Two small RNA libraries were generated from micropropagated ‘Muscat Hamburg’ (Vitis vinifera) plantlets under normal and low temperatures (4 °C). A total of 163 known miRNAs and 299 putative novel miRNAs were detected from two small RNA libraries by Solexa sequencing. Forty-four cold-inducible miRNAs were identified through differentially expressed miRNAs (DEMs) analysis; among which, 13 belonged to upregulated DEMs while 31 belonged downregulated DEMs. This study indicated that a diverse set of miRNAs in V. vinifera are cold-inducible and may play an important role in cold stress response. Examination of small RNA populations in grapevine under cold treatment and none cold treatment.