METHOD FOR TRANSFORMATION OF CELL OF GRAMINEOUS PLANT USING SPHEROPLAST OF ESCHERICHIA COLI

摘要

PURPOSE:To carry out transformation of gramineous plant readily, by using a spheroplast of Escherichia coli having a desired gene with a protoplast of a cell derived from gramineous plant and transducing the gene to the cell of gramineous plant. CONSTITUTION:A cell is treated with lysozyme by the use of Escherichia coli in logarithmic growth phase cultivated in a medium such as bouillon medium, glucose M9 medium, etc., capable of multiplying Escherichia coli to give a spheroplast of Escherichia coli having a desired gene. On the other hand, leaves, roots, buds or culture cell of gramineous plant are treated with an enzyme solution obtained by dissolving cellulase and pectoliase in mannitol (0.4mol) to give a protoplast of gramineous plant. The spheroplast of Escherichia coli is fused with the protoplast of gramineous plant and the gene contained in Escherichia coli is transduced to a cell of gramineous plant to carry out transformation.