Putative dicarboxylate and amino acid transporters in soybean (Glycine max L.): a molecular characterisation

Helle Martha Christophersen

Research output: ThesisDoctoral Thesis

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[Truncated abstract] Some plants, such as legumes, are able to use atmospheric nitrogen as a nitrogen source due to the nitrogen-fixing bacteria residing in specialised root structures called nodules. The exchange of carbon and nitrogen between the host plant (legume) and the nitrogen- fixing micro-symbiont is vital for biological nitrogen fixation. In particular, transport of C4-dicarboxylates, mainly malate, from the plant to the micro-symbiont, and the reverse transport of fixed nitrogen in the form of ammonium are essential for symbiotic nitrogen fixation. In the legume nodule, the symbiosome membrane (SM) surrounds the bacteroid and all exchanges of metabolites and nutrients that occur between the plant and the micro-symbiont must cross this membrane. Recently it has been established that cycling of amino acids across the SM is also critical for optimal symbiotic nitrogen fixation. Therefore to fully understand this agriculturally significant phenomenon, the mechanisms facilitating these exchanges need to be investigated. The major aim of this study was to increase the understanding of nutrient exchange within the nodule at the molecular level by isolating and characterising genes encoding transporters responsible for malate and amino acids transport in soybean (Glycine max, L.), with particular interest in genes significantly or highly expressed in nodules. A combination of molecular and biochemical techniques was used to achieve this. ... Southern blot analysis showed that a small gene family of up to five members encodes these proteins in soybean. A full-length cDNA, designated GmAAP5, was isolated that encodes a novel, putative amino acid transporter. Molecular characterisation of this cDNA and that of GmAAP1 (GenBank Accession no: AY029352), a previously identified putative amino acid transporter gene, was done. Expression analyses showed relatively high expression of GmAAP5 in soybean nodules compared to that in leaf and root tissues, while GmAAP1 showed uniformly high expression in root, leaf and nodule tissues. Phylogenetic analysis of the deduced amino acid sequences of known functional AAPs from dicotyledonous plants revealed that GmAAP1 is most closely related to AAP2 from V. faba, while GmAAP5 is more closely related to AAPs from non-leguminous plants than from leguminous plants. Based on the functional characterisation of the AAPs with which GmAAP1 and GmAAP5 cluster, it is likely that both transporters are neutral and acidic amino acid transporters within the AAP subfamily.
Original languageEnglish
QualificationDoctor of Philosophy
Publication statusUnpublished - 2006


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