After purifying plasma membranes from isolated maize sperm cells by aqueous polymer two-phase partition, peripheral and integral proteins were solubilized from the plasma membrane with Triton X-114 and separated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Silver staining revealed 10 bands (19-68 kDa) of peripheral membrane proteins and about 40 bands (12-120 kDa) of integral proteins. Peroxidase-conjugated Con A was used to detect the surface glycopeptides. It was found that Con A particularly stained 8 peripheral polypeptide bands, including 68, 66, 55, 51,48, 44, 36, and 32 kDa, and 6 integral polypeptide bands, 68, 51, 48, 44, 38, and 34 kDa. These bands differed from those of somatic samples. Staining specificity was demonstrated by the control in the presence of competing inhibitory sugar. The above result indicates the existence of mannosyl and glucosyl residues in the surface glycoproteins of maize sperm cells. The prominent peripheral 68 kDa polypeptide was further separated into 4 spots by isoelectric focusing and sodium dodecyl sulfate two-dimensional (IEF-SDS 2-D) electrophoresis, showing pI values from 5.5 to 5.8. Three prominent glycopeptides (68, 48, and 32 kDa) were localized on the plasma membrane of maize sperm cells via the fiuorescein isothiocyanate (FITC) technique. About 25% of sperm cells showed an intense positive reaction in each immunological labelling. The results agree with our previous labelling of the surface of isolated viable maize sperm cells with Con A-FITC.
Estrogens, a group of steroidal sex hormones, were determined in pollen, styles or whole pistils using a highly sensitive and specific radioimmunoassay (RIA) and found to be present at levels between 10-11 and 10-lo g/g FW. In the style of Lilium davidii Duch., the levels of total estrogen decreased after self-pollination of the open flower. After bud pollination, which partly overcame incompatibility, the levels of both estradiol and total estrogen increased. The levels of estradiol and total estrogen in the pistils of Brassica pekinensis Rupr. increased after compatible pollination during the first 3 days following pollination and decreased thereafter when the data were expressed on a FW basis. These results suggest that estrogens are to some extent associated with the reproductive processes.
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