Fluorescence microscopy coupled with biochemical analysis has helped resolve the differential activity of the enzyme, β-l, 3- glucanase that catalyses the dissolution of callose. The enzyme activity is relatively low during the first meiotic division but reaches its peak at the time of spore release. This sequence of events controlling the enzyme activity is important, as an early dissolution of callose may lead to sterility. Use of autoradiographic and fluorochromatic techniques demonstrated the reduced permeability of the callose wall and the resultant chemical isolation of microspores.
A combination of phase-contrast and fluorescence microscopy also revealed that the callose wall during exins formation serves as a mold or template. Factors that trigger the onset and regulation of meidsis in anther and ovule can be worked out by employing labeled reagents. Biochemical studies through isoenzyme markers have already demonstrated specific stages of meiotic process. There is an increase in the activity of acid phosphatase at zygotene, followed by subsequent decline. At pachytene, marked activation of endonucleases is recorded. Isoperoxidases serve as markers in normal pollen development in Mercurialis annua.