Preparation of exine-detached pollen inNicotiana tabacum

A method of preparing exine-detached pollen inNicotiana tabacum was established. Anthers containing early-middle binucleate pollen were cold-pretreated at 4–6°C for 7–14 days, and were suspended in 0.3 mol/L sucrose solution for 2 days. During this process, the exine of most pollen grains dehisced. Then they were transferred into an enzyme solution containing 1% cellulase, 1% pectinase, 0.1% pectolyase, 1 mol/L mannitol, 0.3 mol/L sorbitol, 0.5% potassium dextran sulphate and K3 medium macro elements. After 15–20 min enzymatic maceration, the exine was detached resulting in the release of exine-detached pollen. Factors affecting preparation of exine-detached pollen were investigated, including cold-pretreatment, osmoticum concentration and enzymes used. Xia Huijun: born in Oct. 1963, Ph.D. Current research interest is in plant reproductive biology Supported by the National Natural Science Foundation of China