Tartary buckwheat seed and especially its sprouts are rich in D-chiro-inositol (DCI). The research was to evaluate when DCI was most accumulated in tartary buckwheat sprouts. In addition, we explored the activity and expression pattern of α-galactosidase during tartary buckwheat seed germination. The results showed that DCI contents steadily increased at early stage of germination and reached the highest level of 33.42 µg/seed at 24 h during the 72 h trail. However, the total fagopyritol contents sharply decreased from 214.6 µg/seed to 46 µg/seed at the end of the germination. The activity of acid α-galactosidase increased gradually to the peak of 0.36 nkat/seed at 24 h after the primed seed imbibition. We cloned the gene fragment of α-galactosidase in tartary buckwheat for the first time. The deduced amino acid sequence is 93% identical to that of P. vulgaris. The quantitative PCR result of gene expression pattern was consistent with its enzyme activity during seed germination.