Torenia Fourinei. Transformation With Fusion Gene Of Gfp:mtn And Expression Of Chimeric Protein In Torenia Fourinei.

The actin cytoskeleton, a dynamic network of intracellular proteinaceous structural elements participates in many physiological activities in plant cell. The actin cytoskeleton polymerization and depolymerization is regulated by a number of actin-binnding protein rapidly and spatio-temporally.Talin is a important F-actin binding protein , Two functions of intact talin are the control of actin filament length and the cross-linking of actin polymers. The binding of F-actin to talin significantly increases actin filament stiffness. In addtion to, Talin nucleates actin assembly and binds to vinculins and a host of actin binding proteins and other proteins mainly at the focal adhesion.As a report probe, the green fluorescent protein(GFP) had been broadly employed in research works. The fusion gene ofgfp:mtn (mTn is binding domain of microfilament binding protein talin, which can show the microfilament in living cell) was transferred to Arabidopsis thaliana. Stable expression of GFP-mTn can visualize the actin cytoskeleton in different types in living cells without affecting cell morphology and function.Toreniafournieri. has a naked embryo sac that protrudes from the micropyle into the cavity between the funiculus and placenta. As embryo sac is mature, the egg cell, the two synergids and about half the central cell are located outside the micropyle. Therefore, Toreniafournieri is a very suitable model plant for studies of double fertilization in living material. We transferred the fusion gene ofgfp:mtn to Toreniafournieri by leaf disc transformation mediated by Agrobacterium and gain transgenic plant. Using Confocal Laser Scanning Microscopy (CLSM),we observed the distribution of chimeric protein in different kinds of tissues and cells in transgenic Toreniafournieri. We find the chimeric protein in leaf epidermal cells, stomatal guard cell and root epidermal cell. Actin filaments can be visualized clearly in guard cell, in guard cells of open stomata under light, actin filaments arrange reticulate randomly in cytoplasm. In guard cells of closed stomata under darkness, actin filaments arrange curly along the longitude of guard cell. Green fluorescence can be visualized in leaf epidermal cells and root epidermal cell in few of transgenic Toreniafournieri. Actin filaments can not be visualized in other cell types, including stem epidermal cell, root fair cell and pollen.We transferred the hetero-logous gene into Toreniafournieri successfully. This method laid a foundation for study of double fertilization of angiosperm by transgenic tecnology. The transgenic Toreniafournieri we got suitable provides a material to studies dynamics of actin filament in stomatal guard cell.