A field experiment was conducted to evaluate different cut flower cultivars under poly-tunnel conditions, focusing on various growth and floral characteristics of cut flowers at the Horticulture and Forestry Research Station, Tamil Nadu Agricultural University (TNAU), Kodaikanal, from 2022 to 2025 to assess the impact of protected cultivation on the performance of Lisianthus (Eustoma grandiflorum), Asiatic lily (Lilium asiatica), and gerbera (Gerbera jamesonii). In gerbera (2022–2023), the polytunnel environment consistently outperformed other structures, recording the highest number of leaves (18.07), leaf length (40.54 cm), leaf breadth (15.14 cm), plant spread (60.08 cm NS × 55.31 cm EW), and earliest flowering (53.73 days to bud visibility). Maximum stalk length (77.22 cm), flower diameter (14.10 cm), and ray floret count (60.73) were also observed in the polytunnel, alongside the highest chlorophyll a (11.57 mg/g), chlorophyll b (4.2 mg/g), total chlorophyll (15.26 mg/g), and soluble protein content (85.03 mg/g). Among cultivars, Haimi exhibited superior growth, flowering, and yield (2.27 flowers/plant), while Breakdance excelled under shade net conditions. For Asiatic lily (2023–2024), polytunnel-grown plants achieved the tallest height (107.06 cm), highest leaf count (77.93), longest leaf length (12.54 cm), and earliest bud visibility (47.59 days). Cultivar Tirreno (V5) attained the maximum plant height (108.25 cm), whereas Cavalia (V2) recorded the largest flower diameter (18.46 cm), the highest number of florets per spike (4.53), and maximum chlorophyll a content (12.10 mg/g). Polytunnel-grown lilies also exhibited the longest vase life (16.27 days), highlighting the benefits of protected cultivation for commercial floriculture. In Lisianthus (2024–2025), cultivar Rosita 3 Clear Pink (V1) showed the highest plant height (106.33 cm), leaf number (60.40), longest internodal length (8.82 cm), earliest flower bud initiation (91.18 days), and shortest time from bud initiation to flower opening (24.59 days). Cultivar Aube 4 Pure White (V5) excelled in floral traits, recording the highest shoots per plant (4.84), flowers per stem (10.70), petal count (31.34), and maximum bud length (4.82 cm), flower length (6.25 cm), and flower diameter (9.74 cm), indicating its suitability for producing high-quality blooms. Overall, these studies demonstrate the significant advantages of protected cultivation, particularly polytunnel structures, in enhancing the vegetative growth, flowering, yield, and physiological traits of Lisianthus, Asiatic lily, and gerbera. Cultivars such as Aube 4 Pure White, Rosita 3 Bright Blue (Lisianthus), Cavalia and Tirreno (Asiatic lily), and Haimi and Breakdance (gerbera) exhibited superior performance, offering valuable insights for optimizing floriculture production under cool, humid conditions of Kodaikanal and similar agro-climatic zones.