Characterization of LYM Genes in Medicago Truncatula

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Characterization of LYM genes in Medicago truncatula. M. truncatula contains a large family of genes encoding LysM-domain containing receptor-like proteins (Arrighi et al., 2006). In addition, several proteins containing single or multiple LysM domains can be predicted from available sequencing resources. Among these, two transcripts encoding proteins with structural homology to the chitin elicitor-binding protein CEBiP from rice (Oryza sativa (Kaku et al., 2006)) were identified, MtLYM1 and MtLYM2 (Arrighi et al., 2006). The proteins deduced from these transcripts featured signal peptides, indicating an extracellular localization (Fig. 1). Moreover, both proteins were predicted to be modified by glycosylphosphatidylinositol (GPI) for anchorage at the cell surface after removal of the C-terminal transmembrane domain. The genes encoding MtLYM1 and 2 contained five exons/four introns and four exons/three introns, respectively (Fig. 1), from which only one intron position was conserved between both genes. Interestingly, a small intron was present in MtLYM2 inside of the region encoding the three LysM domains, which was never observed for other genes encoding LysM-containing proteins in M. truncatula (Arrighi et al., 2006) We analyzed the phylogeny of LYMs by performing a consensus search of fully sequenced plant genomes for encoded proteins with similarity to LYMs from M. truncatula. Proteins, which were not predicted to contain a signal peptide or which were shorter than 300 residues were removed to obtain a list of 53 LYMs (suppl Table xx). In contrast to LYPs, which are defined, less stringently, as LysM-containing extracellular proteins and were detected in the moss Physcomitrella patens (Zhang et al., 2009), LYMs are present ... ... middle of paper ... ...n). Strongest promoter activity was observed in root tips and in lateral root emergences (Fig. 7A-D). Expression level and distribution was rather variable, with occasional staining of root hairs (Fig. 7A, D, F). During infection with Sinorhizobium meliloti (pXLGD4), GUS activity was detectable in developing nodules (Fig. 7E, F). Major nodules showed little GUS activity in the outer layers (Fig. 7G, H). Sectioning of old nodules (4 weeks pi ??) prior to GUS staining uncovered a peculiar expression pattern of MtLYM2. Here, promoter activity showed to be restricted to vascular bundles only, with, in some cases, very little staining of the outer cortex (Fig. 7x). In summary, the histochemical localization of GUS activity in proLYM2::GUS-int expressing roots and nodules confirmed our earlier observation of reduced transcript and protein levels in nodules versus roots.

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