The accurate site for vitellogenin synthesis was investigated in two representative shrimp species Litopenaeus vannamei and Macrobrachium rosenbergii with real-time PCR in this experiment. Based on histological observation, the ovarian development was divided into six stages in L. vannamei (oogonium proliferation, previtellogenic, primary vitellogenic, secondary vitellogenic, mature, and recovery stages) and five stages in M. rosenbergii (previtellogenic, primary vitellogenic, secondary vitellogenic, mature, and gravid stages). It was found that both L. vannamei and M. rosenbergii expressed Vg-mRNA in hepatopancreas and ovary. In L. vannamei, the relative quantity of Vg-mRNA expression of each developmental stage was 1.1, 5.9, 10.4, 26.9, 85.3, and 1.5 in ovary, and 1.3, 3.2, 7.1, 37.2, 51.6, and 1.0 respectively in hepatopancreas. In M. rosenbergii, the relative quantity of Vg-mRNA expression of hepatopancreas in each ovarian developmental stage was 3.4, 12.6, 15.2, 38.9, and 2.9 compared to ovary 1.0, 1.3, 1.7, 4.8, and 1.5, respectively. In all examined shrimps the Vg-mRNA expression decreased sharply to the minimum at the last development stage.