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Gou, Xiaoping [1], Yuan, Tong [1], Wei, Xiaoping [1], Russell, Scott D. [1].

Differential gene expression between the sperm cells of a flowering plant, Plumbago zeylanica.

Mature pollen of Plumbago zeylanica contains two sperm cells (Sua and Svn) and one vegetative cell. The two sperm cells have different organelle complements and fuse with different female target cells. The Sua contains more plastids and fuses with the egg cell to form the zygote. The Svn contains more mitochondria and fuses with the central cell to produce the endosperm. To discriminate gene expression differences between the two sperm cells, we constructed subtractive cDNA libraries of these two sperm cell types for microarray screening. The inserts were PCR amplified and spotted onto glass slides for microarray screening. 2304 clones of each subtractive cDNA library were examined using subtracted and unsubtracted cDNA probes. Subtractive hybridization made it easier to identify those genes differentially expressed in these two sperm cell types. Several hundred clones that have differential expression patterns in the two sperm cell types have been identified according to microarray data. Expression patterns of selected clones have been confirmed by RT-PCR and realtime RT-PCR screening. From these candidates, we have obtained several one-sperm-cell-specifically expressed genes, and their expression specificity has been confirmed by whole mount in situ hybridization in pollen. One sequence is homologous with cytokinin synthase in Arabidopsis and is specifically expressed in Svn. The abundance of cytokinin synthase in the Svn sperm, which fuses with the central cell to form the endosperm, may differentially deliver paternal transcripts contributing to high levels of cytokinin production suspected to stimulate early endosperm activation.

Related Links:
Russell Lab Page
Plumbago zeylanica cDNA sequencing project

1 - University of Oklahoma, Department of Botany and Microbiology, 770 Van Vleet Oval, Norman, Oklahoma, 73019, USA

EST sequencing
sperm cell gene expression
sperm dimorphism
Plumbago zeylanica
double fertilization.

Presentation Type: Poster
Session: 32-155
Location: Special Event Center (Cliff Lodge)
Date: Tuesday, August 3rd, 2004
Time: 12:30 PM
Abstract ID:1135

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