Elucidating the Role of N-Acylethanolamine/Anandamide Metabolism in the Moss Physcomitrella Patens

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In plants, saturated and unsaturated N-Acylethanolamines (NAEs) with acyl chains 12C to 18C are reported for their differential levels in various tissues and species. While NAEs were shown to play a vital role in mammalian neurological and physiological functions, its metabolism and functional implications in plants however, remains incomplete. Recently, anandamide (NAE 20:4), an essential fatty acid neurotransmitter in mammalian system, was identified in moss Physcomitrella patens, in addition to other types of NAEs. Bryophytes display high tolerance to abiotic stress and thus presence of anandamide in moss, but not in higher plants, suggests that NAE 20:4 might have contributed to their survival in harsh environmental conditions. Therefore, we hypothesize the anandamide metabolic pathway might play a role in mediating stress responses in P. patens. To this extent, using previously identified NAE-metabolic genes in mouse and/or Arabidopsis as templates, we identified moss orthologs for enzymes that likely participate in anandamide metabolism. We identified members of metallo-hydrolase superfamily and a/Β-hydrolase4, and five putative fatty acid amide hydrolases, which may hydrolyze N-acylphosphatidylethanolamine and NAE, respectively. Electronic fluorescent pictograph analyses of these orthologs in moss revealed differential developmental stage-specific expression patterns in gametophyte and sporophyte stages. We are currently examining expression pattern for these putative NAE-metabolic pathway genes, along with anandamide levels, in different tissues and developmental stages of moss subjected to water stress in the presence of anandamide. These transcript and metabolite levels in moss subjected to stress are expected to offer better understanding of the role of anandamide in mediating stress responses and further allow us to identify candidate genes that might participate in NAE metabolism. Our studies are aimed at functional validation of candidate genes and generating moss transgenic lines with altered NAE metabolite profile. Our long-term goal is to conduct comprehensive analyses of NAE metabolite mutants to determine their role in growth and development, and mediating stress responses in plants.


Johnson City, TN

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