Tissue Culture And Regeneration System Establishment Of Amelanchier Alnifolia Nutt

Stems with buds of three-year old Amelanchier alnifolia Nutt were used as a source of initial explants in this study. We studied on the effect of factors which affected the tissue culture, such as pre-sterilization, the basic medium and plant growth regulator during the stage of initiation culture, subculture and rooting. In the research of inducing callus and regeneration for saskatoon's leaves and stems, we studied the suitable basic medium, plant growth regulator and other factors, and also studied the effect of silver nitrate on leaves regeneration. Finally, we gained the strongly regenerated plantlet. The results are as follows: 1.The best method for sterilize of saskatoon's stems was dip in 70% ethanol for 30s ,then agitating in a solution of NaClO (10% active chlorine) for 10 minutes ,followed by washing tow times in sterile distilled water, and then agitated in 0.1% HgCl2 for 2 minutes, and finally rinsed five times with sterile distilled water. 2. The suitable medium for initiation culture of saskatoon's stems was MS with IAA 0.1mg/L , 6-BA 2.0mg/L。3. The best basic medium for multiplication was MS, and the suitable growth hormone was IBA, the suitable cytokinin was 6-BA and its suitable concentration was 2.0mg/L. IAA1.0mg/L with 6-BA 2.0mg/L or IBA 0.1mg/L with 6-BA 2.0mg/L can also greatly accelerate the multiplication. GA3 had no active effect but restraining the plantlets growth. In this period, the sucrose could be substituted for white sugar to reduce the cost. 4. The best medium for rooting was 1/4MS with NAA 0.1mg/L. The quality of the root system was better than others, and the rooting rate was 96.43%. The survive ratio of the first class rooting plantlets could reach 100% when they were transplanted to the medium composed with one part vermiculite and two parts humus. The humidity should be controlled strictly. 5. The best medium for inducing callus was H medium, and the ideal hormones composing was NAA 2.0 mg/L with 6-BA 1.0mg/L. The best explants was leaves. It could be induced to root on the medium of 1/4MS with NAA1.0 mg/L, 1/2MS withIBA0.5 mg/L or MS with NAA6.0mg/L and KT2.0mg/L. The callus cultured on the medium of MS with 6-BA 6.0mg/L could be regenerated strongly, but the regeneration rate should be increased. The AgNO3 had no active effect for callus inducing and regeneration from leaves.