Factors Affecting Haploid Plant Production from in vitro Anther Cultures of Nicotiana Species1

Abstract

Twenty seven Nicotiana species were evaluated for their response to the in vitro culture of immature anthers on chemically defined media. Several variables were examined for their effect on haploid plant production from anther culture: stage of microspore development, anther culture medium, pretreatment of anthers prior to placing on culture medium, and stage in the flowering cycle when floral buds were collected. Thirteen Nicotiana species produced plants from anther culture. N. glutinosa L., N. knightiana Goods., N. paniculata L., N. rustica Schrank, N. sylvestris Speg. and Ccmes, and N. tabacum L. produced primarily haploid plants from anther culture. Diploid plants were obtained from the anther cultures of N. bonariensis Lehm., N. longiflora Cav., N. langsdorfii Wein., N. nudicaulis Wat., N. plumbaginifolia Viv., and N. stocktonii Brand. Plants of undetermined ploidy level were obtained from N alat Link and Otto anther cultures. Anthers cultured at the binucleate microspore stage produced plants more frequently, and more plants were produced per anther from these cultures than from anthers cultured at other microspore stages. Anthers from floral buds collected from plants in flower for an extended period did not produce as many plants as anthers from floral buds collected from plants in earlier stages of flowering. Low temperature anther pretreatment increased the number of plants produced from such anthers under certain conditions, but did not extend the number of species in which haploid plants were formed. The effect of culture medium varied with the species, length of flowering time when anthers were collected for culture, and pretreatment conditions.