title: Arabidopsis aldehyde dehydrogenase 10 family members confer salt tolerance through putrescine-derived 4-aminobutyrate (GABA) production.
abstract: Polyamines represent a potential source of 4-aminobutyrate (GABA) in plants exposed to abiotic stress. Terminal catabolism of putrescine in Arabidopsis thaliana involves amine oxidase and the production of 4-aminobutanal, which is a substrate for NAD+-dependent aminoaldehyde dehydrogenase (AMADH). Here, two AMADH homologs were chosen (AtALDH10A8 and AtALDH10A9) as candidates for encoding 4-aminobutanal dehydrogenase activity for GABA synthesis. The two genes were cloned and soluble recombinant proteins were produced in Escherichia coli. The pH optima for activity and catalytic efficiency of recombinant AtALDH10A8 with 3-aminopropanal as substrate was 10.5 and 8.5, respectively, whereas the optima for AtALDH10A9 were approximately 9.5. Maximal activity and catalytic efficiency were obtained with NAD+and 3-aminopropanal, followed by 4-aminobutanal; negligible activity was obtained with betaine aldehyde. NAD+reduction was accompanied by the production of GABA and β-alanine, respectively, with 4-aminobutanal and 3-aminopropanal as substrates. Transient co-expression systems using Arabidopsis cell suspension protoplasts or onion epidermal cells and several organelle markers revealed that AtALDH10A9 was peroxisomal, but AtALDH10A8 was cytosolic, although the N-terminal 140 amino acid sequence of AtALDH10A8 localized to the plastid. Root growth of single loss-of-function mutants was more sensitive to salinity than wild-type plants, and this was accompanied by reduced GABA accumulation.
authors.affiliation: Department of Plant Agriculture, University of Guelph, Guelph, Ontario, N1G 2W, Canada.
authors.forename: Adel
authors.initials: A
authors.lastname: Zarei
authors.affiliation: Department of Plant Agriculture, University of Guelph, Guelph, Ontario, N1G 2W, Canada.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Trobacher
authors.affiliation: Department of Plant Agriculture, University of Guelph, Guelph, Ontario, N1G 2W, Canada.
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Shelp
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics / Q000502:physiology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D019697:Onions; D010084:Oxidation-Reduction; D011700:Putrescine / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D055049:Salt Tolerance; D012492:Salts / Q000378:metabolism; D013379:Substrate Specificity; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D011994:Recombinant Proteins; D012492:Salts; D005680:gamma-Aminobutyric Acid; D000444:Aldehyde Dehydrogenase; D011700:Putrescine