Flagellin and mannitol modulate callose biosynthesis and deposition in soybean seedlings.

Callose is a polymer deposited on the cell wall and is necessary for plant growth and development. Callose is synthesized by genes from the glucan synthase-like family (GSL) and dynamically responds to various types of stress. Callose can inhibit pathogenic infection, in the case of biotic stresses, and maintain cell turgor and stiffen the plant cell wall in abiotic stresses. Here, we report the identification of 23 GSL genes (GmGSL) in the soybean genome. We performed phylogenetic analyses, gene structure prediction, duplication patterns, and expression profiles on several RNA-Seq libraries. Our analyses show that WGD/Segmental duplication contributed to expanding this gene family in soybean. Next, we analyzed the callose responses in soybean under abiotic and biotic stresses. The data show that callose is induced by both osmotic stress and flagellin 22 (flg22) and is related to the activity of ß-1,3-glucanases. By using RT-qPCR, we evaluated the expression of GSL genes during the treatment of soybean roots with mannitol and flg22. The GmGSL23 gene was upregulated in seedlings treated with osmotic stress or flg22, showing the essential role of this gene in the soybean defense response to pathogenic organisms and osmotic stress. Our results provide an important understanding of the role of callose deposition and regulation of GSL genes in response to osmotic stress and flg22 infection in soybean seedlings.

Methyl-CpG binding proteins (MBD) family evolution and conservation in plants.

DNA methylation is an epigenetic mechanism that acts on cytosine residues. The methyl-CpG-binding domain proteins (MBD) are involved in the recognition of methyl-cytosines by activating a signaling cascade that induces the formation of heterochromatin or euchromatin, thereby regulating gene expression. In this study, we analyzed the evolution and conservation of MBD proteins in plants. First, we performed a genome-wide identification and analysis of the MBD family in common bean and soybean, since they have experienced one and two whole-genome duplication events, respectively. We found one pair of MBD paralogs in soybean (GmMBD2) has subfunctionalized after their recent divergence, which was corroborated with their expression profile. Phylogenetic analysis revealed that classes of MBD proteins clustered with human MBD. Interestingly, the MBD9 may have emerged after the hexaploidization event in eudicots. We found that plants and humans share a great similarity in MBDs' binding affinity in the mCpG context. MBD2 and MBD4 from different plant species have the conserved four amino acid residues -Arg (R), Asp (D), Tyr (Y) and Arg (R)- reported to be responsible for MBD-binding in the mCpG. However, MBD8, MBD9, MBD10, and MBD11 underwent substitutions in these residues, suggesting the non-interaction in the mCpG context, but a heterochromatin association as MBD5 and MBD6 from human. This study represents the first genome-wide analysis of the MBD gene family in eurosids I - soybean and common bean. The data presented here contribute towards understanding the evolution of MBDs proteins in plants and their specific binding affinity on mCpG site.