Isolation of Viable Wheat Male Gametophytes of Different Stages of Development and Variations in Their Protein Patterns

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Abstract
Procedures have been designed to isolate viable immature male gametophytes from wheat (Triticum aestivum L.) anthers of different stages of development. Maceration of anthers by a micro-blender allowed for the release of numberous intact vacuolate microspores. Tris-buffered media prevented tricellular pollen grains from bursting during the isolation procedure. Proteins from the undamaged male gametophytes have been analyzed by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A set of new proteins was detected at the onset of the second pollen grain mitosis. The results demonstrate the opportunity for studies on haploid gene expression at the translational level.