentrations of pure DNA. However, when polymerase chain reaction amplifications were performed, DNA was only successfully amplified from extractions performed with the commercial kit, which produced sufficient genomic DNA of good quality from the leaves of C. ferrea to perform techniques involving molecular markers.
RESUMO -O bacurizeiro é uma das espécies fruteiras nativas de maior importância socioeconômica das regiões Norte e Meio-Norte do Brasil. No entanto, ainda há carência de conhecimentos científicos sobre a espécie, especialmente sobre sua biologia floral, mecanismos reprodutivos e viabilidade do pólen. O conhecimento sobre a viabilidade do pólen é fundamental para o melhoramento genético, especialmente quando se tem em mente a realização de hibridizações controladas. O objetivo deste trabalho foi avaliar o efeito da sacarose e de acessos na viabilidade do pólen de bacurizeiro por meio da germinação in vitro. Avaliaram-se cinco acessos (BGB 6, BGB 11, BGB 48, BGB 32 e BGB 16) de bacurizeiro combinados com um tratamento-controle, sem sacarose, e quatro concentrações de sacarose (5; 7,5; 10 e 20%), em delineamento inteiramente ao acaso, em arranjo fatorial 5 x 5, com oito repetições. Na maioria dos acessos, a germinação in vitro do pólen atingiu um máximo (71,1 e 63,0%) entre as concentrações de sacarose de 9,4 e 9,7% e, a partir daí, caiu de forma drástica até atingir um percentual inferior a 10% de germinação na concentração de 20% de sacarose. Foram encontrados bons percentuais de germinação (entre 63,0 e 77,1%) para todos os acessos, com exceção do acesso BGB 16, que não alcançou 50%. Palavras-chave: Clusiaceae, viabilidade do pólen, melhoramento genético, meio de cultura, pólen fresco. EFFECT OF SUCROSE CONCENTRATION ON POLLEN IN VITRO GERMINATION OF FIVE BACURY (Platonia insignis MART.) ACCESSESABSTRACT -The bacury tree is a native fruit species of great socio-economic importance of North and Mid-North regions of Brazil. However, there is insufficient scientific knowledge on the species, especially on its floral biology, reproductive mechanisms and pollen viability. Knowledge about pollen viability is essential for the genetic breeding, especially for controlled hybridizations. The aim of this study was to evaluate the effect of sucrose and of access on the viability of bacury pollen by in vitro germination. Five bacury accessions (BGB 6, BGB 11, BGB 48, BGB 32 and BGB 16) and five sucrose concentrations (0, 5, 7.5, 10, and 20%) were evaluated in a completely randomized design in a 5 x 5 factorial arrangement, with eight replications. In the majority of the accessions, pollen germination in vitro reached a maximum (71.1 to 63.0%) sucrose concentrations between 9.4 and 9.7% and from then fell dramatically until a percentage lower than 10% germination at a concentration of 20% sucrose. Showed good germination percentages (between 63.0 and 77.1%) for all accessions, except access BGB 16 did not reach 50%.
ABSTRACT. Cratylia argentea (Desv.) Kuntze (Fabaceae) is a droughttolerant, perennial legume found primarily in Brazil, Bolivia, and Peru. The shrub is well adapted to acid soils and exhibits high productivity and nutritional value, characteristics that would favor its use as a dry season animal forage supplement in semiarid regions. In plant improvement programs, the production of elite hybrids with superior traits is generally achieved by crossing parents that exhibit the highest level of genetic divergence. Therefore, the aim of the present study was to assess genetic diversity among 13 accessions of C. argentea from the same population maintained in the active germplasm bank of Embrapa Meio-Norte using inter-simple sequence repeat (ISSR) markers. Genetic similarities between C. argentea accessions were estimated from Jaccard coefficients, and a dendrogram was constructed using the unweighted pair group method with arithmetic average (UPGMA). The set of 15 primers selected for ISSR analysis generated a total of 313 loci of which 79.23% were polymorphic. The mean number of bands per primer was 20.87, and the amplicons 15243 Genetic diversity among accessions of Cratylia argentea ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (4): 15242-15248 (2015) ranged from 280 to 3000 bp in size. Primers UBC834 and UBC827 generated the largest number of polymorphic loci and exhibited 90.91 and 100% polymorphism, respectively. The coefficients of genetic similarity among accessions varied between 0.49 and 0.73. UPGMA cluster analysis allowed the identification of four genotypic groups and demonstrated the existence of considerable variability within the collection. Potential progenitors were selected that would offer good possibilities of obtaining unusual and favorable combinations of genes in a plant breeding program.
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