ESTABLISHMENT AND GROWTH CHARACTERIZATION OF SUSPENSION CULTURES OF CELLS FROM BARBULA UNGUICULATA

Abstract

  We established suspension cultures of spherical cells from the moss, Barbula unguiculata. The cells remain in an undifferentiated callus state after repeated subcultures for more than two years on a solid or in a liquid medium, which is a modification of Murashige-Skoog's medium without plant hormones. Two stable cell lines were obtained, one of which utilizes either ammonia or nitrate (NA-cells) and the other utilizes only ammonia (AS-cells). The cells of the two lines grew actively in the light with concomitant synthesis of chlorophyll, but their growth required the presence of glucose in the culture media. Their growth in the light was only partly inhibited by DCMU (dichlorophenyldimethylurea) in the concentration which completely inhibited photosynthetic oxygen evolution. Both NA- and AS-cells required the presence of an organic acid, such as that in the Krebs cycle, or the presence of CaCO₃, for their growth on ammonia, but the growth of NA-cells on nitrate did not require such additions. Cells of the two lines could grow and increase their mass in the dark with definite synthesis of chlorophyll, but protonema-formation did occur during growth in the dark.