Hostname: page-component-5c6d5d7d68-wpx84 Total loading time: 0 Render date: 2024-08-18T10:18:45.233Z Has data issue: false hasContentIssue false

Isolation of viable sperm cells from tobacco (Nicotiana tabacum)

Published online by Cambridge University Press:  26 September 2008

Yajuan Cao
Affiliation:
Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma, USA.
Adonis Reece
Affiliation:
Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma, USA.
Scott D. Russell*
Affiliation:
Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma, USA.
*
Scott D. Russell, Deparment of Botany and Microbiology, University of Oklahoma, Norman, OK 73019-0245, USA. Telephone: +1-405-325-6234. Fax: +1-405-325-7619. e-mail: srussell@ou.edu.

Summary

Viable sperm cells of Nicotiana tabacum were isolated by the semi-vivo technique. After pollination, excised styles were floated, cut end immersed, in a solution of 15% sucrose with 0.01% boric and 0.03% Ca(NO3)2 at 27°C in a growth chamber until pollen tubes emerged. After sperm cells were formed (at least 8 h after pollination) tubes were immersed in a 9% mannitol solution. In this solution, sperm cells are nearly ellipsoidal and retain viability for over 6 h.

Type
Short communication
Copyright
Copyright © Cambridge University Press 1996

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Cass, D.D. (1973). An ultrastructural and Nomarski-interference study of the sperms of barley Can. J. Bot. 51, 601–5.CrossRefGoogle Scholar
Faure, J.-E., Digonnet, C. & Dumas, C. (1994). An in vitro system for adhesion and fusion of maize gametes. Science 263, 1598–600.CrossRefGoogle Scholar
Hiratsuka, S., Takahashi, E. & Hirata, N. (1982). Pollen tube growth in detached styles of japanese pear, Pyrus serotina REHD. J. Palynol. 18, 133–19.Google Scholar
Keijzer, C.J., Reinders, M.C. & Leferink-ten Klooster, H.B. (1988). A micromanipulation method for artificial fertilization in Torenia. In Sexual Reproduction in Higher Plants, ed. M., Cresti et al. , pp. 119–24. New York: Springer-Verlag.CrossRefGoogle Scholar
Kranz, E., Bautor, J. & Lorz, H. (1991). In vitro fertilization of single, isolated gametes of maize by electrofusion. Sex. Plant Reprod. 4, 1216.Google Scholar
Mo, Y.-S. & Yang, H.-Y. (1992). Isolation and fusion of sperm cells in several bicellular pollen species. Acta Bot. Sin. 34, 688–97.Google Scholar
Read, S.M., Clarke, A.E. & Bacic, A. (1993). Requirements for division of the generative nucleus in cultured pollen tubes of Nicotiana. Protoplasma 174, 101–15.Google Scholar
Russell, S.D. (1991). Isolation and characterzation of sperm cells in flowering plants Annu. Rev. Plant Physiol. Plant Mol. Biol. 43, 189204.Google Scholar
Shivanna, K.R., Xu, H., Taylor, P. & Knox, R.B. (1988). Isolation of sperms from the pollen tubes of flowering plants during fertilization. Plant Physiol. 87, 647–50.CrossRefGoogle ScholarPubMed
Southworth, D. & Knox, R.B. (1988). Methods for isolation of sperm cell from pollen. In Plant Sperm Cells as Tools for Biotechnology, ed. H.J., Wilms & C.J., Keijzer, pp. 8795. Wageningen: PUDOC.Google Scholar
Yu, H.-S. & Russell, S.D. (1994). Male reproductive cell development in Nicotiana tabacum: male germ unit associations and quantitative cytology during sperm maturation. Sex. Plant Reprod. 7, 324–32.CrossRefGoogle Scholar