Background: Neofusicoccum parvum belongs to the Botryosphaeriaceae family, which groups endophytic and latent pathogens of woody plants responsible for diseases such as cankers, dieback and blight. It is a widespread pathogen with a broad host range, including agricultural, horticultural and forestry plants; therefore, it is relevant to characterize the molecular mechanisms involved in the disease caused by this pathogen. This work reports for the first time N. parvum as a pathogen of Liquidambar styraciflua. We established an in vitro pathosystem using foliar tissue to characterize the infection process through scanning electron microscopy (SEM). Because cysteine-rich proteins (CysRPs) have been studied for their important functions in plant-pathogen interactions, new CysRPs were identified for these organisms, and mRNA expression of these proteins was analyzed at early time points during the interaction.Results: After the first 24 hours post infection, the pathogen caused visible symptoms, and microscopic analysis at 16 days post infection revealed the presence of N. parvum pycnidia embedded in L. styraciflua leaf tissue. For both organisms, two databases with transcriptomic and genomic information were analyzed, and five new CysRPs were identified for each organism. The length varied between 95 and 204 amino acids, and in silico analysis revealed that all the proteins are potentially secreted. The search for conserved domains and phylogenetic analyses revealed that all the proteins are novel, including two of N. parvum that present the well-known CFEM domain. RT-qPCR analysis was conducted at 24 and 72 hours post infection, and the results showed changes in the levels of CysRP mRNAs for both the plant and the fungus at early stages during the interaction.Conclusions: N. parvum was identified for the first time as a pathogen of L. styraciflua, and this work presents an approach to comprehensively understand the molecular mechanisms involved in this interaction, highlighting the potential involvement of CysRPs of both organisms under this biotic stress.