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Isolation of two populations of sperm cells and microelectrophoresis of pairs of sperm cells from pollen tubes of tobacco (Nicotiana tabacum)

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Abstract

Prior research has indicated that the two sperm cells of Nicotiana tabacum are dimorphic, suggesting that they may participate in preferential fertilization during in vivo fusion with the egg and central cells. To probe the mechanism of potential preferential fertilization in this plant, it will be necessary to use modern sensitive molecular techniques. For this purpose, two individual populations of two sperm cells, constituting the Svn (associated with the vegetative nucleus) and Sua (unassociated with the vegetative nucleus), were isolated in the thousands from tobacco pollen tubes with a micromanipulator as a preliminary step toward research on gametic recognition using molecular techniques. Microelectrophoresis of paired sperm cells from a single pollen tube was conducted at different developmental stages. Sperm cells isolated from 1-, 2-, 3- and 4-cm stylar lengths migrated to the negative pole, with the Sua displaying significantly greater electrophoretic mobility than the Svn, reflecting a more positively charged cell surface on the Sua. The sperm cells isolated from 1-cm style are very sensitive to electron potential in an electrophoretic field, presumably reflecting that they are still in a young state. Differences in cell surface charge between the Sua and Svn may be related with cell fate during fertilization.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China.

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Authors and Affiliations

  1. Key Laboratory of Cell Biology and Tumor Cell Engineering of Ministry of Education, School of Life Sciences, Xiamen University, 361005, People’s republic of China

    YH Yang, YL Qiu, CT Xie & HQ Tian

  2. Department of Botany and Microbiology, University of Oklahoma, Norman, OK, 73019, USA

    Z Zhang & SD Russell

Authors
  1. YH Yang
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  2. YL Qiu
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  3. CT Xie
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  4. HQ Tian
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  5. Z Zhang
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  6. SD Russell
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Corresponding author

Correspondence to HQ Tian.

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Supported by National Natural Science Foundation of CHINA (30170060)

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Yang, Y., Qiu, Y., Xie, C. et al. Isolation of two populations of sperm cells and microelectrophoresis of pairs of sperm cells from pollen tubes of tobacco (Nicotiana tabacum). Sex Plant Reprod 18, 47–53 (2005). https://doi.org/10.1007/s00497-005-0248-x

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  • DOI: https://doi.org/10.1007/s00497-005-0248-x

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