Prickly acacia (Vachellia nilotica subsp. indica), one of the most harmful weeds of the Australian rangelands, has been occasionally observed displaying natural dieback symptoms since 1970's.More recently in 2010, a prominent widespread dieback event was observed among the plants growing around Richmond and Julia Creek in north-western Queensland. Affected plants were found with disease symptoms such as; ashy internal staining, defoliation, blackening of shoot tips through to widespread plant mortality. It was hypothesized that a pathogenic fungus/i could be implicated with this phenomenon and a potential for biological control of this invasive species might ensue.Fungi isolated from dieback-affected and healthy stands of prickly acacia were putatively identified by partial sequencing of the Internal Transcribed Spacer (ITS) region of genomic DNA.Botryosphaeriaceae was the best represented family of fungi associated with dieback. Among the Botryosphaeriaceae fungi, Cophinforma was found to be the most prevalent genus with 60% of the total isolates identified as Cophinforma spp. following BLAST searches. Cophinforma was also isolated from the healthy plants growing in the trial site at Richmond. Natural dieback on prickly acacia was previously observed in the surrounding areas. Phylogenetic analysis of the ITS sequences revealed the potential existence of new species of Cophinforma in Australia. Apart from Cophinforma sp., a range of other Botryosphaeriaceae and non-Botryosphaeriaceae fungi were isolated from both dieback-affected and healthy prickly acacia plants growing at different locations.Based on a seedling pathogenicity screening, a sub-set of 40 potentially pathogenic isolates across the range of species were selected to challenge the juvenile plants following a stem inoculation technique under glasshouse conditions. In the glasshouse trials, the test-isolates of Cophinforma sp.(isolated from both dieback-affected and healthy prickly acacia) and an isolate of Lasiodiplodia pseudotheobromae strain NT039 (isolated from dieback-affected Parkinsonia aculeata) consistently caused significant stem lesions, gummosis and leaf mortality. Cophinforma sp., Fusarium acuminatum, Neofusicoccum parvum and Tiarosporella graminis isolated from healthy prickly acacia also caused similar symptoms in glasshouse inoculation trials. The remaining species tested in the glasshouse trials were mostly non-pathogenic to the juvenile plants.In another study, given the observation that several fungi are often isolated from a single piece of dieback-affected stem, it was investigated whether typical dieback symptoms in prickly acacia could be replicated with combinations of different fungi. Therefore, the effect of inoculating with iii two isolates of Cophinforma sp. and one isolate of L. pseudotheobromae singly and in combination under glasshouse and field conditions was tested. In the glasshouse trial, significant stem lesions were observed with each of the test-isolates and their combinations. However, there was no signifi...