It exists gender role among husbands and wives in African rural societies, they perform their own duties and try to secure their livelihoods. In recent years, the number of female-headed households tends to increase globally. Female-headed households are low socio-economic status than male-headed households. In Republic of Uganda of research country, female-headed households accounted for 30% of all households. This paper aims to clarify the economic differentiation among female-headed households focusing on the causes of separation from their spouses, the landownership and income and expenditure in households. Through field survey, it was clear to exist the gap of economic conditions among female-headed households, and the landownership was one factor to give raise to economic differentiation. The economic resources of the spouses and the separation from the spouses greatly influenced whether women can obtain the landownership or not. Landowing households engage in agricultural work and cash income-generating activities, and secure their own livelihoods. On the other hand, other women are unable to own land, and instead try to rent land. Even though women engage in agricultural work in rented land, it is difficult for them to cover the living costs. It suggests that the economic differentiations are expanding among female-headed households in this region. It must consider how to support vulnerable women putting into the lowest socio-economic position in society.