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Seed potato farming projects are essential for increasing agricultural productivity, strengthening food security, and supporting rural livelihoods in Rwanda. However, their performance is often affected by internal organizational culture challenges such as weak communication, limited accountability, unresolved conflicts, and low employee motivation. This study examined the influence of key organizational culture aspects—communication, accountability, conflict resolution, and employee motivation—on the performance of seed potato farming projects in Rwanda. The study was guided by Organizational Culture Theory and Systems Theory and employed both descriptive and correlational research designs. The target population consisted of 420 individuals, including project managers, agronomy staff, field operations personnel, administrative staff, and smallholder farmers involved in the RAB potato seed multiplication project. A sample of 230 respondents was selected using appropriate sampling techniques. Data were collected through questionnaires, interviews, and document review. A pilot test involving 23 respondents was conducted to assess the consistency of the research instruments. The obtained results show a Cronbach’s Alpha value of 0.78, confirming reliability, and a CVI of 0.82, which exceeds the acceptable threshold for validity, meaning the research instruments were both dependable and appropriate for accurately measuring the study variables. Only items with a Content Validity Index (CVI) of at least 0.70 were retained. Data were analyzed using descriptive statistics, correlation, and regression analysis with SPSS version 28 at a significance level of 0.05.