| The purpose of the study was to establish the cryopreservation system of Callus of Robinia Idaho.The result is shown as follows:1.There is some deviation between sample cooling rate and cooling rate of the cold trough by using Controlled rate freezer.This deviation can being reduced by using slower cooling rate and low-temperature stay with appropriate time.The decreasing of deviation Conducive to the improvement of the survival rate of the sample.2.Cryopreservation of callus of Robinia Idaho was carried out by using 4 cryoprotectants.The results showed that the callus treated with cryoprotectants (MS+10%DMSO+0.5mol·L-1 Sucrose) have the highest survival rate.The highest survival rate can reaches 91.81%.(MS+10%DMSO+0.5mol·L-1Sucrose) can being the suitable cryoprotectants for the callus of Robinia Idaho.3.The suitable cryopreservation procedures for callus of Robinia pseudoacacia as follows:the calli were cooled at 4℃for 2h after treated with cryoprotectants (MS+10%DMSO+0.5mol·L-1Sucrose),and further refrigerated at a rate of 1℃·min-1 to-7℃by using controlled rate freezer,Holding for 1h at-7℃,then refrigerated at a rate of 0.1℃·min-1 to-20℃.Holding for lh at-20℃,and further refrigerated at a rate of 0.3℃·min-1 to-40℃.The calli then were preserved in liquid nitrogen after it's temperature reaches-40℃.Cooling rate and low-temperature stay have a significant impact on the vitality of callus of Robinia Idaho.unsuitable cooling mode will significant impacted the follow-up results.4.The thawed calli were washed with liquid MS medium with 6-BA 5 mg·L-1 and 30g·L-1Sucrose,then transferred to solid medium and cultured in darkness.Two weeks later,the calli were cultured in light.The calli of Robinia Idaho showing vigorous growth were fresh and green after 3 days,and the survival rate reached 52%.Calli of Robinia Idaho could regenerate to plantlets.
|
PDF Full Text Request