There is little available information on the effects of temperature and CO 2 enrichment on stomata anatomical characteristics of plants. Effect of these two microclimates was studied on five rose (Rosa spp.) cultivars, viz. ‘First Red’ (used as check), ‘Arjun’, ‘Raktima’, ‘Raktagandha’ and ‘Pusa Pitamber’. Budded, single-stemmed rose cultivars having five lateral buds were grown in controlled environment growth cabinets under enriched CO 2 (1000μmolmol −1 ) and optimum (28/18°C, T 0 ) or high (35/25°C, T 1 ) temperature for 50 days. All observations were made on the abaxial leaf surface. Significant increases in stomatal density (68.7%), index (29.6%) and epidermal cell density (37.3%) were recorded in plants grown at high temperature over control with CO 2 enrichment. The cultivars responded differently in terms of length and width of guard cell and stoma (pore) under high temperature, however, the values averaged over treatments showed a significant reduction in these parameters. Further, number of stomata per leaf was higher (28.3%) in plants grown at high temperature, except First Red. A reduction in mean leaf area (26.7%) and dry mass (32.0%) was recorded at high rather than optimum temperature. The specific leaf area was maximum in Arjun (87%) while in First Red, a 14% reduction was noted at high temperature.The changes in the stomatal aperture (length and width of guard cell and stoma) did not show a clear pattern in response to higher temperature under CO 2 enrichment, which suggests that it is a genotypic character. Our data also show that temperature is potentially a more important factor determining stomatal anatomy than other environmental factors. Thus, this study provides an important database on stomatal anatomy for two important microclimates which is of direct significance to ornamental horticulture.