Improvements in cytological preparations for fluorescent in situ hybridization in Passiflora

dc.creatorSouza, MM
dc.creatorUrdampilleta, JD
dc.creatorForni-Martins, ER
dc.date2010
dc.date2014-11-14T16:07:30Z
dc.date2015-11-26T17:15:48Z
dc.date2014-11-14T16:07:30Z
dc.date2015-11-26T17:15:48Z
dc.date.accessioned2018-03-29T00:04:00Z
dc.date.available2018-03-29T00:04:00Z
dc.identifierGenetics And Molecular Research. Funpec-editora, v. 9, n. 4, n. 2148, n. 2155, 2010.
dc.identifier1676-5680
dc.identifierWOS:000284324400016
dc.identifier10.4238/vol9-4gmr951
dc.identifierhttp://www.repositorio.unicamp.br/jspui/handle/REPOSIP/69274
dc.identifierhttp://www.repositorio.unicamp.br/handle/REPOSIP/69274
dc.identifierhttp://repositorio.unicamp.br/jspui/handle/REPOSIP/69274
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/1282131
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.descriptionCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.descriptionCytological preparations for the fluorescent in situ hybridization (FISH) technique require cytoplasm-free metaphases, with well-spread chromosomes, for the localization of DNA sequences and chromosome mapping. We tested various procedures for FISH analysis of Passiflora cacaoensis, P. gardneri and hybrid F(1) progeny of P. gardneri x P. gibertii. Two treatments with four enzymes and three incubation times were compared. The material was treated with 1.0 M HCl before enzymatic digestion. The following criteria were used to determine the quality of the metaphases: a) lack or presence of cytoplasm; b) well-spread chromosomes or with overlap; c) complete or incomplete chromosome number (2n). The enzyme Pectinex (R) SP ULTRA gave the best performance, with the shortest incubation time. The best results were observed after 30 min of incubation; more than 70% of the metaphases did not have large amounts of cytoplasm or overlapping chromosomes, and about 75% maintained the chromosome number. FISH was carried out using a 45S rDNA probe (pTa71) labeled with biotin and detected with fluorescein isothiocyanate. Sites with strong staining and without nonspecific signals were observed. Our methodological adaptations allowed the preparation of metaphase slides of high quality for the FISH technique, with less time required for the preparation of samples.
dc.description9
dc.description4
dc.description2148
dc.description2155
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionFundacao de Amparo a Pesquisa do Estado da Bahia (FAPESB)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.descriptionUniversidade Estadual de Santa Cruz (UESC)
dc.descriptionCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.descriptionPrograma de Pos-Graduacao em Producao Vegetal (PPGPV/UESC)
dc.descriptionFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.descriptionConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.descriptionCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.languageen
dc.publisherFunpec-editora
dc.publisherRibeirao Preto
dc.publisherBrasil
dc.relationGenetics And Molecular Research
dc.relationGenet. Mol. Res.
dc.rightsfechado
dc.sourceWeb of Science
dc.subjectPassion flower
dc.subjectCytogenetic
dc.subjectPectinex
dc.subjectFISH
dc.subjectChromosome-numbers
dc.subject45s Rdna
dc.subjectHybrids
dc.subjectGenome
dc.subjectLinkage
dc.subjectWheat
dc.subjectMap
dc.titleImprovements in cytological preparations for fluorescent in situ hybridization in Passiflora
dc.typeArtículos de revistas


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