A comparative analysis of five species of Cucurbitaceae members such as Lagenaria vulgaris, Benincasa hispida, Cucurbita maxima, Luffa acutangula and Cucumis sativus was carried out on the powder microscopic observation, petiole and leaf anatomy. Similarities were observed in the distribution, differentiation and number of layers of cells and tissues in the petiole and leaf anatomical sections. However, variations were observed in the numbers of vascular bundles in the petiole and leaf and they were bicollateral in all the species. Numbers of petiolar vascular bundles were 20 in Lagenaria vulgaris, 10 in Benincasa hispida, 12 in Cucurbita maxima and 9 in Cucumis sativus and Luffa acutangula respectively. Presences of stomata are commonly seen in all five species. The Trichomes were multicellular, glandular or non-glandular, unbranched and uniseriate. Non-glandular trichomes were predominantly seen in the genera studied, whereas Lagenaria vulgaris and Luffa acutangula have glandular trichomes. The variations in the number of tiers within these trichomes were taxonomically significance.