An investigation on air layering in guava as influenced by growth regulators and Azospirillum was carried out in the mother plants block of guava at the Silver Jubilee Orchard (SJO), Department of Horticulture, College of Agriculture, University of Agricultural Sciences, Dharwad during 2010-11. The study aimed to find out suitable root inducing treatment and its optimum concentration for maximum survival percentage in mature shoot air-layers of guava Cv. L-49. There are 14 treatment combinations with three replications laid out in Randomised Block Design. In general, both growth regulators viz. IBA, NAA alone and in combination with Azospirillum favoured rooting in air-layers. A synergestic effect was noticed in terms of higher percentage of rooting where the two growth regulators were used in combination with Azospirillum. Among the different combinations, the layers which had received Azospirillum 37.5g + IBA (Indole butyric acid) 3000 ppm + NAA (Naphthalene acetic acid) 3000 ppm (T12) recorded significantly higher percentage (91.68%) of rooting with desirable root characters such as higher number of primary and secondary roots, longer length of primary roots and higher girth of primary roots. Next to this treatment, the other favourable treatments were IBA, 3000 ppm + NAA, 3000 ppm (T6), IBA, 2000 ppm (T1), Azospirillum 37.5g + IBA, 4000 ppm (T8), Azospirillum 37.5 g + IBA, 6000 ppm (T9), Azospirillum 37.5 g + IBA, 2000 ppm + NAA, 2000 ppm (T11). The use of medium concentration (3000 ppm) of both the growth regulators (IBA and NAA) with Azospirillum 37.5g was more effective as compared to either lower (2000 ppm) or higher (6000 pm) concentrations of IBA and NAA. With regard to survivability of rooted layers, maximum survival percentage (98.14%, 60 days after separation) was noted in the layers treated with Azospirillum 37.5g + IBA, 3000 ppm + NAA, 3000 ppm (T12).