Selection and microculture of single embryogenic cell clusters in Japaneseconifers: Picea jezoensis, Larix leptolepis and Cryptomeria japonica

A microculture method for single embryogenic cell clusters was established for several Japanese conifer species, namely Picea jezoensis, Larix leptole pis and Cryptomeria japonica. Individual small, dense cell clusters were id entified and picked up by a micromanipulator and cultured in 50 mu l liquid medium in a 96-well culture plate. In an three species studied, a majority of the cell clusters actively proliferated within the wells. Maturation of somatic embryos was successful when the newly proliferated cell clusters w ere transferred to solid abscisic acid-containing medium. Thus, the small, dense cell clusters could be a useful morphological. marker for cells capab le of proliferation and differentiation.