In silico characterization and expression analysis of the multigene family encoding the Bowman–Birk protease inhibitor in soybean
The Bowman–Birk (BBI) protease inhibitors can be used as source of sulfur amino acids, can regulate endogenous protease activity during seed germination and during the defense response of plants to pathogens. In soybean this family has not been fully described. The goal of this work was to characterize in silico and analyze the expression of the members of this family in soybean. We identified 11 potential BBI genes in the soybean genome. In each one of them at least a characteristic BBI conserved domain was detected in addition to a potential signal peptide. The sequences have been positioned in the soybean physical map and the promoter regions were analyzed with respect to known regulatory elements. Elements related to seed-specific expression and also to response to biotic and abiotic stresses have been identified. Based on the in silico analysis and also on quantitative RT-PCR data it was concluded that BBI-A, BBI-CII and BBI-DII are expressed specifically in the seed. The expression profiles of these three genes are similar along seed development. Their expressions reach a maximum in the intermediate stages and decrease as the seed matures. The BBI-DII transcripts are the most abundant ones followed by those of BBI-A and BBI-CII.
KeywordsBBI Glycine max qRT-PCR
This work was funded by CNPq, FAPEMIG and FINEP (Brazilian Government). B.A. Barros was the recipient of a PhD fellowship from CNPq.
- 6.Fernadez JH, Mello MO, Galgaro L, Tanaka AS, Silva-Filho MC, Neshich G (2007) Proteinase inhibition using small Bowman–Birk-type structures. Genet Mol Res 6:846–858Google Scholar
- 14.MacDonald MB, Elliot IJ, Sweeney PM (1999) DNA extraction from dry seeds for RAPD analyses in varietal identification studies. Seed Sci Technol 22:171–176Google Scholar
- 15.Sambrook J, Frietsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor, New YorkGoogle Scholar
- 21.Horisberger M, Tacchini-Vonlanthen M (1983) Ultrastructural localization of Bowman–Birk inhibitor on thin sections of Glycine max (soybean) cv Maple Arrow by the gold method. Histochem Cell Biol 3:313–321Google Scholar
- 28.Rakwal R, Agrawal GK, Jwa N-S (2001) Characterization of a rice (Oryza sativa L.) Bowman–Birk proteinase inhibitor: tightly regulated induction in response to cut, jasmonic acid, ethylene and protein phosphatase 2A inhibitors. Gene 263:189–198. doi: 10.1016/S0378-1119(00)00573-4 PubMedCrossRefGoogle Scholar
- 33.Goldberg RB, Crouch ML, Walling L (1983) Regulation of soybean seed protein gene expression. In: Nagley P, Linnane AW, Peacok WJ, Pateman JA (eds) Manipulation and expression of genes in eukaryotes. Academic Press, New York, pp 193–201Google Scholar