Occasional one-time tillage (strategic tillage, ST) is an effective tool for managing weeds and crop diseases in no-till and conversative farming systems. However, there is limited understanding of the impacts of ST on soil microbiome and their associated soil processes, particularly in dryland agriculture. This study aims to quantify the effect of one-off ST - after three years - on soil microbiomes and functions in a long-term no-till farming system under crop stubble and fertilizer management practices. The results showed that ST had marginal effects on microbial richness and diversity, enzyme activities, and catabolic function, but significantly affected the abundance of some microbial taxa (Actinobacteria, Firmicutes, Verrucomicrobia, Basidiomycota and Ascomycota) that are relevant to carbon (C) degradation. Stubble retention, regardless of tillage and fertilizer management, mainly increased the abundance of copiotrophs such as Proteobacteria (e.g., Rhizobiales) and Actinobacteria (e.g., Streptomyces and Micromonosporaceae), and affected Ascomycota and Basidiomycota. Among the management practices, stubble retention was the main factor that contributed to increased richness and diversity of the soil bacterial and fungal community. Supplementary fertilizer application, regardless of tillage and stubble management had minimal impact on bacterial and fungal richness and diversity, enzyme activity and catabolic function. The variation in bacterial community structure was influenced mainly by soil pH (c.a. 10%), while only a small but significant effect (< 7%; P = 0.001) was attributed to tillage and stubble management. Wheat grain yields ranged between 5 and 5.3 t ha-1 and were not affected by tillage, stubble, nor fertilizer management practices. Similarly, these management practices did not influence total soil C or nitrogen concentrations. Our findings show that strategic tillage, when used to address specific constraints in no-till systems in dryland agriculture, does not have a significant effect on total soil C, microbial ecology nor catabolic function.