Author
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Bausher, Michael |
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Niedz, Randall |
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Submitted to: Meeting Abstract
Publication Type: Abstract Only Publication Acceptance Date: 3/9/2004 Publication Date: 7/1/2004 Citation: Bausher, M.G., Niedz, R.P. 2004. Gene expression during normal embryogenic induction using semi-permeable membranes in citrus sinensis. Meeting Abstract. Interpretive Summary: Technical Abstract: In an earlier work, we describe the use of semi-permeable membranes to induce normal somatic embryogenic tissue to form viable somatic embryos (InVitro Cell Dev-Plant, 38:552-557, 2002). To further characterize this system, we examined the gene expression profiles of the induced and non-induced tissues using nylon filter arrays constructed from cDNAs of a Citrus sinensis L. Osbeck expression library (array of 3456 cDNAs). In eight biologically replicated experiments, we were able to account for the systematic and biological variation between the replicates and consequently determined several significant differences in gene transcription levels during membrane-induced embryogenesis. Two cDNAs, demonstrated to be induced >1.5 fold during somatic embryo formation, have no significant match to public databases suggesting the need for further characterization of these genes and their role in embryogenesis. In addition, the analysis revealed two cDNAs with suppressed transcription (<1.5 fold) during somatic embryo formation that have strong similarities to sucrose synthase and extension. The use of nylon filter cDNA arrays in combination with quantitative RT-PCR can a powerful tool to detect small differences in gene transcription when properly replicated. The ability to lessen biological and systematic variability or at least account for the variables and determine the part they play in the outcome of the results is absolutely necessary in gene profiling experiments. |
