On Plant Responses to D-Amino Acids Features of Growth, Root Behaviour and Selection for Plant Transformation

Amino acids have been regarded as potential plant nitrogen sources for more than a century. Some amino acids have been shown to support growth while others have growth-retarding effects. The D-isomers with notably adverse effects on plants’ growth and development include D-serine and D-alanine. Recently, D-serine has been recognised as an endogenous ligand of receptor channels mediating calcium fluxes in plants, but otherwise little is known about endogenous roles of D-amino acids in plants. In the studies underlying this thesis, the negative responses to D-serine and D-alanine were converted to positive effects in Arabidopsis thaliana (Arabidopsis) plants by introducing either of two D-amino acid-metabolising enzymes. Transgenic Arabidopsis lines expressing either the D-serine dehydratase (dsdA) gene from Escherichia coli or the D-amino acid oxidase (DAO1) gene from Rhodotorula gracilis grew with otherwise toxic D-amino acids as the sole nitrogen source. I also expressed a transporter specific for D-amino acids, which further increased the transgenic plants’ growth with D-serine as sole nitrogen source. Hence, both assimilation and uptake restrictions can limit plant growth on D-amino acids. The growth of transgenic lines on D-serine or D-alanine provides an unambiguous and highly visible phenotype, which is essential for a selectable marker. Thus, expressing of either the dsdA or DAO1 genes generated transformants that are easy to screen. Furthermore, the DAO1 gene can be readily used for either positive or negative selection, depending on the substrate, thus it provides a unique conditional, substrate-dependent positive/negative selectable marker for plant transformation. In summary, the presented work demonstrates that introducing the ability to catalyse a single metabolic step can allow plants to exploit an otherwise inaccessible or toxic form of organic nitrogen, and provides a versatile marker based on nitrogen nutrition for selecting transgenic plants. A possible role for D-serine in plants’ touch response is also reviewed in the thesis.