Identifying and studying the environmental and agronomic factors are of major importance in increasing the quantity and quality of medicinal plants. Hence, this study was conducted to evaluate the effectiveness of mulch and biofertilizer application on yield and yield components, oil quantity and quality of purslane in different cropping systems. Two experiments were carried out with factorial arrangement based on randomized complete block design with three replications in 2015 and 2016. The first factor was cropping systems including Portulaca oleracea L. and Lallemantia iberica Fischer & C.A. Meyer sole cropping, the additive intercropping of purslane/dragon’s head (P100/D20, D40 and D60 %); the second factor was mulch application including with (+M) or without (-M) wheat straw mulch; the third factor was two types of fertilization including application of 100% chemical fertilizer (F1) and 50% chemical + biological fertilizers (F2). The mulching significantly affected yield, yield components, oil content and omega-3 fatty acid of purslane. The highest value of omega-3 fatty acid was observed in mulched plants. Sole cropping of purslane increased number of capsule and seed yield. The highest land equivalent ratio (1.56) was observed in D40+P100 intercropped with mulch application and 50% chemical + biological fertilizers in 2016. Therefore, the organic intercropped system with mulching is applicable in the medicinal plant production such as purslane in order to reach sustainable agricultural goals.