Winter wheat （Triticum aestivum） cultivars Dongnongdongmai 1 with strong cold/freezing tolerance and Jimai 22 with weak cold/freezing tolerance were used for investigating the difference of microstructure and ultrastructure between leaves of two cultivars under low temperature stress （5℃ and -15 ℃） using optical and electron microscope. The results showed that there was no obvious difference between leaves of Dongnongdongmai 1 and Jimai 22 in microstructure. However, the difference between those leaves was distinct in ultrastructure. The grana lamella and stroma lamella were stacked regularly and arranged parallelly along the long axis of chloroplasts in cv. Dongnongdongmai 1, while the arrangement directions of thylakoids in Jimai 22＇s leaves were so irregular as to form various angles with the long axis of chloroplasts. At -15℃, the mitochondrias were swelled to be round and the structure of cristaes became blurry in both cultivars＇ leaves, while some cristaes of Jimai 22 disappeared. These results would provide theoretical evidence for selecting cold/freezing tolerant winter wheat germplasm resources