A display substrate, a driving method thereof and a display device are disclosed. The display substrate includes repeatedly arranged pixel groups. Each pixel group includes a first sub-pixel group, a second sub-pixel group, a third sub-pixel group and a fourth sub-pixel group. Each of the first sub-pixel group, the second sub-pixel group, the third sub-pixel group and the fourth sub-pixel group includes four pixel columns. The display substrate, the driving method thereof and the display device reduce the number of sub-pixels in the whole display device and hence reduce the difficulty in the manufacturing process of the display device and the cost on the premise of ensuring that the display device obtains a high resolution.