Artificial Intelligence (AI) has made its presence felt in various areas of human activity, one of which is education. However, the extent to which AI is used is not the same, as its integration is more widespread in higher, secondary and primary education, while in pre-school education AI is used to a lesser extent. The integration of digital technologies, and therefore AI, from pre-school level is considered important because it has long-term positive effects and addresses stereotypes early on. In any effort to implement innovations in education, a key success factor is the appropriate preparation and support of teachers. This article will present the results of the investigation of the training needs of kindergarten teachers in Greece regarding the use of AI applications in their teaching interventions. For the needs of the investigation, a survey of a sample of 85 kindergarten teachers, who applied for participation in professional development program for AI, in Greece was conducted. More specifically, participants were asked to answer a questionnaire consisting of seven different latent variables to be explored. These variables are as follows: Perceived Usefulness (PU), Attitude towards using AI (AT), Anxiety about AI, Confidence in teaching AI (CON), Readiness of AI (RE), Relevance of AI (RA), and Behavioral Intention (BI). The data analysis provides insights into kindergarten teachers’ educational needs regarding the use of AI applications in their teaching practice. Teachers’ responses are being processed and the initial findings seem interesting for designing training activities to prepare kindergarten teachers.