Culture of Isolated Microspores of Saccharum spontaneum)

Microspore culture of Saccharum spontaneum L. was investigated as a means to produce haploid plants. Isolated and cultured microspores initiated sporophytic development which in 4 to 6 weeks resulted in procalli and calli.

The percentage of microspores producing multicellular procalli was increased by removing the nonviable microspores before initiating the culture, establishing an optimum population density, and using nutrient media relatively low in sucrose and NH₄NO₃.

The procalli produced from microspore culture ceased development at about the same stage as did procalli within anthers in culture. Thus, a factor or factors other than the anther wall must be responsible for the low yield of haploids from S. spontaneum.

The effect of temperature and of sucrose and 2,4-D in the media changed during the development of calli from unicellular microspores. The apparently different requirements for specific stages of development indicates that successful production of haploid plants from microspore culture will require a series of environmental and cultural manipulations.