The purpose was to evaluate the response of red pitaya fruit quality and yield to organic fertilizer mixed with biochar. A field experiment was conducted with five-year-old “Tainong no.2” red pitaya，three levels of chaff organic fertilization amounts and three mixture proportion of bamboo biochar were included. The pitaya fruit was harvested in six batch in 2019，and the fruit from four batch were selected as samples in this study. The effects of different fertilization treatments on the yield and quality，including reducing sugar content，soluble protein content，vitamin C(Vc) content，soluble solids content and anthocyanin content in flesh as well as edible rate of fruit，were investigated. The results showed that the organic fertilizer significantly affected the contents of soluble protein，anthocyanin，soluble solids contents and the edible rate of fruit(P<0.05). The biochar significantly affected the contents of soluble protein，anthocyanin，and soluble solids in the flesh in the first batch，and reducing sugar and Vc in the second batch(P<0.05). The coupling of organic fertilizer and biochar significantly affected the contents of reducing sugar，soluble protein，and anthocyanin in the flesh，as well as Vc and edible rate of fruit(P<0.05). In the same amount of organic fertilizer，the contents of soluble solids and Vc and edible rate of fruit with applications of biochar decreased significantly compared with those of non-application，but soluble protein increased(P<0.05). The contents of reducing sugar with applications of biochar significantly decreased compared with that of non-application in the low rate of organic fertilizer(22500 kg/hm²)treatment，but significantly increased in the medium rate of organic fertilizer(45000 kg/hm²)treatment(P<0.05). In the same mixture proportion of biochar，the contents of anthocyanin，soluble solids，and edible rate of fruit significantly decreased with the increase of organic fertilizer(P<0.05). The annual yields of red pitaya fruit was not significantly affected by different treatments(P>0.05)，while the annual yield in applications of 3% biochar combined with the medium rate of organic fertilizer and the high rate of organic fertilizer (90000 kg/hm²)treatments increased compared with non-application. The annual yield of the high rate of organic fertilizer with 3% biochar treatment was 34.18 t/hm²，which was the highest among all treatments. The principal component analysis considering quality and yield revealed that the comprehensive assessment of the low and medium rate of organic fertilizer without biochar and the high rate of organic fertilizer with 3% biochar treatments was better than others. In conclusion，the low and medium rate of organic fertilizer without biochar，and the high rate with 3% biochar is recommended.