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

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.

[1]  T. Kuroiwa,et al.  Technical advance: single pollen typing combined with laser-mediated manipulation. , 1999, The Plant journal : for cell and molecular biology.

[2]  L. Liotta,et al.  Immuno-LCM: laser capture microdissection of immunostained frozen sections for mRNA analysis. , 1999, The American journal of pathology.

[3]  M. Jackson,et al.  Gene expression profiles of laser-captured adjacent neuronal subtypes , 1999, Nature Medicine.

[4]  Petra von Wiegen,et al.  Endosperm Development after Fusion of Isolated, Single Maize Sperm and Central Cells in Vitro , 1998, Plant Cell.

[5]  T. Sasaki,et al.  The rice genome project in Japan. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[6]  K. Schütze,et al.  Microbeam MOMeNT: non-contact laser microdissection of membrane-mounted native tissue. , 1997, The American journal of pathology.

[7]  B. Stone,et al.  Cell wall (1 → 3)- and (1 → 3, 1 → 4)-β-glucans during early grain development in rice (Oryza sativa L.) , 1997, Planta.

[8]  L. Liotta,et al.  Laser Capture Microdissection , 1996, Science.

[9]  T. Dresselhaus,et al.  Representative cDNA libraries from few plant cells. , 1994, The Plant journal : for cell and molecular biology.

[10]  K. Hoshikawa Anthesis, fertilization and development of caryopsis , 1993 .

[11]  Kunisuke Tanaka,et al.  The Site of Synthesis and Accumulation of Rice Storage Proteins , 1986 .

[12]  M. Okamoto,et al.  Enzymic mechanism of starch breakdown in germinating rice seeds : 11. Ultrastructural changes in scutellar epithelium. , 1982, Plant physiology.

[13]  Kunisuke Tanaka,et al.  Isolation and Characterization of Two Types of Protein Bodies in the Rice Endosperm , 1980 .

[14]  T. Akazawa,et al.  Enzymic mechanisms of starch breakdown in germinating rice seeds: 7. Amylase formation in the epithelium. , 1979, Plant physiology.