When the model was applied to discriminate the sex of silkworm pupae of new species, the accuracy was not satisfied. Rebuilding the model need abundant samples and was time consuming. Updating the original model with few samples was usually used, which could also gave high performance. Hence, different model updating strategies were explored via visible (VIS)/near-infrared (NIR) hyperspectral imaging (HSI) spectroscopy in this paper. The calibration model including 150 samples was established on two species (221B-403, 223B-404) and the prediction set had 50 samples. Different variable selection methods including uninformative variable elimination (UVE) and synergy interval-partial least-squares (SiPLS) were adopted. Then partial least squares-discriminant analysis (PLS-DA) and extreme learning machine (ELM) were used to build the model. The best performance were given by UVE-PLSDA and SiPLS-PLSDA models with the accuracy of 98%. When SiPLS-PLSDA model was explored to discriminate the sex of new species “9312-shanheB”, the accuracy was just 70.83%. Different sample selecting methods, containing Kennard Stone (KS) algorithm, Manhattan distance, Euclidean distance and Chebyshev distance, were used to update the original calibration model. The updated SiPLS-PLSDA model based on KS gave the highest performance with 100% when used to discriminate the sex of silkworm pupa with different species in the same year. In addition, the updated model reached the accuracy of 94.44% when used to discriminate the sex of silkworm pupa with different species in different year. It indicated that the proposed updating strategy could be further effectively explored to differentiate the sex of other new species.