Background Acupuncture services form a significant part of the Australian healthcare setting, with national registration of acupuncture practitioners, public subsidies for acupuncture services and high use of acupuncture by the Australian public. Despite these circumstances, there has been little exploration of the interface between acupuncture providers and conventional primary healthcare practitioners in rural and regional Australia. Methods A 27-item questionnaire was sent by post in the second half of 2010 to all 1486 general practitioners (GPs) currently practising in rural and regional Divisions of General Practice in New South Wales, Australia to explore their practices and attitudes to a variety of complementary and alternative medicine (CAM) practices. Their responses on other therapies have been published previously; this report covers acupuncture. Results A total of 585 GPs completed the questionnaire; 49 were returned as ‘no longer at this address’, resulting in an adjusted response rate of 40.7%. Two-thirds of GPs (68.3%) referred patients to an acupuncturist at least a few times per year, while only 8.4% stated that they would not refer patients to an acupuncturist under any circumstances. GPs being older (OR=6.08), GPs being women (OR=2.94), GPs practising in a rural rather than remote area (OR=6.25), GPs having higher levels of self-reported knowledge of acupuncture (OR=5.54), the use of complementary medicine (CAM) by a GP for their personal health (OR=2.37), previous prescription of CAM to other patients (OR=2.99), lack of other treatment options (OR=4.31) and GPs using CAM practitioners as the major source of their CAM information (OR=3.05) were all predictive of increased referral to acupuncture among rural GPs. Conclusions There is a significant interface between acupuncture and Australian rural and regional general practice, with generally high levels of support for acupuncture.