A Fluorescent Brightener used for Pollen Tube IdentificationIn Vivo

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Abstract
Squash preparations of styles stained in watersoluble aniline blue and viewed under ultra-violet illumination are regularly used for examining pollen tubes because the callose plugs fluoresce brightly under these conditions. Tubes are therefore clearly distinguish from the astylar tissue and may be readily counted and measured. This method has proved to be quite unsatisfactory for plum pollen tubes, since they contain very few cause plugs and better results have been obtained with a mixed stain of 0.1% aniline blue and 0.07% of the fluorescent brightener ‘Calcofluor White M2R New’. Styles are softened by autoclaving in 50 g/1 sodium sulphite, rinsed and stained for ten minutes, then squashed and examined with a fluorescence microscope in the usual way. Callose deposits, when present, fluoresce bright yellow, but lengths of tube with no deposits can also be clearly identified and followed, permitting easier, faster and more accurate assessments of pollen tube length and numbers in plum and pear styles.