β-galactosidases from cowpea stems: properties and gene expression under conditions of salt stress

Fabrício Bonfim Sudério, Enéas Gomes Filho, José Hélio Costa, Joaquim Enéas Filho

Resumo


The aim of this work was to evaluate the properties and gene expression of isoforms of β-galactosidase, isolated from the stems of seedlings of the cowpea cv. Pitiuba, at different stages of development and sown in the absence and presence of NaCl. Seeds were sown onto sheets of filter paper soaked in distilled water (control) and a 100 mM NaCl solution. The plants were collected at the III (ED III) and VIII (ED VIII) stages of development. Isoforms of the enzyme β-gal I, II and III presented molecular weights of 89, 146 and 124 kDa respectively, and the same pH (4.0) and optimum test temperature (55 °C) under all the conditions studied. The three isoforms were stable up to 50 °C. The thermal inactivation curves of the enzymes as a function of time were biphasic at a temperature of 60 °C. Analysis of the gene expression of β-galactosidase was carried out using degenerate primers and total RNA from the stems of seedlings at ED III and ED VIII, sown both as control and under saline treatments. The results showed higher transcript levels at ED VIII, with reduced levels of β-galactosidase transcripts under saline treatment, and that β-gal I, II and III do not undergo changes during the early stages of development. There are a few differences in the properties of the three isoforms. Salinity affected the gene expression of enzymes. The enzymes were expressed differently for ED III and ED VIII.


Palavras-chave


Vigna unguiculata; Enzymes; Purification

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Referências


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