Grape (Vitis L.) is one of the most important economical fruit trees in China. However, a shattering rachis-like phenotype in some varieties is un-desirable, particularly during long-distance transport and storage. Breeding for storage-resistant varieties is one of the most effective measures to prevent this problem. In this study, six hundreds of accessions from Zhengzhou National Grape Germplasm Repository were evaluated for the pulling force of fruit stalk with a digital tension machine. Whether pulling force of fruit stalk correlated to fruit handle, fruit pedicle and fruit brush lengths were analyzed as well. The results showed that the pulling force of six grape varieties showed a decreasing trend during fruit ripening. The pulling force of fruit stalk of hard flesh is stronger than that of soft flesh varieties. The pulling force of 600 grape accessions ranged from 0.72 to 9.16N, showing a continuous distribution consistent with positive skewness distribution. The pulling force was distributed into weak (P≤3N), medium (3N＜P≤6N) and strong (P＞6N) 3 classes, with respective proportion of 73.7%, 23.8% and 2.5%. It showed that most grape accessions tested in the germplasm resources have weak pulling forces. Several grape varieties showed strong pulling force of fruit stalk, including Red Globe, MuDanhong, Mathias Aromatic and so on. The pulling force of fruit stalk in different varieties was positively correlated with the single grain weight, fruit stalk and fruit brush lengths, unit cross-section area of fruit stalk, fruit pedicle and fruit brush lengthes. Moreover, the pulling force negatively correlated with the ratio of unit cross-section area of fruit brush versus berry weight. Taken together, this research provided valuable information on characterizing the pulling force of grape germplasm resources in future breeding for new grape varieties.