Validation and finding of distant orthologous genes of the DAF-12 gene in Meloidogyne Incognita using bioinformatics tools

BETANZOS SAN JUAN, RAFAEL; SCHUSTER, CLAUDIO DAVID; MODENUTTI, CARLOS PABLO

Congreso; XII CAB2C (Congreso Argentino de Bioinformatica y Biologia Computacional); 2022

Asociacion Argentina de Bioinformatica y Biologia Computacional

Root-knot nematode (RKN) Meloidogyne spp. is one of the most damaging parasites due to its wide range of hosts. Here, we report a DAF-12 homologue from Meloidogyne incognita (DAF-12Minc) that had not been described before and that is a promising target to control infestations. Using a combination of Hidden Markov Models (HMM) based sequence search and phylogenetic analysis we identify three orthologous of C. elegans receptor DAF-12 in M. incognita. These candidates displayed high identity percentages between themselves, indicating that it could come from a triplication event. Despite the global sequence identity between previous DAF-12 orthologous and DAF-12Minc were acceptable, the key amino acids in the hormone receptor binding site were missing. In that sense, we evaluated if our DAF-12Minc candidates were able to bind Dafachronic acids (DA’s) by modeling the structure of DAF-12Minc ligand-binding domain and performed Molecular Dynamics to refine the models, pocket detection to identify the ligand binding site of the protein and molecular docking to evaluate previously reported ligands. These analyses suggested that the features of the active site are conserved. Moreover, the luciferase reporter gene led us to elucidate the antagonistic effects of the DA’s; and further in vitro and in vivo analysis suggest that DA’s had an effect in both life cycle and mortality of M. incognita, produces an overall improvement in plant health condition due to the effect of the compound on M. incognita life cycle, and that DA’s could have an effect not only in the infection by M. incognita, but also a long-term effect in the inhibition of offspring hatching.