Varied types of pretreatments cause stress on microspore bearing anthers which induces change of the development pathway of microspore from gametophytic to sporophytic which in turn leads to the production of haploids. In our experiments different pretreatments were given to anthers of marigold before in vitro culturing of male gametophytes. Flower buds (of size 2-2.5cm) in which a majority of microspores had reached the mid- or late-uninucleate stage were tested with regard to the effect of cold pretreatment at 4°C for 3, 6, 9 and 12 days and starvation in 0.3 M mannitol solution for 2, 4, 6 and 8 days. Chilling at 4°C for 9 days resulted in highest percent of responding anthers (91.5%), highest percent caulogenesis (54.7), highest number of shoot buds per anther (7.2), highest number of regenerants per anther (5.5), lowest number of days taken for callusing (12.25) and lowest number of days taken to shoot bud induction (15.75). while starvation of anthers by using 0.3M mannitol solution for 4 days resulted in highest percent of responding anthers (81.75%), lowest number of days taken for callusing (11.75), highest percent caulogenesis (66.5%), lowest number of days taken to shoot bud induction (15.25), highest number of shoot buds per anther (7.25) and highest number of regenerants per anther (5.75). Among the two pre-treatments evaluated, cold pretreatment showed better results in terms of percent responding anthers while mannitol starvation was found best for percent caulogenesis.