Use of Pectinase in Pollen Tube and Pollen Mother Cell Smears

Styles of Physalis pruinosa L. and P. ixocarpa Brot. were fixed in a modification of Carnoy's fixative (alcohol, chloroform, acetic acid; 3:2:1) 48 hr after pollination. After 24 hr, the styles were transferred to a 5% aqueous pectinase solution (adjusted to pH 4) on a spot plate for 24 hr. The styles were then transferred with a brush into another spot containing a mixture of 15 parts of 1% aqueous light green to 1 part Darlington and LaCour's acid fuchsin light green stain. The styles were mounted individually in glycerol after 24 hr of staining. Generally the weight of the cover slip was enough to adequately flatten the style. The unstained glassy pollen tubes were easily discernible throughout their lengths on a background of the purplish blue stylar tissues. It was also found that by putting fresh or fixed anthers into the pectinase solution for about an hour and then macerating them in acetocarmine, almost all the pollen mother cells were separated from each other. This increased the number of well flattened cytologically desirable cells.