Cultivation of grapevines in sloping soils is very widespread all over the world, representing also fundamental branch of local economy of several hilly areas. Vineyards can be managed in different ways. Agronomical practices in inter-rows can be significantly different and may influence deeply the soil properties and the grapevine root development. Therefore, this paper aimed to analyze the effects of different management practices of inter-rows. We focused on the traditional agricultural techniques of tillage and permanent grass mulching as well as the alternation of these two practices between adjacent inter-rows, in terms of: i) soil physical properties; ii) soil hydrological properties; iii) root density; iv) root mechanical properties and root reinforcement; as well as v) biodiversity. The research was conducted in several test-sites of Oltrepò Pavese (Lombardy region, north-western Italy), one of the most important Italian zones for wine production in northern Italian Apennines. Among the examined soil properties, hydraulic conductivity was the most influenced soil property by different soil management practices. The absence of soil tillage allowed to increase superficial (first 0.2 m of soil) hydraulic conductivity, as a consequence of higher macroporosity and amount in organic matter. Vineyards with alternation management (grass mulching together with tillage) of inter-rows had the highest root density and the strongest root reinforcement, of up to 45% in comparison to permanent grass cover, and up to 67-73% in comparison to tilled vineyards. Soil microarthropod communities had more complexity where sustainable agricultural practices (permanent grass cover; alternation management of the inter-rows) were applied. The results of this study yielded important information to establish effective management practices of vineyards such as conserving organic matter and reducing slope instabilities by a better development of root apparatus in the soil.