Production of haploid tobacco plants using anther culture
Haploids are plants that have the gametic, or n, number of chromosomes. They are very useful in genetic and breeding studies because recessive genotypes are easily identified. For many years, few haploids were available to geneticists and breeders. This situation began to change in the 1960s with the discovery that haploids of Datura species and tobacco could be produced by culturing anthers. This chapter illustrates the technique of anther culture using tobacco. It determines the optimum stage of microspore development for anther culture. The chapter then discusses the use of cold pretreatments for enhancing haploid production from anther culture. Cultures can be scored for number of haploids produced approximately eight weeks after culture initiation. Using fine-tipped forceps gently separate plants in order to get accurate counts. Unless these plants are to be used for other experiments, cultures can be scored outside of aseptic conditions.