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Expressed sequence-tag analysis of tobacco sperm cells reveals a unique transcriptional profile and selective persistence of paternal transcripts after fertilization

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Abstract

Transcript analysis of male gametes of Nicotiana tabacum was conducted to gather gene expression data regarding the specialization of male germ cells and transmission of paternal transcripts during fertilization. We constructed a tobacco sperm cell cDNA library yielding 1,864 expressed sequence tags representing 1,050 clusters; 37.2% of these clusters have no homologs in GenBank, and 42% did not match any functionally classified protein. A comparative analysis of tobacco sperm transcripts with those of Arabidopsis and maize confirms that some genes are conserved in sperm specialization, while some are distinct to tobacco germline cells. Using reverse transcription–PCR (RT–PCR) of selected transcripts, we evaluated expression of sperm-obtained sequences in vegetative tissue, isolated egg cells, zygotes, and two-celled proembryos, identifying sperm cell–specific transcripts as potential markers for fertilization analysis. We further confirmed that two clusters of sperm transcripts were detected in zygotes about 10 h after fertilization, offering new examples of apparently paternally transmitted transcripts that may be involved in egg cell activation and/or early embryogenesis.

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Acknowledgments

The project was supported by the ‘973’ Project (2007CB108700), National Natural Science Fund of China (30821064; 30771131) and The Chinese 111 project B06018. We thank Scott D. Russell (University of Oklahoma), Thomas Dresselhaus (Universität Regensburg) for their critical reading of the manuscript and Sheila McCormick (University of California, Berkeley) for useful discussion.

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Authors and Affiliations

  1. Key Laboratory of the Ministry of Education for Plant Developmental Biology, College of Life Science, Wuhan University, 430072, Wuhan, China

    Hai-Ping Xin, Xiong-Bo Peng, Jue Ning, Ting-Ting Yan, Li-Gang Ma & Meng-Xiang Sun

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  1. Hai-Ping Xin
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  2. Xiong-Bo Peng
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  3. Jue Ning
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  4. Ting-Ting Yan
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  5. Li-Gang Ma
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Corresponding author

Correspondence to Meng-Xiang Sun.

Additional information

Communicated by Scott Russell.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Primers used for RT–PCR expressional analysis of tobacco sperm ESTs. (DOC 38 kb)

497_2010_151_MOESM2_ESM.pdf

BLAST matches for clusters in Nicotiana tabacum sperm ESTs when compared to sperm microarray of Arabidopsis, maize sperm ESTs and Plumbago sperm ESTs. (PDF 155 kb)

Clusters that have a significant match with sperm cell transcripts in both Arabidopsis and maize. (DOC 83 kb)

497_2010_151_MOESM4_ESM.tif

RT–PCR analysis of the presence of vegetative cell specific Nt59 transcripts in isolated sperm cells. During PCR, 25 cycles were used for GAPDG and 30 cycles for Nt59. (TIFF 521 kb)

497_2010_151_MOESM5_ESM.tif

Expression analysis for sperm transcripts in anther and pollens. cDNA was synthesized from isolated sperm cells, matured anthers and pollens of tobacco, respectively. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) was used as a control. During PCR, 25 cycles were used for GAPDH and 35 cycles were used for transcripts derived from sperm cell cDNA library. (TIFF 2046 kb)

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Xin, HP., Peng, XB., Ning, J. et al. Expressed sequence-tag analysis of tobacco sperm cells reveals a unique transcriptional profile and selective persistence of paternal transcripts after fertilization. Sex Plant Reprod 24, 37–46 (2011). https://doi.org/10.1007/s00497-010-0151-y

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  • DOI: https://doi.org/10.1007/s00497-010-0151-y

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