Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA

Abstract

Biomedicina -Faculdade Meta -FAMETA, Rio Branco, AC, Brasil.; 3. Professor, Doutor, Instituto Federal do Acre -IFAC, Cruzeiro do Sul, AC, Brasil.; 4. Analista da Empresa Brasileira de Pesquisa Agropecuária -Embrapa Acre, Rio Branco, AC, Brazil.; 5. Pesquisadora, Doutora, Empresa Brasileira de Pesquisa Agropecuária -Embrapa Rondônia, Porto Velho, RO, Brasil.; 6. Pesquisadora, Doutora, Empresa Brasileira de Pesquisa Agropecuária -Embrapa Acre, Rio Branco, AC, Brasil, tatiana.campos@embrapa.br. ABSTRACT:The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/µL (7 days in transport buffer) to 702.00 ng/µL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µL (30 days in silica gel) to 2,850.00 ng/µL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies.

Optimized method for dna extraction and PCR amplification in aroeira tree

“…Samples of young leaves of M. urundeuva were collected at the Center for Agricultural Sciences of the Federal University of Piauí (UFPI), transported in a Styrofoam box with ice, and storing them at a temperature of -20 ºC, until the start of DNA extractions. In general, the use of young leaves is recommended, as DNA with greater purity can be obtained due to the low concentrations of phenolic compounds in young leaves (Azêvedo et al 2019).…”

Comparação de métodos de isolamento do DNA em Handroanthus Serratifolius / Comparison of insulation methods of the DNA in Handroanthus Serratifolius

“…Coletou-se aproximadamente dez folhas jovens e raspas do caule de um exemplar de H. serratifolius localizado no Departamento de Biologia da Universidade Federal do Piauí, Teresina-PI. O uso de folhas jovens é recomendado pois esse tecido apresenta menos substâncias fenólicas que prejudicam a obtenção de um DNA puro (AZÊVEDO et al, 2019). Optou-se também pela utilização de raspas do caule pois indivíduos adultos de H. serratifolius podem atingir mais de 20 m de altura, o que pode dificultar a coleta das folhas na natureza.…”

Un estudio sobre el conocimiento científico que integra el sector de los fitoterápicos basados en la biodiversidad amazónica

“…En su mayoría, los artículos reportados en este grupo se enfocan en las características morfológicas, fisiológicas y ecológicas de las plantas amazónicas para un manejo sostenible de las florestas en pequeñas propiedades. Así, se encontraron artículos evaluando el coeficiente de repetibilidad en la producción de frutos de B. excelsa, (AZEVEDO et al, 2020) o diferentes métodos para la preservación de las hojas de E. precatoria para posteriores estudios moleculares (AZÊVEDO et al, 2019).…”