Abstract: In order to provide suitable soybean accessions in southern Huanghuai, 107 soybean quality, yield data and yield related traits data from the Henan Province, Anhui Province and Jiangsu Province was collected in 2011 and 2012, and the key yield limiting factors were determined with correlation, stepwise regression and path analysis. The results indicated that, the average yield of soybean lines was 2601.73 kg/hm2 in 5 test loci. Compared to the control ‘Zhonghuang13’, 48 soybean lines showed increase yield, accounting for 44.86%; the average protein content was higher and the high protein and fat content (total protein plus fat ≥ 63% and fat ≥ 20%) accessions was 70% or more in all 5 test loci. The correlation analysis showed the yield had extremely significantly negative relation with protein, fat and protein plus fat content (r=-0.27, r=-0.23, r=-0.48), had extremely significantly positive relation with effective branch number per plant, pod number per plant, seed number per plant and seed weight of per plant respectively. Partial correlation analysis also showed that, soybean yield and seed number per plant had extremely significantly positive relation. According to the standard regression coefficient of yield, seed number per plant was the primary determinant factor that effected soybean yield in southern Huanghuai. Path-coefficient analysis revealed that, the direct effect of seed number per plant was the largest. The protein and fat content had significantly positive relation with plant height and 100 seed weight (r=0.20, r=0.23), had significantly negative relation with effective branch number per plant (r=-0.19), had extremely significantly negative relation with pod number per plant, seed number per plant and seed weight of per plant respectively (r=-0.29, r=-0.44, r=-0.37). So, the breeding should pay attention to the balance between the various characters, we should think about the various agronomic characters of soybean exists the mutual influence at each other at the same time.