Withania coagulans is a valuable medicinal plant with high demand, but its wild growth and local usage pose a threat to its natural habitat. This study aims to understand the plant's growth, anatomy, and physiology in different environmental conditions to aid in conservation and re-vegetation efforts. Fifteen differently adapted populations of Withania coagulans were collected from diverse ecological regions, viz., (i) along the roadside, (ii) hilly areas, (iii) barren land, and (iv) wasteland to unravel the adaptive mechanisms that are responsible for their ecological success across heterogenic environments of Punjab, Pakistan. The roadside populations had high values of photosynthetic pigments, total soluble proteins, root endodermis thickness, stem and leaf cortical thickness, and its cell area. The populations growing in hilly areas showed better growth performance such as vigorous growth and biomass production. Additionally, there was enhanced accumulation of organic osmolytes (glycine betaine and proline), chlorophyll content (chl a/b), and enlarged epidermal cells, cortical cells, vascular bundles, metaxylem vessels, and phloem region in roots. In case of stem area, epidermal thickness, cortical thickness, vascular bundle, and pith area showed improved growth. However, the barren land population showed significant increase in carotenoid contents, vascular bundle area, and metaxylem area in roots, and xylem vessels and phloem area in stems and leaves. The wasteland population surpassed the rest of the populations in having greater root dry weight, higher shoot ionic contents, increased root area, thick cortical, and vascular bundle area in roots. Likewise, cortical thickness and its cell area, and pith area in stems, whereas large vascular bundles, phloem region, and high stomatal density were recorded in leaves. Subsequently, natural populations showed the utmost behavior related to tissue organization and physiology in response to varied environmental conditions that would increase the distribution and survival of species.