Freezing injury is one of the important abiotic factors affecting wheat yield. It is difficult to prevent and control in wheat production. In this study, we used the spring cultivar Zhongmai 8444 as the test material, carried out low temperature stress treatment in the early stage of gestational panicle to deal with them. And RNA-seq sequencing was carried out in young stems. Based on the transcriptome difference analysis data obtained, genes with upregulated expression and function related to cold resistance after low temperature stress treatment were selected as candidate genes for further functional verification. Here, we predicted that TaNAC2D (GQ231952.1), TaADF (XM044535327), TaAP2 (XM_037623978.1), TaCALB (XM_037552257.1), TaCOBRA (XM_037564732.1), TaHF (XM_044576055.1) can have an impact on the cold resistance of Zhongmai 8444. We used BSMV-VIGS technology to instantly silence the cold-resistant related genes of Zhongmai 8444, and then rapidly verify the function of the candidate genes by low temperature stress treatment. We found that Zhongmai 8444 compared with materials of silenting TaNAC2D (GQ231952.1), TaADF (XM044535327), TaAP2 (XM_037623978.1) in the environment of -5°C, the leaf cold resistance was significantly reduced and the leaf wilting was characterized. The relative conductivity of the third leaf was also determined that the silent TaNAC2 (GQ231952.1), TaADF (XM044535327), TaAP2 (XM_037623978.1) was significantly higher than that of the negative control group. The expression of TaNAC2D (GQ231952.1) was determined by qRT-PCR. The gene expression of Zhongmai 8444 leaves after silencing the TaNAC2D (GQ231952.1) was significantly lower than that of the negative control group. This study can provide a theoretical basis and lay the foundation for the genetic improvement of wheat cold resistance traits.