Abstract: In order to study the optimum mature characteristic of tobacco upper leaves for plucking, the appearance and sensory qualities of YunYan87 upper leaves were analyzed. The results showed that the date of 125 days after transplanting (M2) was the best stage of one-time plucking of upper leaves. The appearance and sensory qualities scores of the top3-1 leaves at the date of 125 days after transplanting (TM2) were 44.67 and 71.79 respectively, and the top6-4 leaves at the date of 125 days after transplanting (FM2) were 43.36 and 71.75, which were all better than those of other stages. The cytology observation of tobacco leaves indicated that the palisade tissue and spongy tissue of tobacco leaves became more loosen from the date of 125 days after transplanting (M2), with the intercellular space enlarged, organelle degraded, and the starch particles were full. The thickness and cell density of palisade tissue and spongy tissue of the top2 leaves at the date of 125 days after transplanting was 145.3 μm, 144.7 μm, 43 mm and 43 mm; The thickness and cell density of palisade tissue and spongy tissue of the top5 leaves at the date of 125 days after transplanting was 139.7 μm, 148 μm, 42/mm and 41/mm, respectively. This research might provide the theoretical base for further verifying that the M2 stage was the optimum mature characteristic of upper tobacco leaves for plucking.