Microspore culture protocol for Indonesian Brassica oleracea

A microspore culture protocol for Brassica oleracea of Indonesian origin (cv. 'Kemeh') has been successfully established. A high number of embryos formed with high microspore density i.e. 15 x 10(4) cells/ml. Embryo formation was improved by using flower buds (4.5-4.6 mm in length) as explants, a temperature treatment at 30.5 degrees C for 48 h and then transfer to 25 degrees C continuously until embryos formed. A total of 295 embryos were obtained from 189 buds, 30% of which were abnormal (i.e. with an abnormal cotyledon or lacking hypocotyls). All normal embryos that grew and survived, 165 in total, were successfully transferred to soil and grew well in plastic bags (15 cm in diameter) containing a mixture of burned-rice husk and organic manure (1:1, v/v).