Separation of Specific Cells Using a Laser Micro-dissection System and Detection of mRNA in Developing Rice Endosperm Tissue

Abstract

Cloning of the expressing gene in the target cell is important for understanding the molecular mechanism of specific gene expression. Laser micro-dissection techniques allow the precise separation of a single target cell from morphologically distinct cells. We describe here an excellent method for separating a single cell or a cell cluster consisting of a few cells from a small target area of plant tissues. In rice endosperm tissues, the aleurone layer and starchy endosperm tissue are differentiated from a single endosperm mother cell. We succeeded in dissecting a specific cell area from endosperm and aleurone tissue (7 days after flowering). This information will be useful for studying cell differentiation and developing mechanisms of endosperm tissue. RT-PCR analysis for mRNAs for actin and glutelin confirmed the purity of the dissected cells.