Network interworking is a key technological bottleneck for the acquisition of mobile video. To solve this problem, based on the studies of video segmentation, compression, annotation and retrieval, the network interworking gateway for mobile video acquisition was designed which can eventually achieve the acquisition of mobile video of vegetables harmed by pests. Firstly, the 3G-324M and H.323 network protocol of video telephone network and IP network were analyzed in detail. Then, an interworking gateway of 3G-324M and H.323 was designed, which included signal gateway and media gateway. Signal gateway is responsible for handling the signal and realizing the control and communication of call level and system level between video telephone network and IP network. Media gateway is responsible for data transmission, realizing the control and communication of media level and data level between video telephone network and IP network. Finally, based on the previous research foundation and the network interworking gateway designed in this paper, a mobile acquisition model was built, and the mobile video acquisition of vegetables harmed by pests was realized. The comparative experiment was carried out on the video acquisition model. Comparing with the traditional methods, it proved that the proposed network interworking gateway and video acquisition model processed theoretical and practical value.