Plant & Animal Genome IX Conference

Poster: Sequencing & EST
P01_76.html

GENE DISCOVERY IN COTTON FIBER INITIATION USING EST AND MICROARRAY APPROACHES

CSIRO Plant Industry, GPO Box 1600, ACT2601, Australia

Cotton fibers are single elongated cells that develop from epidermal cells of ovules. Despite the economic importance of cotton fiber, little is known about the molecular basis of fiber initiation. Random cDNA sequencing and microarray global gene expression approaches are being undertaken at Plant Industry to uncover genes involved in fiber initiation. Two cDNA libraries (CHX and OCF) derived from cotton ovules at different developmental stages were constructed. The CHX cDNA library was developed from pooled ovules of -3 DPA to 0 DPA (Days Post Anthesis). These ovules were treated with cycloheximide (a protein synthesis inhibitor). This treatment is believed to stablise some labile mRNAs when applied prior to RNA isolation and cDNA synthesis. The OCF library was from 0 DPA ovules and has also been normalised. About 3000 cDNA clones from CHX, OCF and normalised OCF libraries have been sequenced. Preliminary sequence data analysis showed that about 28%, 24% and 30% clones from CHX, OCF and normalised OCF libraries respectively were novel genes or genes without known functions. Amplification of the cDNA clones by PCR is currently in progress and about 10,000 cDNA clones will be arrayed on glass slides to be used in gene expression studies.