Partial characterization of genes from the embryonic axis of Melanoxylon brauna Schott. (Leguminosae-Caesalpinioideae) seeds

The objective of this study was to partially characterize some genes involved in the desiccation tolerance of the embryonic axis of Melanoxylon brauna seeds subjected, or not, to oven fast-drying. Seeds were initially dried rapidly in an oven at 40 ºC, 50 ºC, 60 ºC, 70 ºC, and 80 °C, for 24, 48 and...

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Published in:Revista brasileira de sementes 2012, Vol.34 (1), p.29-38
Main Authors: Silva, Mirian de Sousa, Brommonschenkel, Sérgio Hermínio, Faria, José Márcio Rocha, Borges, Eduardo Euclydes de Lima e
Format: Article
Language:eng ; por
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PLANT SCIENCES
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Summary:The objective of this study was to partially characterize some genes involved in the desiccation tolerance of the embryonic axis of Melanoxylon brauna seeds subjected, or not, to oven fast-drying. Seeds were initially dried rapidly in an oven at 40 ºC, 50 ºC, 60 ºC, 70 ºC, and 80 °C, for 24, 48 and 72 h and then subjected to germination tests and moisture content determination. Degenerate primers were designed for 19 genes. The CDNA was used as a template for PCR amplifications using the degenerate primers, and the PCR products obtained were purified, cloned and sequenced. The seeds showed a gradual reduction in percent germination with increasing temperature and drying time. Nucleotide sequences of the cloned fragments related to genes CAT1, SPS1, Abi5, Transk and PM25 were obtained. The similarity analysis with the sequences deposited in databases revealed similarities with genes CAT1, SPS1, Transk and PM25 from other plant species. The nucleotide sequences obtained from the respective genes will be used for designing specific primers for gene expression analyses during seed germination in order to understand the causes for loss of physiological quality of Melanoxylon brauna seeds. O presente trabalho objetivou caracterizar parcialmente os genes envolvidos com a tolerância à dessecação do eixo embrionário de sementes de Melanoxylon brauna submetidas ou não à secagem rápida. As sementes foram submetidas à secagem rápida em estufa nas temperaturas de 40 ºC, 50 ºC, 60 ºC, 70 ºC e 80 ºC, nos tempos de 24, 48 e 72 h e, em seguida, submetidas ao teste de germinação e determinação do grau de umidade. Foram utilizados primers degenerados para 19 genes. Os produtos de PCR obtidos foram purificados, clonados no vetor pGEM-T Easy e sequenciados. As sementes apresentaram redução gradativa na porcentagem de germinação com o aumento da temperatura e tempo de secagem. Foram obtidas as sequências dos fragmentos clonados referentes aos genes CAT1, SPS1, ABI5, Transk e PM25. A comparação das sequências obtidas com sequências depositadas em bancos de dados revelou similaridade com os genes CAT1, SPS1, Transk e PM25 de outras espécies. As sequências de nucleotídeos obtidas dos respectivos genes servirão para a confecção de oligonucleotídeos específicos para posteriores estudos de expressão gênica durante a germinação de sementes de M. brauna submetidas à secagem, visando entender as causas da perda de qualidade fisiológica dessas sementes.
ISSN:0101-3122
0101-3122
DOI:10.1590/S0101-31222012000100004