Agrobacterium tumefaciens is the agent that causes crown gall tumor disease on more than 140 species of dicotyledonous plants. Chemotaxis of A. tumefaciens toward the wound sites of the host plant is the first step to recognize the host. CheW is a coupling protein that bridges the histidine kinase CheA and the chemoreceptors to form the chemotaxis core signaling complex and plays a crucial role in the assembly and function of the large chemosensory array. Unlike all previously reported chemotaxis systems， A. tumefaciens has only one major che operon but two cheW homologs (atu2075 as cheW and atu2617 as cheW) on unlinked loci. The in-frame deletion of either cheW gene significantly affects A. tumefaciens chemotaxis but does not abolish the chemotaxis， unless both cheW genes were deleted. The effect of cheW deletion on the chemotaxis is more severe than that of cheW deletion. Either CheW can interact with CheA and couple it to the cell poles. The promoter activity of cheW is always higher than that of cheW under all of the tested conditions. When two cheW genes were adjusted to the same expression level by using the identical promoter， the difference between the effects of two CheW proteins on the chemotaxis still existed. Therefore， we envision that both the different molecular ratio of two CheW proteins in cell and the different affinities of two CheW proteins with CheA and chemoreceptors result in the efficiency difference of two CheW proteins in functioning in the large chemosensory array.