This paper took the mulberry leaves of Yun-sang NO.1 as experimental material, under the conditions of different cold storage temperature and different cold storage time, refrigerated the mulberry leaves of different planting patterns and different leaf position, measured the protein content in mulberry leaves, analyzed the changing rule of the protein. The results show that under the zero and hundred degrees centigrade of storage temperature conditions, each type of mulberry leaves storages eight days later, the protein content in the zero degree centigrade is significantly higher than that in the hundred degree centigrade ; the protein content of quadrilateral mulberry is significantly higher than that in the campus mulberry and dense planting mulberry, the protein content of upper leaves is significantly higher than that in the middle leaves and lower leaves; with the extension of storage time, the average protein content shows a downward trend, from fast to slow as follows： upper leaves of quadrilateral mulberry, upper leaves of campus mulberry, middle leaves of quadrilateral mulberry ,upper leaves of dense planting mulberry.