Trypsin inhibitor in Enterolobium contortisiliquum calli grown in the presence of plant growth regulators

Abstract

The objective of this work was to investigate the effect of plant growth regulators (PGR) on the induction of calli from cotyledons of Enterolobium contortisiliquum grown in vitro, as well as to evaluate the inhibitory activity of trypsin in these calli. Cotyledons of E. contortisiliquum were cultivated in culture medium supplemented with dichlorophenoxyacetic acid (2,4-D), picloram (PIC), kinetin (KIN), and 6-benzylaminopurine (BAP). Callus mass, inhibitory activity of trypsin, morphogenetic and cytochemical observations, and biological activity were measured after 60 days. The Pareto diagram showed that only 0.5 mg L^-1 PIC influenced the increase of fresh mass, whereas 0.5 mg L^-1 BAP influenced dry mass. The principal component analysis plot showed the formation of two clear clusters. The inhibitory activity of trypsin is achieved at a higher intensity in the treatment with 0.5 mg L^-1 2,4-D + 2.0 mg L^-1 PIC + 0.5 mg L^-1 KIN + 2.0 mg L^-1 BAP. The combination of PGR at different rates causes an intense callus formation. Low concentrations of PGR result in a higher fresh mass, dry mass, and biological activity. The use of PGR is efficient in obtaining E. contortisiliquum calli with a satisfactory inhibitory activity of trypsin.