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An improved procedure for the mass isolation of viable sperm cells from pollen of Zea mays L

An improved procedure for the mass isolation of viable sperm cells from pollen of Zea mays L

Acta Phytophysiologica Sinica 23(3): 245-250

Mass isolation of viable sperm cells from maize pollen was further studied to increase the yield of sperm cells. Four different approaches on pollen bursting and sperm collection were tried. Results showed that pollen bursting rate could reach as high as 95% with BKS35 preincubation followed by buffered osmotic shock solution (in sucrose 0.44 mol/L, Mes 2 mmol/L and 0.1% BSA, pH 6.7). After examining the isolation efficiency of 4 different Percoll solutions, it is found that the higher sperm yield (ca. 23.4%) could be obtained using 30% Percoll in buffered sucrose solution. Using the above modified two-step osmotic shock followed by 30% Percoll centrifugation, about 1.2 X 108 viable sperm cells could be obtained from 100 g fresh pollen sample within 6 h.

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