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Immunofluorescent Localization of Myosin on the Sperm Cells of Plumbago Zeylanica

Published online by Cambridge University Press:  02 July 2020

Zhaojie Zhang  and
Scott D. Russell
Zhaojie Zhang
Affiliation:
Department of Botany and Microbiology, University of Oklahoma, Norman, OK73019
Scott D. Russell
Affiliation:
Department of Botany and Microbiology, University of Oklahoma, Norman, OK73019
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Extract

Sperm cells in flowering plants are non-motile and are passive participants in their movement to the female reproductive cells. It is believed that actomyosin interaction may play a key role for sperm cell transmission in the pollen tube as well as in the embryo sac. However, indirect evidence has shown that the surface of sperm cells lacks amounts of myosin sufficient to support movement. Immunofluorescence microscopy was used in this study to further assess the presence of myosin on the surface of sperm cells of Plumbago Zeylanica.

Sperm cells of Plumbago Zeylanica were isolated according to published methods. Isolated sperm cells were blocked 20 min with 1% bovine serum albumin and 2% normal goat serum in phosphate buffered saline (PBS) (pH 7.3, in 15% sucrose), incubated 2 hrs in anti-myosin antibody (M-7648, Sigma Chemical Co., St. Louis, MO) diluted 1:10 with blocking solution, washed three times (5 min) with blocking solution and incubated 1 hr in 1:30 FITC-conjugated anti-rabbit IgG as secondary antibody (EY Labs, Inc., San Mateo, CA) in blocking solution. Samples were rinsed in PBS and mounted in an anti-fading solution with 1:1 PBS:glycerol with 3% n-propyl gallate.

Type
Developmental and Reproductive Biology
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 183 - 184
Copyright
Copyright © Microscopy Society of America 1997

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References

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6. We thank W. Strout, Greg for technical assistance. Research support through grants from the U.S. Department of Agriculture (91-37304-6471 and 95-37304-2361).Google Scholar