Issue Archive

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

Xu Hengping, Tsao T.H.

College of Life Sciences,Peking University.Beijing 100871

Abstract

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 effeciency 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 1.2×10~(8) viable sperm cells could be obtained from 100 g fresh pollen sample within 6 h.

Key words: Zea mays L.;Inbred line;Mass isolation;Sperm cell yield

Received: 　　Accepted:

Corresponding author: 　　E-mail:

Citing This Article：

Xu Hengping, Tsao T.H.. An improved procedure for the mass isolation of viable sperm cells from pollen of Zea mays L.. Acta Physiol Sin 1997; 49 (3): (in Chinese with English abstract).