The Auxin And Auxin Binding Protein1Regulation Of Vesicular Transport In The Tobacco BY2Cell

The Auxin Binding Protein1plays an important regulation role in cells elongation. It is also one of the important pathways of auxin singal transduction. Accumulated researchs showed that plants cells quick response to auxin signal lead to cells elongation were not involed in gene expression process. Plant cells elongation inevitably involved a specific process in cell wall synthesis, the transportation of quantities memberane components, and the cytoskeleton assembly. Using the plant model cell lines of tabacco BY2as materiers, we labeled SCAMP2(Secretory Carrier Membrane Protein2) fusion with green fluorescent protein (GFP) as vesicle molecular markers. We studied the regulation of cells vesicles transport process which involved in ABP1responses to auxin.SCAMP2can be used as the suitable markers of the vesicular transport research for its widely distributed in cytoplasmic membrane and intracellular membrane system and with the characteristics of constitutive. In order to study function and effect of ABP1at the process of vesicular transport we constructed an overexpression and antisense inhibition expression of ABP1recombinant. The chaimera NtSCAMP2-GFP was first transformed into BY2cells. It is observed that the membrane vesicles had been labeled effectively in the transformed cells. The NtABP1cDNA was cloned into an induciable Ti vector pER16in sence or antisence direction. The recombinants were transformed into the NtSCAMP2-GFP labeled BY2cell line via Agrobacterium tumefaciens LBA4404co-culture transformation. The sense or antisense ABP1in pER16can be expressed in the induction of the sterol hormones. We used Dexamethasone(DEX) as inducer of the transgenic cells, combination with the auxin treatments, the cells are observed under fluorescence microscopy or scan laser confocal microscope. The result showed that the fluorescence signal in the nucleus and near the cell matrix was significantly enhanced but with little change in the cytoplasmic membrane and the two cell intervals when NtABPl was overexpression. The location near the nucleus had a strong fluorescence signal, while the cytoplasmic membrane and two cells intervals of the fluorescence signal was significantly reduced when ABP1was antisense inhibited. This demonstrated that expression quantity alteration of ABP1in cells can impact on the vesicular transport directly. The impact was not only involved in the inhibition of endocytic vesicle assembly of clathrin but also involved in the efflux transport of vesicles. These results can provide new evidence for vesicles transport which regulated by ABP1.