Chilling is common in nature and can damage most plant species, particularly young leaves and buds. Mulberry (Morus spp.) is an economically important food source for the domesticated silkworm (Bombyx mori). However, weather and climatic extremes, such as “late spring coldness”, seriously damage mulberry buds and young leaves. The molecular mechanism involved in the differing mulberry chilling tolerance is unclear. In the present study, we found that mSOD1, mFADII, and mKCS1 interacted with mAKR2A and that the expression of mAKR2A, mSOD, mFAD, and mKCS1 in the chilling-tolerant mulberry variety was higher than that in the chilling-sensitive variety. Unsaturated fatty acids content and superoxide dismutase (SOD) activity in the chilling-tolerant variety was higher than that in the chilling-sensitive variety. After chilling treatment, mSOD1, mKCS1 and mAKR2A expression in the chilling-tolerant variety was reduced to lower than that in the chilling-sensitive variety, whereas mFADII expression increased in the chilling-tolerant variety compared with that in the chilling-sensitive variety, suggesting that the increased expression of the molecular chaperon mAKR2A helped to maintain or prompted the chilling-related proteins in the chilling-tolerant variety.