An embodiment of the invention discloses a mask plate, a columnar spacer manufacture method and a substrate manufacture method, and relates to the technical field of display. By means of the invention, the segment difference between each main columnar spacer and each auxiliary columnar spacer and the size of the auxiliary columnar spacers can be considered simultaneously. the mask plate comprises first light-transmitting zones, second light-transmitting zones, third light-transmitting zones and fourth light-transmitting zones, wherein the width of each of the first light-transmitting zones and the third light-transmitting zones is d1; the width of each of the second light-transmitting zones and the fourth light-transmitting zones is d2; one second light-transmitting zone, one third light-transmitting zone and one fourth light-transmitting zone are arranged sequentially between two adjacent first light-transmitting zones; the width of a shading zone between each second light-transmitting zone and each first light-transmitting zone as well as between each fourth light-transmitting zone and each first light-transmitting zone is d3; the width of a shading zone between each third light-transmitting zone and each second light-transmitting zone as well as between each fourth light-transmitting zone and each third light-transmitting zone is d4; and d4 is equal to a quotient by dividing 1 by a product after multiplying 2 and the sum of d1, d2 and 4*d3.