Exsudatos de sementes de soja [Glycine max (L.) Merrill]: caracterização proteômica e efeitos contra o nematoide das galhas Meloidogyne incognita

Abstract

Exudation is a defense mechanism presented by seeds, which release molecules to the soil during the germination process. These exudates, mainly composed by proteins and secondary metabolites, act interfering with the development of microorganisms. They represent a source of defense molecules with biotechnological potential to control soil-borne pathogens. In this context, soybean [Glycine max (L.) Merrill] stands as an ideal system in the study of plant-pathogen interaction due to its great economic value and the high content of seed proteins. The experimental activities presented in this thesis aimed to characterize the composition of seed exudates from two different soybean cultivars, which show different levels of performance in response to the root-knot nematode Meloidogyne incognita. Furthermore, the in vitro and in vivo effects of the seed exudates on this pathogen were evaluated. For this purpose, the exudates were obtained from soybean seeds (BRS-Pala:susceptible; BRS-Macota:resistant), soaked in 0.1 M sodium acetate, pH 5.0, during 18 hours. The gel-free proteomic analysis allowed identification of several defense-related proteins such as soybean agglutinin, Kunitz and Bowman-Birk trypsin inhibitors, and pathogenesis-related proteins. The qualitative analysis of secondary metabolites in the exudates also revealed the presence of phenols, triterpenoids, saponins, flavones, flavonols, and xanthones in both cultivars. The above exudates were dialyzed in 3.5 kDa cut-off membranes and assayed for bioactive proteins. The hemagglutinating (609.52 HU/mgP), trypsin (4.81 mg trypsin inhibited/mgP) and papain (30.48 IU/mgP) inhibitor, proteolytic (1.52 UAPr/mgP.min), and β-1, 3-glucanase (58.26 ηkatGLU/mgP) activities were higher for BRS-Pala. Otherwise, the peroxidase activity (0.218 UAP/mgP.min) predominated in BRS-Macota. The evaluation of anti-nematode activity against eggs and second stage juveniles (J2) revealed that both exudates were able to inhibit the hatching of M. incognita eggs as well as the J2 mobility. This nematostatic property is reflected in the infectivity of the parasite, reduced by the exuded protein at 0.13 mg/mL concentration. The results reinforce the idea that the exudation is related to the constitutive defense of the seeds, suggesting that the proteins present in the exudates can contribute to the protection against nematodes and possibly other plant enemies during the germination process.