Purpose: Teachers are important stakeholders in supporting children’s physical literacy (PL), yet teachers’ perception of PL assessment is underexplored. Method: Utilizing a mixed-methods design, 122 primary school teachers (of children aged 5–12 years) in Australia completed an online survey, followed by nine interviews. Results: Teachers who favored assessment (58%) tended to report assessing PL in children (χ²[1, N = 110] = 7.025, p = .008). Those who reported assessing PL (also 58%) were more confident to do so (χ²[2, N = 109] = 10.540, p = .005). Teachers considered movement skills, engagement and enjoyment, relationships, and safety and risk as the most important elements for assessing PL. Qualitative data showed nonsupport for PL assessment stemmed from skepticism regarding relevance of assessment, appropriateness of assessment, and views that the curriculum and PL framework were implicitly linked. Conclusion: Professional development, resources, and suitable PL teacher assessments can upskill teachers’ knowledge, confidence, and reduce barriers in implementing PL assessments.