Introduction and objective. Since the role of vitamin D is essential in numerous biological processes its deficiency was suggested to be a risk factor for e.g. osteoporosis, musculoskeletal pain and spine pain. The purpose of the study was to analyse whether serum vitamin D concentration is related to pain involving the motor system in Polish postmenopausal women working in agriculture. Material and methods. The study group consisted of 1,751 post-menopausal women, aged 45–65, at least 12 months from the last menstrual period, living in rural areas and working in agriculture. The research method was self-assessment of pain involving the motor system using VAS, laboratory test of serum vitamin D concentration and a medical interview. Statistical methods included generalized linear models, analysis of variance, t test for two means in two independents, χ2 test of stochastic independence. Results. Postmenopausal women working in agriculture and suffering from pain in at least one part of the motor system were younger and lower educated, they also had higher abdominal obesity and lower serum vitamin D, compared to those without pain in any part of the motor system. Decreased serum vitamin D concentration in postmenopausal women working in agriculture is important from the aspect of a higher prevalence of pain in the thoracic spine and more severe pain in the neck spine, but not for severity of pain in the lumbar spine; higher occurrence of pain in both hands or wrists; higher prevalence and more severe pain in at least one knee; and no prevalence or severity of pain in the shoulders and elbows. Conclusion. Serum vitamin D concentration is important for the prevalence and severity of pain in the neck and thoracic spine, knees and hands or wrists, but not for the lumbar spine, shoulders and elbows.