SummaryThe Egyptian clover or berseem (Trifolium alexandrinum L.) is widely cultivated as a winter season forage crop in about two million hectare of land in India. It is widely accepted because of its multicut nature, high yield and nutritional value. A plausible approach for increasing yield could be through increasing photosynthetically active leaf area and consequently the biomass. The present investigation deals with induction of polyploidy and evaluation of tetraploids vis-à-vis diploids lines of Egyptian clover for multifoliate leaf formation. Immersing of pre-hydrated seeds, from diploid multifoliate plants, in colchicine followed with seedlings immersed in 0.1 and 0.2% colchicine solution for 24 and 48 h, respectively, was effective in inducing tetraploidy. Distinct characteristic features observed among induced pentafoliate tetraploid plants were presence of serrate leaflet margin, prominent rachis and bold seeds. Leaves were thick, succulent, and hairy with apical notch characterized by presence of pigmentation on the outer margin. The autotetraploids had better expression of pentafoliate trait than in diploid plants.Key words Colchicine, Multifoliate, Pentafoliate, Polyploidy, Tetraploid, T. alexandrinum, Egyptian clover, Fodder.The Egyptian clover or berseem (Trifolium alexandrinum L.) is a diploid species (2n=16) commonly cultivated as a winter annual fodder crop in tropical and subtropical countries. It is cultivated as a winter season forage crop in about two million hectare of land in India. It is widely accepted because of its multicut nature, high yield and nutritional value. In the past, efforts have been made to induce genetic variability through mutation, polyploidization and selection but the results were not very encouraging. A plausible approach for increasing yield could be through increasing photosynthetic area, i.e. leaf area, which will help in increasing the photosynthetically active leaf area and consequently the biomass yield. The number of leaves per plant and leaflets per leaf in the Egyptian clover contribute to higher leaf-stem ratio (Bakheit 1996). Cultivars of Egyptian clover possess trifoliate leaves in general. However, occasional occurrence of multifoliate plants in natural population is reported (Shukla and Malaviya 1986). The frequency of multifoliate plants in the natural population of berseem was 0.004% only. Such multifoliate plants possessed only 1 to 2% multifoliate leaves. Likewise, white clover occasionally produces a leaf with four (or more) leaflets. The genetics underlying this phenomenon has been partly worked out (Tashiro et al. 2010). One such reference to multifoliolate leaves in white clover (Trifolium repens L.) is in the registration of FL-ML white clover germplasm (Baltensperger et al. 1991). Knight (1969) in Trifolium incarnatum and Jaranowski and Broda (1978) in Trifolium pratense also reported multifoliate leaf formation. However, there is no report of developing pure pentafoliate in any