Biological nitrogen fixation by two Acacia species and associated root-nodule bacteria in a suburban Australian forest subjected to prescribed burning

Purpose Prescribed burning is a forest management practice which can lead to nitrogen (N)-limited conditions. This study aimed to explore whether biological N 2 fixation (BNF) remained the main source of N acquisition for two understorey Acacia species in a Eucalyptus -dominated suburban forest of subtropical Australia, 3 to 6 years after prescribed burning. Root-nodule bacteria associated with these acacias were also characterised to unravel the differences in rhizobial communities between sites and species. Material and methods Two sites, burned 3 and 6 years before sample collection, were selected within a dry subtropical forest of south-east Queensland, Australia. Leaves were collected from individuals of Acacia disparrima and A. leiocalyx at each site to determine leaf total carbon (C) and N content, C and N isotope composition (δ 13 C and δ 15 N) and the percentage of N derived from atmospheric N 2 . Nodules were harvested from both acacia species at each site to isolate root nodule bacteria. Bacterial isolates were processed for 16S rDNA gene sequencing. Results and discussion Generally, no differences were found in plant physiological variables between the two acacia species. Six years after the fire, both species still depended upon BNF for their N supply, with a higher dependence in winter than in summer. Fire, although of low intensity, was likely to have created a N-limited environment which induced the reliance of legumes on BNF. Root nodule bacteria were dominated by non-rhizobial endophytes, mainly from the Firmicutes phylum. No difference in nodule bacterial diversity was found between sites. The relative abundance of rhizobial genera varied amongst plant species and sites, with a shift in dominance from Bradyrhizobium to Rhizobium species between sites 1 and 2. Conclusions Our results show that even 6 years after burning, ecosystem remained under N stress and BNF was still the main mechanism for N acquisition by the understorey legumes.