Extended Abstract Introduction: Spinach (Spinacia Oleracea L.) belongs to Chenopodiaceae and is one of the most important leafy vegetables in the world. Spinach is a valuable human food source because its leaves are rich in minerals, vitamins, antioxidants, and phenolic compounds. Weeds are one of the significant challenges for spinach production. They can lead to a decrease in commercial biomass and spinach quality. The incidence of herbicide resistance in spinach fields has led to the use of environmentally friendly methods such as cover crops. These plants can cause physical suppression of the weeds by creating adequate biomass and developing a canopy. The aim of this study was to provide a quantitative study of the interactions of the cover crops (such as oat and daikon radish) and hand-weeding treatments on the suppression of the weeds and their effect on the vegetative growth and yield of spinach.Materials and Methods: The experiment was conducted in the crop years of 2020-2021 and 2021-2022 at the Research Farm of the Faculty of Agriculture and Natural Resources of the University of Mohaghegh Ardabili with 1350 meters above sea level and geographical coordinates 38˚ 19´ East and 48˚ 20´ North with cold and semi-arid climates. The experiment was performed as a randomized complete block design with three repetitions. Experimental treatments included the first factor of oat (Avena sativa L.), daikon radish (Raphanus sativus var. Longipinnatus), their intercropping (with 50 % seed ratio), and control treatment (without cover crops). The second factor consisted of hand-weeding and non- weeding. The seed rate of oat and daikon radish was 100 and 20 kg per ha, respectively, and were planted manually at a 3×4 meter plots. In this experiment, the seeds of Spinach Viroflay were used. Seeds were sowed manually among the residues of cover crops at 1.5 cm depth. Due to the amount of precipitation in two years, irrigation was performed if needed. The inter and intra-row spacing was 20 and 10 cm. Hand-weeding was done from the planting date to the final harvest of spinach. The investigated traits included dried biomass of cover crops, the density of dominant weeds, and Simpson, Shannon-Weaner, and Margalef indices.Results and Discussion: The biomass of cover crops was significantly affected by planting systems, seed density, and year (P ≤1 %). The results showed that the highest biomass of cover crops (538.83 gm-2) was obtained from oats. Also, the lowest biomass (325.83 g.m2) belonged to daikon radish. The dominant weeds were consisted of three species such as lamb's quarters (Chenopodium Album), italian bugloss (Anchusa Italica Retz), and common fumitory (Fumaria officinalis). The results showed that oat in two years had the lowest weed biomass (10.64 g.m2) and density of lamb's quarters, italian bugloss, and common fumitory (4.33, 5.00, and 4.25 number/m2). The maximum weed control efficiency index (57.67 %) was obtained from the oat monoculture. Also, the highest weed control efficiency index (51.47 %) was obtained from the hand-weeding. Margalef's index decreased in the first and second sampling times compared to 5.55 % and 9.85 % relative to the first year. The highest index of Shannon-Weaner's diversity belonged to control and daikon radish (0.47 and 0.47). The highest baby spinach yield and final yield in the first and second phases of sampling in two years were obtained from oats treatment (with 567.05 and 944 g.m2, respectively) and then from their intercropping (390.04 and 786.33 g.m2).Conclusion: The results showed that oat and its combination with daikon radish had the highest biomass compared to daikon monoculture. The effect of oat biomass on the decrease in density and biomass of the total weeds was significant, which was due to the amount of oat seeds consumed compared to daikon radish. In addition, hand-weeding reduced the density and biomass of the weeds more than non-weeding. Hand-weeding led to the lack of opportunity for weeds' growth. As such, the improvement of the baby spinach and the final yield was due to the hand-weeding of the weeds and the presence of the cover crops biomass in the experiment. The results of are inferred that oat monoculture is suitable both in terms of reducing physical competition of horticultural plants with weeds and improving spinach yield.