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Título: Flow cytometry reliability analysis and variations in sugarcane DNA content
Autor(es): Oliveira, A. C. L.
Pasqual, Moacir
Bruzi, Adriano Teodoro
Pio, Leila Aparecida Salles
Mendonça, P. M. S.
Soares, Joyce Dória Rodrigues
Assunto: Sugarcane – Varieties
Sugarcane – Genomes
Flow cytometry
Cana-de-açúcar – Variedades
Cana-de-açúcar – Genomas
Citometria de fluxo
Saccharum spp.
Publicador: Fundação de Pesquisas Científicas de Ribeirão Preto
Data de publicação: 29-Jun-2015
Referência: OLIVEIRA, A. C. L. et al. Flow cytometry reliability analysis and variations in sugarcane DNA content. Genetics and Molecular Research, Ribeirão Preto, v. 14, n. 2, p. 7172-7183, 2015.
Abstract: The aim of this study was to evaluate the reliability of flow cytometry analysis and the use of this technique to differentiate species and varieties of sugarcane (Saccharum spp) according to their relative DNA content. We analyzed 16 varieties and three species belonging to this genus. To determine a reliable protocol, we evaluated three extraction buffers (LB01, Marie, and Tris·MgCl2), the presence and absence of RNase, six doses of propidium iodide (10, 15, 20, 25, and 30 μg), four periods of exposure to propidium iodide (0, 5, 10, and 20 min), and seven external reference standards (peas, beans, corn, radish, rye, soybean, and tomato) with reference to the coefficient of variation and the DNA content. For statistical analyses, we used the programs Sisvar® and Xlstat®. We recommend using the Marie extraction buffer and at least 15 μg propidium iodide. The samples should not be analyzed immediately after the addition of propidium iodide. The use of RNase is optional, and tomato should be used as an external reference standard. The results show that sugarcane has a variable genome size (8.42 to 12.12 pg/2C) and the individuals analyzed could be separated into four groups according to their DNA content with relative equality in the genome sizes of the commercial varieties.
URI: http://www.geneticsmr.com/articles/4648
http://repositorio.ufla.br/jspui/handle/1/11508
Idioma: en_US
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